Network Pharmacology Research Indicates that Wu-Mei-Wan Treats Obesity by Inhibiting Th17 Cell Differentiation and Alleviating Metabolic Inflammation.

BACKGROUND: Wu-Mei-Wan (WMW), a traditional Chinese medicine (TCM) formula, has a good effect on the treatment of obesity and has been proven helpful to promote the metabolism of adipose tissue. However, its underlying mechanism remains to be studied. This study aims to explore the potential pharmacological mechanism of WMW in the treatment of obesity. METHODS: Network pharmacology was used to sort out the relationship between WMW putative targets and obesity-related drug targets or disease targets, which indicated the mechanism of WMW in treating obesity from two aspects of clinical drugs approved by the Food and Drug Administration (FDA) and obesity-related diseases. Databases such as Traditional Chinese Medicine Systems Pharmacology (TCMSP), PubChem, DrugBank, DisGeNET, and Genecards were used to collect information about targets. String platform was used to convert the data into gene symbol of "homo sapiens", and perform gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. With the Human Protein Reference Database (HPRD) as background data, Cytoscape 3.6.0 software was used to construct a new protein-protein interaction (PPI) network. Mechanism diagrams of key pathways were obtained from the KEGG database. AutoDock Vina software was used to conduct molecular docking verification. RESULTS: The number of targets in the overlap between WMW putative targets and obesity-related drug targets accounted for more than 50% of the latter, and HTR3A, SLC6A4, and CYP3A4 were core targets. In obesity-related disease targets-WMW putative targets PPI network, the Th17 cell differentiation pathway, and the IL-17 signaling pathway were key pathways, and the 1st module and the 7th module were central function modules that were highly associated with immunity and inflammation. Molecular docking verified that STAT3, TGFB1, MMP9, AHR, IL1B, and CCL2 were core targets in the treatment of WMW on obesity. CONCLUSION: WMW has similar effects on lipid and drug metabolism as the current obesity-related drugs, and is likely to treat obesity by inhibiting Th17 cell differentiation and alleviating metabolic inflammation.

An analysis on the compatibility principle and efficacy characteristics of Wumei Pill from the perspective of "Tang-Ye-Jing-Fa Map" / 国际中医中药杂志

Wumei Pill is a classic insect repellent prescription, which is used for ascaris lumbricoides and diarrhea. It is considered to be a representative prescription for the treatment of simultaneous occurrence of cold and heat syndromes. Its prescription is mainly composed of sour drugs ( Mume Fructus and bitter wine) and pungent drugs ( Aconiti Lateralis Radix Praeparataaconite, Cinnamomi Ramulus, Sichuan pepper, Angelicae Sinensis Radix, Zingiberis Rhizoma, and Asari Radix et Rhizoma), supplemented by bitter drugs ( Coptidis Rhizoma and Phellodendri Chinensis Cortex) and sweet drugs ( Ginseng Radix et Rhizoma and honey). From the point of view of "Tang-Ye-Jing-Fa Map", the role of liver is to disperse, whose deficiency leads to limb syncope, and excess leads to the full of hypochondrium and abdominal pain. The pungent herbs can tonify the liver-deficiency, the sour herbs can dispel the liver-excess, and sweet herbs can relive them both. The role of spleen is to moderate, whose deficiency leads to fatigue and weakness, and excess leads to the vomiting and diarrhea. The sweet herbs can tonify the spleen-deficiency, the pungent herbs can dispel the spleen-excess, and bitter herbs can relive them both. Therefore, the function of Wumei Pills is located in the liver and spleen, giving consideration to the heart and lung. It is used for the deficiency and excess mixed syndrome of liver and spleen, which is mainly characterized by chest and flank abdominal pain, limb chills, diarrhea, muscle and pulse contracture and the symptoms of the parts along the liver meridian. It is commonly used in the treatment of ascaris, diarrhea, impotence, depression. At the same time, it can also be used for the treatment of diseases about heart and lung, and cough, anxiety and other diseases.

Treatment of ulcerative colitis with Wu-Mei-Wan by inhibiting intestinal inflammatory response and repairing damaged intestinal mucosa.

BACKGROUND: Wu-Mei-Wan (WMW), a traditional Chinese medicine, has been applied in the treatment of gastrointestinal diseases with long-term diarrhea and mucopurulent bloody stool as the main symptoms since ancient times. Studies have shown that WMW inhibits intestinal inflammation, repairs damaged intestinal mucosa, resists colon necrosis, and resists intestinal fibrosis. However, the specific mechanism of action is not yet clear. OBJECTIVE: Ulcerative colitis (UC), an intestinal disease with intestinal inflammation and injury as the main pathological manifestations, is one of the high-risk factors for colon cancer. Inhibiting the inflammatory response and promoting colonic epithelial repair are critical to the treatment of UC. However, there is still a lack of remedies with satisfactory curative effects. In this study, the role of WMW in dextran sulfate sodium (DSS)-induced colitis in mice and its related mechanisms are discussed from two aspects: intestinal inflammation and tissue repair. METHODS: DSS was used to induce colitis in mice and the therapeutic effect of WMW was analyzed by disease activity score, histopathological score, colon length measurement, serum cytokine detection, and flow cytometry. Macrophage activation and colonic stem cell proliferation were observed by immunohistochemistry. The expression of critical molecules in macrophage activation and colonic stem cell proliferation signaling pathways in colon tissue was detected with immunohistochemistry, immunofluorescence staining, RT-qPCR, and Western blot. RESULTS: WMW could significantly alleviate DSS-induced colitis. We showed that WMW could reduce disease activity, reduce pathological scores, limit weight loss, inhibit colon shortening, inhibit inflammatory factor secretion, attenuate inflammatory response, and promote the repair of damaged colonic epithelium. WMW inhibited the activation of colonic macrophages, and its mechanism might be inhibiting the Notch/NF-κB/NLRP3 pathway; WMW promoted the proliferation of colonic stem cells, and its mechanism was associated with the regulation of the Hippo/YAP signaling pathway. CONCLUSION: The results of this study suggested that WMW could treat UC via a mechanism that inhibited the intestinal inflammatory response and repaired damaged intestinal mucosa.

Traditional herbal formula Wu-Mei-Wan alleviates TNBS-induced colitis in mice by inhibiting necroptosis through increasing RIPK3 O-GlcNAcylation.

BACKGROUND: Accumulating evidence indicated that necroptosis plays an essential role in the pathogenesis of inflammatory bowel disease (IBD). The O-linked ß-N-acetylglucosaminylation (O-GlcNAcylation) of necroptotic signal molecule receptor-interacting serine-threonine kinase 3 (RIPK3) was reported to exert a protective effect in gut inflammation. Our recent study suggested traditional Chinese herbal formula Wu-Mei-Wan (WMW) as an effective prescription in mouse colitis. However, the potential mechanisms are not fully understood. Considering the crucial role of necroptosis in the pathogenesis of IBD, therefore, this study was designed to explain whether the anti-colitis effect of WMW is mediated by modulating necroptosis and its related mechanisms. METHODS: The protective effects of WMW on colitis have been determined by detecting colitis mice body weight, disease activity index (DAI), survival rate and colon length. Colonic inflammation was examined by inflammatory cells infiltration and local cytokines levels. After then, we measured the levels of necroptosis and O-GlcNAcylation. C O-immunoprecipitation experiments were used to address whether elevated O-GlcNAcylation can inhibit necroptotic signal transduction in the treatment of WMW. Finally, the key enzymes in O-GlcNAcylation: O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) were examined and molecular docking analysis was used to determine effective natural compounds in the regulation on OGT and OGA activities. RESULTS: Our results showed that WMW significantly improved mice body weight, survival rate and colon length, decreased DAI in TNBS-induced colitis. WMW obviously alleviated colonic inflammatory responses with reduced macrophages, neutrophils infiltration and local IL-1ß, IL-6, TNF-α and IFN-γ levels. It was found that WMW increased colonic O-GlcNAcylation level and inhibited the activation of RIPK1, RIPK3 and MLKL. Then, further experiments revealed that WMW enhanced OGT activity and suppressed OGA activity, thereby increasing RIPK3 O-GlcNAcylation and inhibiting the binding of RIPK3 and MLKL, which led to the inhibition of necroptosis. Additionally, docking analysis demonstrated that hesperidin, coptisine and ginsenoside Rb1 may exert a major role in the regulation on OGT and OGA activities by WMW. CONCLUSION: Our work demonstrated that WMW can alleviate TNBS-induced colitis in mice by inhibiting necroptosis through increasing RIPK3 O-GlcNAcylation.

Wu-Mei-Wan prevents high-fat diet-induced obesity by reducing white adipose tissue and enhancing brown adipose tissue function.

BACKGROUND: Wu-Mei-Wan, a classic traditional Chinese herb medicine, is one of the most important formulations to treat digestive diseases from ancient times to the present. Our previous study showed that WMW treatment can prevent T2DM in db/db mice, which motivating the application of WMW on metabolic disorders. PURPOSE: Obesity and its comorbid diseases have increased dramatically and are now a worldwide health problem. There is still a lack of satisfactory treatment strategies for obesity. This work was designed to assess the effect and related mechanism of WMW on high fat diet (HFD)-induced obese mice model. METHODS: Obese mice were induced by HFD. Thetherapeutic effect of WMW were analyzed by examining body and adipose tissue weight, metabolic profile and energy expenditure. Adipose tissue phenotype was determined by histological staining and the mitochondrial content was examined by transmission electron microscopy (TEM). Immunohistochemical and immunofluorescence staining, RT-qPCR and Western blot analysis were used to evaluate expression of key molecules in adipose tissue. RESULTS: WMW treatment significantly protects HFD-induced obesity. Here we showed that WMW limits weight gain, improves metabolic profile and increases energy expenditure. WMW inhibits the hypertrophy and hyperplasia of white adipocytes, the mechanism involving the inhibition of TLR3/IL-6/JAK1/STAT3 pathway. In brown adipose tissue (BAT), WMW promotes thermogenicprogramme without affecting cell proliferation. The activated BMP7/ Smad1/5/9 pathway is considered to be one of the explanations for the effect of WMW on BAT. CONCLUSION: Our results suggested that WMW can prevent obesity and its underlying mechanisms are associated with reducing white adipose tissue and enhancing brown adipose tissue function.

Wu-Mei-Wan ameliorates chronic colitis-associated intestinal fibrosis through inhibiting fibroblast activation.

ETHNOPHARMACOLOGICAL RELEVANCE: Wu-Mei-Wan (WMW), a classic traditional Chinese herb medicine, is one of the most important formulations to treat digestive diseases from ancient times to the present. Previous study showed that WMW has satisfactory curative effects on experimental colitis, which motivating the application of WMW on colitis-associated complications. AIM OF THE STUDY: Intestinal fibrosis is usually considered to be a common complication of inflammatory bowel disease (IBD), particularly Crohn's disease (CD). Currently, no effective preventive measures or medical therapies are available for that. This work was designed to evaluate the effect and related mechanism of WMW on chronic colitis-associated intestinal fibrosis mice model. MATERIALS AND METHODS: The chronic colitis-associated intestinal fibrosis mice model was established by weekly intrarectal injection of 2,4,6-trinitrobenzene sulfonic acid (TNBS). The mice survival rate, disease activity index (DAI), colon length and histological score were examined to assess the therapeutic effect of WMW. Masson's trichrome staining, hydroxyproline assay, immunohistochemical staining and western blot analysis were used to evaluate fibrosis level. Colon inflammation was determined by ELISA and immunofluorescence staining. Immunofluorescence staining was used to evaluate fibroblasts proliferation and epithelial to mesenchymal transition (EMT), and the expression of key molecules in fibrosis was analyzed by western blot. RESULTS: Here we showed that WMW alleviates chronic colitis with improved survival rate, DAI, colon length and histological score. WMW inhibited the progression of intestinal fibrosis, decreased the expression of various fibrosis markers, such as α-SMA, collagen I, MMP-9 and fibronectin. In addition, WMW treatment reduced cytokines IL-6 and IFN-γ, and downregulated proinflammatory NF-κBp65 and STAT3 signaling pathways. Importantly, administration of WMW led to the inhibition of colon fibroblast proliferation and EMT, which are important mediators during fibrosis. Several key profibrotic pathways, including TGF-ß/Smad and Wnt/ß-catenin pathways, were downregulated by WMW treatment. CONCLUSION: Our work demonstrated that WMW can prevent intestinal fibrosis and the mechanisms involved may be related to the inhibition of colon fibroblasts activation.

Wu-Mei-wan protects pancreatic ß cells by inhibiting NLRP3 Inflammasome activation in diabetic mice.

BACKGROUND: Wu-Mei-Wan (WMW) is a traditional Chinese herbal formulation that is clinically prescribed to treat diabetes mellitus in China. WMW has been shown to alleviate damage in pancreatic ß cells, but the underlying mechanism remains unclear. This study aims to explore how WMW plays a protective role in pancreatic islets. METHODS: Drug testing and mechanism analyses were performed on mice treated with three concentrations of WMW (4800, 9600, and 19,200 mg/kg/bw) for four consecutive weeks. Blood was collected from both db/db and wild-type mice to determine fasting blood glucose (FBG) and serum insulin levels. The expression of proteins related to apoptosis, cysteinyl aspartate-specific proteinase 12 (caspase-12) and B-cell leukemia 2 (Bcl-2), was measured by western blot. Interleukin-1ß (IL-1ß), interleukin-18 (IL-18), monocyte chemoattractant protein-1α (MCP-1α), and tumor necrosis factor-α (TNF-α) in the pancreas were tested with enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry staining of F4/80 was performed to measure the pancreatic infiltration of macrophages. Western blot and immunofluorescence staining of the NLRP3 inflammasome were used to measure the expression of proteins related to apoptosis and inflammation. RESULTS: WMW dose-dependently reduced FBG and promoted serum insulin secretion in db/db mice compared to the wild-type controls. WMW protected pancreatic ß cells with a pattern of decreasing caspase-12 and increasing Bcl-2 expression. WMW also reversed the upregulated production of IL-1ß, IL-18, MCP-1α, and macrophage-specific surface glycoprotein F4/80 in diabetic mice. In addition, the protein expression levels of NLRP3 inflammasome components NLRP3, ASC, and caspase-1 (P20) were higher in db/db mice than in wild-type controls. CONCLUSIONS: WMW inhibits the activation of the NLRP3 inflammasome to protect pancreatic ß cells and prevent type 2 diabetes mellitus development.

Clinical observation of Wumeiwan combined with triple therapy for treatment of chronic atrophic gastritis / 国际中医中药杂志

Objective@#To explore clinical efficacy and safety of Wumeiwan combined with triple therapy for treatment of chronic atrophic gastritis (CAG).@*Methods@#According to the random indicator method, 113 patients with CAG were divided into control group (n=56) and treatment group (n=57). Patients of control group were treated with triple therapy, while treatment groupwere treated Wumeiwan combined with triple therapy. The two groups were treated for 3 months. Clinical effect was evaluated after treatment. The helicobacter pylori (Hp) conversion to negative of the two groups was compared and recorded. The Hp overcast conditions of the two groups were compared. The serum TNF-α, IL-6, IL-8 and peripheral blood CD3+, CD4+, CD8+, CD4+/CD8+ of the two groups before and after treatment were compared. The adverse reactions of the two groups during the treatment were compared.@*Results@#Total effective rate of treatment group was 94.7% (54/57), which was significantly higher than the control group 82.14% (46/56), and the difference was statistically significant (χ2=4.401, P=0.036). Hp overcast rate of treatment group was 88.2% (45/51), which was significantly higher than the control group 48.0%(24/50), and the difference was statistically significant (χ2=18.883, P=0.000). After treatment, the serum TNF-α (1.43 ± 0.17 mg/L vs. 1.97 ± 0.22 mg/L, t=14.615), IL-6 (30.79 ± 3.65 ng/L vs. 41.13 ± 4.10 ng/L, t=14.166), IL-8 (7.52 ± 1.32 ng/L vs. 9.60 ± 1.77 ng/L, t=7.090) in the treatment group were lower than those in the control group (P<0.05). After treatment, the peripheral blood CD3+ (75.12% ± 16.44% vs. 67.33% ± 14.37%, t=2.680), CD4+ (39.02% ± 11.41% vs. 33.49% ± 10.61%, t=2.667), CD4+/CD8+ (1.58 ± 0.35 vs. 1.19 ± 0.32, t=6.179) in the treatment group were higher than those in the control group (P<0.05), the peripheral blood CD8+ (24.75% ± 9.69% vs. 28.12% ± 11.29%, t=1.704) in the treatment group were higher than those in the control group (P<0.05). There was no significantly difference of the adverse reaction rates of the two groups during treatment (χ2=0.134, P=0.714).@*Conclusions@#The Wumeiwan combined with triple therapy for treatment of CAG has a good efficacy and low adverse reactions, has serious anti-inflammatory effects, can improve the body immunity, and it was worthy clinical application.

The clinical observation of the repletion and depletion acupuncture combined with Wumei pill for the sleep disorders after stroke / 国际中医中药杂志

Objective In order to evaluate the clinical curative effect of double repletion and depletion acupuncture combined with Wumei pill for the sleep disorders after stroke.Methods A total of 150 patients of sleep disorders after stroke that accorded with the inclusion criteria was randomly divided into the treatment group and the control group.Thirteen patients were off loss in the study with 8.7％.And a total of 137 patients completed the protocol,including 70 in the treatment group and 67 in the control group.Both groups were treated with basic treatment.Besides,the treatment group added the treatment of double repletion and depletion acupuncture combined with Wumei pill;while the control group Estazolam tablet.There were 2 weeks for one course and 2 courses in total with 6 months follow-up.The PSQI,neurokinetic energy assessment and cognitive impairment were evaluated after treatment and 6 months follow-up.After treatment,plasma 5-HT and BDNF level were detected and clinical curative effect was observed.Results The PSQI score after treatment (6.3 ± 0.9 vs.9.3 ± 1.1,t=-3.424) 6 months followed up (4.2 ± 0.3 vs.7.5 ± 0.4,t=-3.675) in the treatment group was significantly lower than those in the control group (P＜0.05).The NIHSS score after treatment (8.2 ± 1.0 vs.11.4 ± 1.0,t=-4.557) and 6 months followed up (5.1 ± 0.5 vs.7.6 ± 0.9,t=-3.135) in treatment group was significantly lower than the control group (P＜0.05).The MMSE score after treatment (22.7 ± 2.9 vs.18.3 ± 2.8,t=-4.691) and 6 months followed up (25.0 ± 2.6 vs.21.5 ± 2.6,t=-4.902) in treatment group were significantly higher than the control group (P＜0.05).After treatment,the level of 5-HT (131.85 ± 11.72 ng/ml vs.101.21 ± 10.23 ng/ml,t=0.014) and BDNF (45.17 ± 6.03 ng/ml vs.38.46 ± 5.28 ng/ml,t=5.406) in treatment group were significantly higher than those in the control group (P＜0.05).After treatment,the treatment group effective rate was 92.9％ (65/70) and 81.6％ (54/67) in the control group.The difference was statistically significant (Z=-2.644,P=0.008).Conclusions The double repletion and depletion of acupuncture combined with Wumei pill can improve the sleep quality and cognitive behavior of patients with post-stroke sleep disorders.The possible mechanism is related with improvement of the expression level of 5-HT and BDNF.