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Back to Journal »Journal of Inflammation Research» Volume 14

In vitro anti-inflammatory effects of four miraculous mushroom water extracts containing psilocybin on 15-lipoxygenase activity and lipopolysaccharide-induced cyclooxygenase-2 and inflammatory cytokines in human U937 macrophages

Author Nkadimeng SM, Steinmann CM, Eloff JN

Published on August 5, 2021 2021 Volume: 14 pages 3729-3738

DOI https://doi.org/10.2147/JIR.S317182

Single anonymous peer review

Reviewing editor: Professor Quan Ning

Sanah Malomile Nkadimeng,1 Christiaan ML Steinmann,2 Jacobus N Eloff1 1Plant Medicine Program, Department of Clinical Sciences, University of Pretoria, Onderstepoort, Pretoria, Gauteng, 0110, South Africa; 2Sefako Makgatho, University of Health Sciences Department, Ga-Rankuwa, Gauteng, 0208, South Africa Tel 27 12 529 8187 Email [email protected] Purpose: During pathological inflammation, macrophages are activated to produce accumulation of inflammatory mediators, such as inducing cyclooxygenase 2 (COX-2), 15-lipoxygenase (15-LOX) and pro-inflammatory cytokines. Pathological inflammation is an important problem for many chronic diseases. Therefore, it is important to conduct more research on natural treatments with anti-inflammatory potential. Since ancient times, mushrooms containing psilocybin, also known as magic mushrooms, have been used to heal the soul and improve the quality of life. However, little is known about its anti-inflammatory potential. This study aims to investigate the anti-inflammatory effects and lipids of four psilocybin-containing mushrooms from Panaeolus and Psilocybe (Panaeolus cyanescens, Psilocybe natalensis, Psilocybe cubensis and Psilocybe cubensis leucistic A strains) against 15-LOX activity in vitro for the first time. Polysaccharide (LPS)-induced inflammation in human U937 macrophages. Method: Mushrooms are grown and extracted with boiling water. The effects of four water extracts on the activity of 15-LOX were determined. Confluent human U937 cells were differentiated with phorbol 12-myristate 13-acetate and treated with hot water extracts (25 and 50 μg/mL) for 2 hours, and then stimulated with 1 μg/mL LPS for 24 hours. Quercetin was used as a positive control. The control cells were differentiated but not induced or treated by LPS. Measure the concentration of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 and IL-10. The levels of COX-2 and mitochondrial activity were also measured. Results: The four aqueous extracts had poor inhibitory activity against 15-LOX, with IC50> 250 μg/mL. The extract is safe at the concentration studied, and significantly inhibits the production of LPS-induced pro-inflammatory mediators, TNF-α and IL-1β, and reduces IL-6 and IL-6 and IL-1β in treated human U937 macrophages. The concentration of COX-2. The water extract also non-significantly increased the survival rate of the treated cells and the level of anti-inflammatory IL-10. Conclusion: Studies have shown that the hot water extracts of four miraculous mushrooms containing psilocybin have potential anti-inflammatory effects, which are achieved by down-regulating pro-inflammatory mediators. Key words: blue spot fish, Usnea, Usnea, Usnea A strain, Pro-inflammatory cytokines, COX-2, 15-LOX

Pathological inflammation is at the core of many chronic diseases, including cardiovascular, cancer, arthritis, neurodegenerative diseases, chronic depression, and aging. 1,2 Inflammation is the natural basic response of our innate immune system when exposed to external factors or injuries, and its purpose is 3 In summary, immunity not only distinguishes self from non-self, but also distinguishes dangerous and non-dangerous factors. 4 However, if inflammation is not properly regulated and prolonged, it will lead to an unhealthy state because it is associated with many chronic diseases. Therefore, during pathological inflammation, the macrophages found in the body are activated, producing unhealthy accumulation of inflammatory mediators. 5

Cytokines are small proteins released by cells that can act on the cell that secretes them, neighboring cells, or in some cases, distant cells. 6 There are pro-inflammatory and anti-inflammatory cytokines. Although produced by many different cell populations, cytokines are mainly produced by helper T cells and macrophages. 6 In particular, pro-inflammatory cytokines are mainly produced by activated macrophages and are involved in the up-regulation of inflammatory responses. 6 The production of inflammatory cytokines such as interleukin (IL) is unbalanced)-6, tumor necrosis factor-α (TNF-α), IL-1 and IL-10 have been found to be related to immune dysfunction and mediate tissue and organ damage Related to inflammation. 7 There is also a lot of evidence that pro-IL-1β, IL-6 and TNF-α and other inflammatory cytokines participate in the process of pathological pain by directly activating nociceptive neurons. 6 Studies have shown that in the process of cell damage, IL-1β cytokines are mainly released by monocytes and macrophages and non-immune cells (such as fibroblasts and endothelial cells) during foreign invasion, infection and inflammation. 6 In addition, IL-1β cytokines have also been found to produce substance P and prostaglandin endoperoxidase 2 (PGE2) in a large number of neurons and glial cells. 6 It was found that IL-1 receptor antagonist is a specific IL-1 receptor antagonist by using IL-a and other drugs to inhibit IL-1β receptor. Cytokine-mediated inflammatory hyperalgesia and allodynia induced by nerve damage. 6 TNF-α is also a major pro-inflammatory cytokine. It acts on various signaling pathways by combining its two cell surface receptors, TNF receptor 1 and TNF receptor 2, to regulate apoptosis pathways and inflammatory NF-kB. Activate and activate stress-activated protein kinases. 6 In some studies, blocking TNF-α resulted in the complete elimination of hyperalgesia, which is usually observed after the administration of lipopolysaccharide (LPS). 6 The cytokine IL-6 has also been found to play a key role in the response of neurons to nerve injury, so its inhibition has led to a decrease in regeneration in some studies. 6

On the other hand, interleukin 10 is considered to be an important anti-inflammatory modulator and has been found to have a preventive effect in pathological inflammation-mediated diseases. 8

One of the well-known metabolic precursors is arachidonic acid (AA), which are involved in many inflammatory pathways. 9 Arachidonic acid is an unsaturated fatty acid with 20 carbons. In the resting state, they are usually distributed in the lipid bilayer membrane. 10 However, various studies have shown that many external and internal factors may stimulate phospholipase A2, the latter After stimulation, the membrane-bound AA is cleaved from the phospholipids, making it available for the three main inflammatory pathways, including cytochrome P-450 monooxygenase, lipoxygenase (LOX) and cyclooxygenase (COX) ) Access. 11 Cyclooxygenase and LOX are the most studied and key enzymes that lead to the progression of inflammation. 11

The COX pathway begins with the formation of hydroperoxide endoperoxide (PGG2) from AA, which will cause oxygen molecules to be added to the AA structure. 12 Then PGG2 binds to a specific location of PGG2, where it is reduced by lipid peroxidation and leads to the production of prostaglandin H2 (PGH2), which is the main metabolic substrate of prostaglandin and thromboxane-related synthase. 13,14 COX enzyme is characterized as two different isoenzymes, namely COX-1, which is constitutive in nature, and COX-2 is mainly expressed in different parts, and it is easily stimulated by different endogenous and exogenous Stimuli include pro-inflammatory cytokines IL-1, IL-6, TNF-α, lipopolysaccharide (LPS) and stress. 11 Once COX-2 is induced, PGH2 overproduces prostaglandin endoperoxide synthase 2 (PGE2) (the main metabolite of COX-2) through prostaglandin E synthase (PGES). PGE2, together with other prostaglandins, lowers the pain threshold and stimulates pain, sensitizes the nerve endings of the central and peripheral nervous system, increases vascular permeability, and creates a pathway for inflammation-related diseases or diseases. 11,15 As a result, inhibiting the effect of COX-2 enzyme will significantly improve the treatment of pathological inflammation.

On the other hand, lipoxygenase is a dioxygenase that does not contain heme iron and is known to catalyze the stereospecific peroxidation of polyunsaturated fatty acids (such as linoleic acid and AA) to the corresponding hydroperoxy derivatives. 16 Lipoxygenase has multiple physiological functions in epithelium, tumors and immune cells, including skin diseases, tumorigenesis and inflammation. 17 These enzymes are widely found in animals, plants and fungi, as well as in the human body, and are involved in the synthesis of prostaglandins and prostaglandins. Leukotrienes also play an important role in stimulating inflammation. 18 LOX enzymes are involved in the development of diseases, and inhibiting them is considered a key step in disease prevention. 19 In particular, the role of 15-LOX has been involved in various inflammation-related diseases. 17 The pro-inflammatory effects of 15-LOX and its metabolite 15(S)-HETE have been shown to include the study of Namgaladze et al. 20, in which the expression of pro-inflammatory cytokines (such as IL-6 and IL-12) induced by LPS was 15-LOX inhibition in macrophages is reduced.

Reactive oxygen species (ROS) are classified as partially reduced oxygen metabolites, which have strong oxidizing ability and are also key signaling molecules that play an important role in the progression of inflammation. 21 ROS is a byproduct of cell metabolism produced by electrons. The transport chain in the mitochondria and the ROS through cytochrome P450.21 are both signaling molecules and mediators of inflammation. They are harmful to cells at high levels or improperly regulated. 21 They oxidize protein and lipid cellular components and damage DNA, thus causing cell damage and death. 21 Chronic or prolonged ROS production is at the core of the progression of inflammatory diseases. 22 It is known that excessive ROS can also cause inflammation by stimulating the release of cytokines and subsequent activation of COX and LOX signaling.17

Numerous studies have shown that, over time, prolonged inflammation may actually cause damage to the tissues around the affected area. 3 Non-steroidal anti-inflammatory drugs (NSAIDs) that act by inhibiting COX-2 activation, such as aspirin, are currently available. 5 However, studies have also reported adverse reactions related to their use, including upper gastrointestinal system disorders and heart burns . 5 Therefore, more research on anti-inflammatory drugs is essential. Natural products with medicinal properties have been used to treat various inflammations. Studies have shown that natural products have structural diversity superior to synthetic compounds, which makes them a potential source of new compounds with potent anti-inflammatory activity. 11

Mushrooms containing siloxibin, often called magic mushrooms, have been used to heal the mind and improve the quality of life since ancient times. Mushrooms are considered safe, with a lethal concentration of 17 kg/70 kg to humans. 23 The fatality rate of mushrooms is rare, and it is mainly reported in combination with other drugs. 23 Mushrooms containing siloxibin are mainly based on their anti-inhibitory effects, and little is known about their anti-inflammatory potential. We have previously reported on the in vitro antioxidant and anti-inflammatory effects of Psilocybe natalensis, a famous magic mushroom RAW 267.4 macrophages grown in South Africa. 24 This study aims to further study the anti-inflammatory effects of four psilocybins-for the first time in vitro 15-lipoxylates from Panaeolus and Psilocybe mushrooms (Panaeolus cyanescens, Psilocybe natalensis, Psilocybe cubensis and Psilocybe cubensis leucistic A strains). Synthase activity and lipopolysaccharide (LPS)-induced inflammation in human macrophages were studied in vitro. The human U937 macrophage cell line has been widely used and classified as a model of the response of mammalian cells to various inflammatory stimuli. 25 We used hot water extracts of four kinds of mushrooms, which is one of the common eating methods used by magic mushroom users.

The protocol of this study was submitted and approved by the Research Ethics Committee of the University of Pretoria, and the protocol number is REC045-18. In South Africa, psilocybin is a Schedule 7 substance and has been approved by the Medical Control Committee (MCC) of the Ministry of Health of South Africa, and the project has been granted a license POS 223/2019/2020.

The spore print of Panaeolus (Copelandia) cyanescens (Pan cyanescens) is commonly known as "Natal Blue Meanie", Psilocybe natalensis (P. natalensis) is commonly known as "Natal Super strength", Psilocybe cubensis (P. cubensis) is commonly known as "Golden teacher" and Psilocybe A cubensis The strains (P. A strains) were respectively designated by SKU numbers TBMN-1, TEA-1, NSS-1 and AAP-1 and the Growth Sterile Substrate Kit (SSK-2) and were purchased from the Spore Spot Company in Durban, South Africa. Upon arrival, the spores were inoculated, grown and harvested according to the method of Nkadimeng et al., and extracted with boiling water. 24 The extract is stored in the refrigerator, protected from light, until use.

According to the method of Ondua et al. 26, based on the formation of composite Fe 3 /xylenol orange (Sigma-Aldrich), there is absorption at 560 nm. The percent inhibition of 15-lipoxygenase activity was determined, and the inhibition concentration (IC50) was determined using a nonlinear regression curve of 15-LOX percent inhibition versus logarithm of the test concentration.

The human U937 macrophage cell line was obtained from ATCC. The cells were grown in RPMI 1640 (Sigma-Aldrich) medium supplemented with 10% fetal bovine serum and 1% penicillin (100 units/mL) and streptomycin (100 units/mL). µg/mL) (Celtic Molecular Diagnostics) at 37 °C and 5% CO2. Once the cell confluence reaches about 80-90%, the cells are harvested. For all experiments, according to the method of Harikrishnan et al., by adding (200 nM) phorbol 12-myristate 13-acetate (PMA) (Biocom Africa) for 24 hours, U937 cells were differentiated to obtain macrophages Phenotype. 5 After 24 hours, wash the cells once with normal medium, and then incubate with serum-free medium for 24 hours for the recovery phase.

Then the cells were modified according to the method of Harikrishnan et al. 5. In short, the differentiated cells were seeded in a 25 cm2 flask (Lasec) at a density of 2×106, and then hot water extracts (25 and 50 µg/mL) and quercetin (50 µg/mL) were used ( Sigma-Aldrich), a well-known antioxidant and flavonol found in many fruits and plants, 2 hours before induction with LPS (1 µg/mL) (Sigma-Aldrich), and at 37°C Culture in a 5% CO2 incubator. LPS cells are differentiated, induced with LPS but not treated. The control cells were differentiated but neither induced nor treated with LPS. Expose and process the cells for more than 24 hours. After 24 hours, remove the medium and store it in a refrigerator at -80°C until the day of analysis.

According to the manufacturer’s manual protocol, the human PTGS2/COX-2 prostaglandin endoperoxidase 2 (PGE2) ELISA kit (Elabscience, Biocom Africa) was used to determine the effect of the extract on COX-2 levels. The absorbance is directly proportional to the concentration of PTGS2/COX-2 in the sample medium. The concentration of human COX-2 in the cell culture medium sample was calculated from the standard curve.

Use human ELISA kits E-EL-H0109, E-EL-H0149, E-EL-H0102 and E-EL- to determine and quantify the effect of the extract on the concentration of TNF-α, IL1 β, IL6 and IL10 H0103, (Elabscience , Biocom Africa) respectively follow the instructions for use of cell culture media. Calculate the cytokine concentration from the standard curve.

The differentiated human U937 macrophages were seeded into a 24-well plate (NEST, Whitehead Scientific) at a density of 5×105 cells/well, and cultured at 37°C in 5% CO2 for 24 hours. The cells were pretreated with four hot water extracts (25 and 50 µg/mL) and a positive control, quercetin (50 µg/mL) for 2 hours, and then induced with (1 µg/mL) LPS for more than 24 hours. just like before. After 24 hours of incubation, the mitochondrial activity was measured using Resazurin Assay Kit AR002 (R & D, Whitehead Scientific) according to the manufacturer's manual. Use the following formula to calculate the percentage of cell viability:% viability = ((sample absorbance/control absorbance)×100). The experiment was repeated 3 times on different occasions.

The results are expressed as the mean ± standard deviation, and statistically significant values ​​are compared using the one-way analysis of variance and the paired multiple comparison program using the Holm-Sidak method. Use Shapiro-Wilk for normality test and Brown-Forsythe for equal variance test. A p-value ≤ 0.050 is considered statistically significant.

Compared with the positive control quercetin, the 15-LOX inhibitory activity of the four hot water extracts is very low, with IC50> 250 µg/mL and IC50 of 12.35 µg/mL, as shown in Table 1. Table 1 The IC50 of four hot water extracts inhibited 15-LOX activity

Table 1 The IC50 of four hot water extracts for inhibiting 15-LOX activity

Compared with unstimulated control cells, stimulation of human U937 macrophages with LPS significantly increased the concentration of COX-2 (p = 0.009), Figure 1. Treatment with four hot water extracts reduced COX-2 and the effect was significant (p = 0.039), with the lowest concentration of PA strain extract (25 µg/mL), Figure 1. Figure 1 The effect of Pan hot water extract (25 and 50 µg/mL) cyanescens, P.natalensis, P.cubensis and PA strain mushrooms and positive control; Quercetin (50 µg/mL) induces LPS within 24 hours The influence of COX-2 concentration. The control cells were differentiated but neither induced nor treated with LPS. LPS: differentiated and LPS induced. (*Statistically significant).

Figure 1 The effect of Pan cyanescens, P. natalensis, P. cubensis and P. A strains and the hot water extracts (25 and 50 µg/mL) of the positive control; Quercetin (50 µg/mL) is effective in 24 hours The effect of COX-2 concentration induced by LPS. The control cells were differentiated but neither induced nor treated with LPS. LPS: differentiated and LPS induced. (*Statistically significant).

Compared with control cells, LPS significantly increased (p = 0.003) the level of TNF-α cytokine production in stimulated cells, Figure 2. The positive control quercetin significantly reversed this effect (p <0.001). The four mushroom extracts can also significantly inhibit the TNF-α concentration at 25 and 50 µg/mL, Figure 2. Figure 2 Inhibition of Pan cyanescens, P. natalensis hot water extract (25 and 50 µg/mL), P. cubensis and P. A strain mushrooms and positive control, quercetin (50 µg/mL) induces LPS TNF-α is produced for more than 24 hours. The control cells were differentiated but neither induced nor treated with LPS. LPS: differentiated and LPS induced. (*Statistically significant).

Figure 2 The inhibitory effect of Pan cyanescens, P. natalensis, P. cubensis and P. A strains hot water extracts (25 and 50 µg/mL) and positive control quercetin (50 µg/mL) on LPS-induction of TNF -α is generated for more than 24 hours. The control cells were differentiated but neither induced nor treated with LPS. LPS: differentiated and LPS induced. (*Statistically significant).

Compared with the control, LPS stimulation significantly increased the production of IL-1β (p <0.001), Figure 3. The four hot water extracts inhibited LPS-induced IL-1β to a level very close to that of the positive control. Figure 3. Compared with the control, LPS also significantly increased the production of IL-6 (p = 0.010), while quercetin Supplement significantly reversed this effect (p = 0.003), Figure 3. The four hot water extracts reduced the production of IL-6 in the treated cells however; only P. natalensis at concentrations of 25 and 50 µg/mL (p = 0.025 and p = 0.015, respectively) and the highest concentration of 50 µg/mL The inhibitory effect is significant when P. cubensis (p = 0.040), Figure 3. Figure 3 Pan cyanescens, P. natalensis, P. cubensis and P. A strains hot water extracts (25 and 50 µg/mL) and quercetin inhibit LPS-induced pro-inflammatory IL-1β and IL production -6 Cytokines in the 24-hour treatment period. The control cells were differentiated but neither induced nor treated with LPS. LPS: differentiated and LPS induced. (*Statistically significant).

Figure 3 Pan cyanescens, P. natalensis, P. cubensis and P. A strains hot water extracts (25 and 50 µg/mL) and quercetin inhibit LPS-induced pro-inflammatory IL-1β production and IL -6 Cytokines in the 24-hour treatment period. The control cells were differentiated but neither induced nor treated with LPS. LPS: differentiated and LPS induced. (*Statistically significant).

Compared with the control, LPS stimulation non-significantly reduced the IL-10 concentration of induced cells, Figure 4. The four water extracts non-significantly increased the LPS-induced IL-10 production of the treated cells, the same as quercetin. The effect was more obvious when using P. cubensis mushrooms, Figure 4. Figure 4 Pan cyanescens, P. natalensis, P. cubensis and P. A hot water extracts (25 and 50 µg/mL) effect of quercetin on anti-inflammatory IL-10 cytokine levels during 24 hours of treatment Influence. LPS: differentiated and LPS induced.

Figure 4 Pan cyanescens, P. natalensis, P. cubensis and P. A strains hot water extracts (25 and 50 µg/mL) and quercetin anti-inflammatory IL-10 cytokine levels over 24-h treatment Expect. LPS: differentiated and LPS induced.

After treating the LPS-induced cells with the four extracts, the percentage of cell survival increased by more than 100%, similar to the positive control quercetin, Figure 5. Pan cyanescens, P.cubensis and P.natalensis have the highest percentage of survival rate, while the percentage of cell survival rate is lower in concentration and shows a dose-dependent pattern of deceleration. The percentage of cell viability decreases with increasing concentration, while the PA strain increases with increasing concentration. The percentage of vitality is the same as that of quercetin, Figure 5. Figure 5 The effect of Pan cyanescens, P. natalensis, P. cubensis and P. A strain mushroom extract and quercetin on the percentage of cell viability of human U937 macrophages induced by LPS during 24 hours of treatment.

Figure 5 The effect of Pan cyanescens, P. natalensis, P. cubensis and P. A strain mushroom extracts and quercetin on the percentage of cell viability of human U937 macrophages induced by LPS during 24 hours of treatment.

Although the magic mushroom containing psilocybin has been used for centuries for its spiritual healing properties, little is known about its anti-inflammatory potential. Many studies have shown that pathological inflammation is the center of many chronic diseases, including cardiovascular, cancer, arthritis, neurodegeneration, chronic depression, and aging. 2,27,28 Many studies have also shown that long-term inflammation may cause harm over time. 3 In addition, studies have also reported the use of current non-steroidal anti-inflammatory drugs to treat inflammatory diseases (including upper gastrointestinal disorders). Side effects, so more research on treatments (especially natural products) with anti-inflammatory properties is needed. 5,11 The purpose of this study is to study the anti-inflammatory effects of four well-known magic mushrooms containing psilocybin, Pan cyanescens, P. natalensis, P. cubensis and P. Strain A, for the first time in vitro studies on 15-LOX activity and LPS-induced inflammation of human U937 macrophages to establish their potential use in inflammatory diseases. We used the hot water extraction method, which is one of the most commonly used edible methods for magic mushroom users.

This study examined the potential inhibitory ability of mushroom extracts on 15-LOX activity, which is one of the important enzymes in leukotriene biosynthesis and plays an important role in the pathophysiology of many inflammatory diseases. Our research shows that hot water mushroom extracts of all four mushrooms containing psilocybin have very low inhibitory properties, as shown by high IC50> 250 μg/mL for 15-LOX activity.

We further studied the anti-inflammatory effects of four hot water mushroom extracts on LPS-induced human U937 macrophages. This is a widely used cell line classified as a mammalian cell response model to various inflammatory stimuli . 25 Lipopolysaccharide activates macrophages to produce unhealthy accumulation of inflammatory mediators, such as inducible COX-2 and nitric oxide synthase (iNOS), as well as pro-inflammatory cytokines such as TNF-α, IL-1β and IL-6.5. In our study, LPS stimulation increased the concentration of COX-2. Compared with unstimulated control cells, TNF-α, IL-1β, and IL-6 in stimulated cells increased significantly, which is different from previous studies. Unanimous. 5

The COX-2 effect induced by LPS was significantly reversed by quercetin and P. A mushroom extract at a concentration of 25 μg/mL. Treatment with other mushroom extracts also reduced the COX-2 concentration induced by LPS, even if not significantly. Inducible COX-2 enzymes play a key role in pathological inflammation. They cause edema, pain, and tissue damage associated with the disease process. 29 Therefore, it has been established that therapeutic methods with the ability to inhibit the expression or concentration of COX-2 are used by many studies as a valuable tool for the prevention or treatment of pathological inflammatory diseases. 29

In our study, the treatment of four hot water mushroom extracts reversed LPS-induced TNF-α and IL-1β, and significantly inhibited the concentration of these two key pro-inflammatory cytokines in a dose-dependent manner. , Very close to the positive control, quercetin. Studies have shown that TNF-α and IL-1β are one of the main pro-inflammatory cytokines secreted during the development of chronic inflammatory diseases. Therefore, by inhibiting these two key pro-inflammatory cytokines, these extracts show potentially potent anti-inflammatory properties, which may be beneficial for the treatment of chronic inflammatory diseases. A disease particularly affected by inflammation is osteoarthritis (OA), which affects approximately 25% of the world’s population and is characterized by the destruction of the function and structure of articular cartilage. 30 Studies have shown that the pro-inflammatory cytokine IL-1B is an important key factor for the development and progression of OA by stimulating pro-catabolism and pro-inflammatory chondrocyte (chondrocyte) response. 30 Chondrocytes play an important role in maintaining normal tissue synthesis and renewal of old extracellular matrix. 30 Therefore, the catabolic reaction induced by IL-1β promotes the inhibition of cartilage-related gene expression and increases the expression of collagenase and matrix metalloproteinases. The expression of genes involved in matrix degradation. 30 In addition, IL-1β also stimulates chondrocytes to increase the expression of pro-inflammatory genes. Related proteins and genes, such as iNOS and COX-2, will further reduce the main components of the cartilage extracellular matrix when they are induced. 32 Studies have also shown that inhibiting COX-2 expression has an inhibitory effect on OA. 33 In addition, in the study of Yang et al. 31 Among them, IL-1β is used to induce the hypertrophy of the degenerated chondrocyte model, and the expression of many proteins and enzymes including COX-2, PGE-2 and collagen X are up-regulated and increased. These are all used in the study of the powerful COX- 2 Reversal of inhibitor treatment. 31 As a result, the hot water extracts of the four magic mushrooms can significantly inhibit the induced IL-1β and reduce the induced COX-2 level (the P. A strain is significantly treated), making them a good medicine for use Treat inflammatory diseases such as AO.

In addition, P. natalensis and P. cubensis mushroom extracts significantly inhibited LPS-induced IL-6 concentration, while other mushroom extracts did not significantly reduce the level in human macrophages. We have also observed that in our study, the anti-inflammatory cytokine IL-10 is reduced in human macrophages induced by LPS. IL-10 is an important regulator and has a preventive effect in pathological inflammatory conditions. Kind of mushroom extract and quercetin.

None of these effects are due to cytotoxicity, as indicated by the percentage increase in cell viability of cells treated with all four mushroom extracts by more than 100%.

In summary, this study showed that the pro-inflammatory mediator COX-2, pro-inflammatory cytokines TNF-α, IL-1β and IL-6 significantly increased, and these were significantly reversed by quercetin in human U937 macrophages stimulated by LPS. The hot water extract of P. A strain significantly inhibited COX-2 induced by LPS, while other extracts did not significantly reduce the level. This effect is important because therapies that inhibit COX-2 are considered valuable tools to prevent or treat pathological inflammatory diseases. 29 The study also revealed that the four hot water mushroom extracts of Pan cyanescens, P. natalensis and P. cubensis and P. A strain significantly inhibited the two key pro-inflammatory cytokines TNF-α and IL- in a dose-dependent manner. 1β, and very close to quercetin, which is a well-known potent anti-inflammatory flavanol and antioxidant found in many fruits and plants. In particular, the mushroom treatment observed in the study has an effect on the induced IL-1β The significant inhibitory effect of serotonin and the reduction of COX-2 indicate their potential use, especially in inflammation-related diseases such as AO. It is strongly recommended to further study the potential of these magic mushrooms to reduce OA. In addition, P. natalensis and P. cubensis also significantly inhibited LPS-induced IL-6 in human U937 macrophages, while other hot water extracts reduced the level, although not significantly. The extract also did not significantly increase the concentration of the anti-inflammatory cytokine IL-10 in treated human macrophages. These effects are not caused by toxicity, as indicated by more than 100% increase in cell viability in the treated cells. Previous studies have shown that there are fungal chemical compounds in mushrooms containing psilocybin, such as alkaloids and saponins. They are called strong antioxidants, which can neutralize free radicals and flavonoids with antioxidant and anti-inflammatory activities. , And tannins with antioxidant properties, especially in Pan cyanescens and P. cubensis. 32,33 We also previously reported the presence of well-known anti-inflammatory and antioxidant compounds in P. natalensis mushroom extracts. 24 In addition, our previous research also showed a significant inhibitory effect on Pan cyanescens and P. ROS. 34 Excessive ROS is known to cause inflammation by stimulating the release of cytokines and subsequently activating COX and LOX signaling in pathological inflammatory diseases. 17 The presence of these compounds in mushroom extracts may have played a role in inhibiting the key pro-inflammatory cytokines and the aspiration of COX-2 observed in this study. However, the study showed that the four kinds of hot water mushroom extracts had poor inhibitory effects on 15-LOX activity, indicating that these extracts may not need to induce their anti-inflammatory effects through this pathway.

In summary, this study showed that the hot water extracts of Pan cyanescens, P. natalensis, P. cubensis and P. A strains of magic mushrooms inhibited the inflammatory response induced by LPS stimulation in human U937 macrophages. The study also proposes that these effects are achieved through mechanisms involving the inhibition of pro-inflammatory mediators such as COX-2 and pro-inflammatory cytokines. It is recommended to further study the basic mechanism of in vivo and in vitro effects.

We thank Mr. Llewellyn Morland for assisting in mushroom cultivation, and Ms. Lebogang E Moagi for assisting in statistics.

The research was funded by SM Nkadimeng from the Department of Health and Welfare Education and Training Agency (HWSETA) and MJ Medtech. The funder has no role in the research design; in the collection, analysis, or interpretation of the data; in the writing of the manuscript; or in the decision to publish the results.

The author declares that there is no conflict of interest in this work.

1. Ferrucci L, Fabbri E. Inflammation: chronic inflammation in aging, cardiovascular disease and weakness. Nat Rev Cardiol. 2018;15(9):505–522. doi:10.1038/s41569-018-0064-2

2. Liu Yuzhen, Wang Yuxia, Jiang CL. Inflammation: a common pathway for stress-related diseases. Front Hum Neurosci. 2017; 11:316. doi:10.3389/fnhum.2017.00316

3. Muniandy K, Gothai S, Badran KMH, Suresh Kumar S, Esa NM, Arulselvan P. By inhibiting the NF-κB pathway and inhibiting the LPS-induced RAW 264.7 macrophage pro-inflammatory cytokines and medium. Journal of Immunology. 2018; 2018: 3430684. doi:10.1155/2018/3430684

4. Matzinger PT. Danger, there is also a big family. Annu Rev Immunology. 1994;12(1):991-1045. doi:10.1146/annurev.iy.12.040194.005015

5. The anti-inflammatory effects of Harikrishnan H, Jantan I, Haque MA, Kumolosasi E. Phyllanthus amarus Schum. & Ton. By inhibiting LPS-induced NF-κB, MAPK and PI3K-Akt signaling pathways in human macrophages. BMC Complementary Alternative Medicine. 2018;18(1):224. doi:10.1186/s12906-018-2289-3

6. Zhang JM, An J. Cytokines, inflammation and pain. International Anesthetics Clinic. 2009;45(2):27-37. doi:10.1097/AIA.0b013e318034194e

7. Lauwerys BR, Houssiau FA. Cytokines are involved in the pathogenesis of systemic lupus erythematosus. Adv Exp Med Biol. 2003;520:237-251. doi:10.1007/978-1-4615-0171-8_14

8. Porro C, Gianciulli A, Panaro MA. The regulatory role of IL-10 in neurodegenerative diseases. Biomolecules. 2020;10(7):1017. doi:10.3390/biom10071017

9. Zelding DC. The cyclooxygenase pathway of arachidonic acid metabolism. J Biochemistry. 2001;276(39):36059–36062. doi:10.1074/jbc.R100030200

10. Sevanian A, Kim E. Phospholiphase A2 depends on the release of fatty acids from the peroxide film. Free radical biomedicine. 1985;1(4):263–271. doi:10.1016/0748-5514(85)90130-8

11. Attiq A, Jalil J, Husain K, Ahmad W. Use natural products to fight inflammation. Former pharmacist. 2018; 9:976. doi:10.3389/fphar.2018.00976

12. Samuelsson B, Goldyne M, Granström E, etc. Prostaglandins and thromboxanes. Annu Rev Biochem. 1978;47(1):997-1029. doi:10.1146/annurev.bi.47.070178.005025

13. Sugimoto Y, Inazumi T, Tsuchiya S. The role of prostaglandin receptors in female reproduction. J Biochemistry. 2015;157(2):73-80. doi:10.1093/jb/mvu081

14. Kawahara K, Hohjoh H, Inazumi T, Tsuchiya S, Sugimoto Y. Prostaglandin E 2 Induced Inflammation: Correlation of Prostaglandin E Receptors. Journal of Biochim Biophys. 2015;1851(4):414–421. doi:10.1016/j.bbalip.2014.07.008

15. Mbonye UR, Yuan C, Harris CE, etc. Two different pathways of cyclooxygenase-2 protein degradation. J Biochemistry. 2008;283(13):8611–8623. doi:10.1074/jbc.M710137200

16. Kuhn H, Humeniuk L, Kozlov N, Roigas S, Adel S, Heydeck D. The evolutionary hypothesis of the reaction specificity of the mammalian ALOX15 ortholog. Prog lipid reservoir. 2018; 72: 55-74. doi:10.1016/j.plipres.2018.09.002

17. Singh NK, Rao GN. 12/15-A new role of lipoxygenase (ALOX15) in human pathology. Prog lipid reservoir. 2019; 73: 28-45. doi:10.1016/j.plipres.2018.11.001

18. Lončarić M, Strelec I, Moslavac T, Šubarić D, Pavić V, Molnar M. Inhibition of lipoxygenase by plant extracts. Biomolecules. 2021; 11(2):152. doi:10.3390/biom11020152

19. Srivastava P, Vyas VK, Variya B, etc. Synthesis, anti-inflammatory, analgesic, 5-lipoxygenase (5-LOX) inhibitory activity and molecular docking of 7-substituted coumarin derivatives. Biochemistry 2016; 67: 130-138. doi:10.1016/j.bioorg.2016.06.004

20. Namgaladze D, Snodgrass RG, Angioni C, etc. AMP-activated protein kinase inhibits the expression of arachidonic acid 15-lipoxygenase in human macrophages polarized by interleukin-4. J Biochemistry. 2015;290(40):24484–24494. doi:10.1074/jbc.M115.678243

21. Mittal M, Siddiqui MR, Tran K, Reddy SP, Malik AB. Active oxygen in inflammation and tissue damage. Anti-oxidant redox signal. 2014;20(7):1126–1167. doi:10.1089/ars.2012.5149

22. Griffith B, Pendyala S, Hecker L, etc. NOX enzyme and lung disease. Anti-oxidant redox signal. 2009;11(10):2505-2516. doi:10.1089/ars.2009.2599

23. Amsterdam JGC, Opperhuizen A, van den Brink W. The potential hazards of magic mushrooms: a review. Regul Toxicol Pharmacol. 2011;59(3):423–429. doi:10.1016/j.yrtph.2011.01.006

24. Nkadimeng SM, Nabatanzi A, Steinmann CML, Eloff JN. The phytochemical, cytotoxic, antioxidant and anti-inflammatory effects of Psilocybe natalensis. plant. 2020; 9(9):127. doi:10.3390/plants9091127

25. Grkovich A, Johnson CA, Buczynski MW, Dennis EA. The expression of cyclooxygenase 2 induced by lipopolysaccharide in human U937 macrophages is dependent on phosphatidic acid phosphohydrolase 1. J Biochemistry. 2006;281(44):32978–32987. doi:10.1074/jbc.M605935200

26. Ondua M, Njoya EM, Abdalla MA, McGaw LJ. Anti-inflammatory and antioxidant properties of leaf extracts of 11 South African medicinal plants traditionally used to treat inflammation. J National Pharmaceutical Journal. 2019;234:27-35. doi:10.1016/j.jep.2018.12.030

27. Nabatanzi A, Nkadimeng SM, Lall N, Kabasa JD, McGaw LJ. Antioxidant and anti-inflammatory activity of Kigelia Africana (Lam.) Benth. Evidence-based complement and alternative medicine. 2020; 2020: 1-11. doi:10.1155/2020/4352084

28. Needleman P, Manning PT. The interaction between the inducible cyclooxygenase (COX-2) and nitric oxide synthase (iNOS) pathways: the impact of therapeutic interventions for osteoarthritis. Bone joint cartilage. 1999; 7(4): 367-370. doi:10.1053/joca.1998.0237

29. Ballaf HS. The efficacy of the latest COX-2 selective inhibitors in rheumatic diseases. Curr Pharm Des. 2007;13(22):2228-2236. doi:10.2174/138161207781368819

30. Jia Y, He W, Zhang H, et al. Morusin improves IL-1β-induced chondrocyte inflammation and osteoarthritis through the NF-κB signaling pathway. The drug Devel Ther. 2020; 14:1227-1240. doi:10.2147/DDDT.S244462

31. Yang YR, Yang XF, Duan HC, Qiao Jinqi. The Cyclooxygenase-2 inhibitor rofecoxib prevents chondrocyte hypertrophy through the Wnt/β-catenin pathway. J Biol Regul Homeost Agents. 2020;34(3):785–794.

32. Bustillos RG, Dulay RMR, Bauto JJ, etc. Panaeolus cyanescens mycelium and fruit body have fungal chemical characteristics and their antioxidant properties are the best submerged culture conditions. Int J Pure Appl Biosci. 2014; 2(6): 175–181.

33. Dhanasekaran D, Latha S, Suganya P, etc. Taxonomic identification and characterization of biologically active compounds of Psilocybe cubensis DPT1 to explore its antibacterial and mosquito larval abilities. Microbial pathogens. 2020;143:104138. doi:10.1016/j.micpath.2020.104138

34. Nkadimeng SM, Steinmann CML, Eloff JN. The effect and safety of Psilocybe cubensis and Panaeolus cyanescens magic mushroom extract on endothelin 1 induced cardiomyocyte hypertrophy and cell damage. Scientific Reports 2020; 10(1): 22314. doi:10.1038/s41598-020-79328-5

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