Wumei pills attenuates 5-fluorouracil-induced intestinal mucositis through Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-κB pathway and microbiota regulation.

BACKGROUND: Radiotherapy and chemotherapy can kill tumor cells and improve the survival rate of cancer patients. However, they can also damage normal cells and cause serious intestinal toxicity, leading to gastrointestinal mucositis[1]. Traditional Chinese medicine is effective in improving the side effects of chemotherapy. Wumei pills (WMP) was originally documented in the Treatise on Exogenous Febrile Diseases. It has a significant effect on chronic diarrhea and other gastrointestinal diseases, but it is not clear whether it affects chemotherapy-induced intestinal mucositis (CIM). AIM: To explore the potential mechanism of WMP in the treatment of CIM through experimental research. METHODS: We used an intraperitoneal injection of 5-fluorouracil (5-Fu) to establish a CIM mouse model and an oral gavage of WMP decoction (11325 and 22650 mg/kg) to evaluate the efficacy of WMP in CIM. We evaluated the effect of WMP on CIM by observing the general conditions of the mice (body weight, food intake, spleen weight, diarrhea score, and hematoxylin and eosin stained tissues). The expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1ß, and myeloperoxidase (MPO), as well as the Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor-κB (TLR4/MyD88/NF-κB) signaling pathway proteins and tight junction proteins (zonula occludens-1, claudin-1, E-cadherin, and mucin-2) was determined. Furthermore, intestinal permeability, intestinal flora, and the levels of short-chain fatty acids (SCFA) were also assessed. RESULTS: WMP effectively improved the body weight, spleen weight, food intake, diarrhea score, and inflammatory status of the mice with intestinal mucositis, which preliminarily confirmed the efficacy of WMP in CIM. Further experiments showed that in addition to reducing the levels of TNF-α, IL-1ß, IL-6, and MPO and inhibiting the expression of the TLR4/MyD88/NF-κB pathway proteins, WMP also repaired the integrity of the mucosal barrier of mice, regulated the intestinal flora, and increased the levels of SCFA (such as butyric acid). CONCLUSION: WMP can play a therapeutic role in CIM by alleviating inflammation, restoring the mucosal barrier, and regulating gut microbiota.

Mechanisms of Banxia Xiexin Decoction Underlying Chronic Atrophic Gastritis via Network Pharmacology, Molecular Docking, and Molecular Dynamics Simulations.

Chronic atrophic gastritis (CAG) is a common chronically digestive disease which is notoriously characterized by atrophy of the epithelium and glands of the gastric mucosa, reduced number, thinning of the gastric mucosa, thickening of the mucosal base, or pyloric glandular hyperplasia and intestinal glandular hyperplasia, or with atypical hyperplasia. Banxia Xiexin decoction (BXD) has been applied for two thousand years and is considered an effective therapy for functional dyspepsia, gastroesophageal reflux disease and colon cancer. In this current study, to probe into the underlying mechanism of BXD on CAG, network pharmacology was conducted to collect druggable ingredients and predicted targets of BXD and the CAG-associated targets were harvested to take intersection with druggable ingredients from BXD predicted targets to obtain potential critical action targets. Subsequently, GO enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were conducted to elucidate the underlying mechanisms and roles from the perspective of overall pathways and cellular functions. Eventually, molecular docking integrated with molecular dynamics simulations was conducted to further investigate the mechanism of action of BXD active ingredients on CAG from drug molecule-target interactions and to provide a theoretical basis for BXD drug development.

Effect of Tai Chi on Cognitive Function among Older Adults with Cognitive Impairment: A Systematic Review and Meta-Analysis.

BACKGROUND: Cognitive decline occurs in all persons during the aging process and drugs can only alleviate symptoms and are expensive. Some researches demonstrated that Tai Chi had potential in preventing cognitive decline while others' results showed Tai Chi had no influence on cognitive impairment. Therefore, we conduct a systematic review and meta-analysis to assess the efficacy and safety of cognitive impairment patients practicing Tai Chi. METHODS: A comprehensive literature search was carried out in multiple databases, including PubMed, Cochrane, MEDLINE (Ovid), Web of Science, Embase, Scopus, PsycInfo (Ovid), CKNI, Wan Fang, VIP, SinoMed, and ClinicalTrails, from their inception to 1 July 2020 to collect randomized controlled trials about practicing Tai Chi for patients with cognitive impairment. Primary outcomes included changes of cognitive function and secondary outcomes included changes of memory functions. Data were extracted by two independent individuals and Cochrane Risk of Bias tool version 2.0 was applied for the included studies. Systematic review and meta-analysis were performed by RevMan 5.3 software. RESULTS: The results included 827 cases in 9 studies, of which 375 were in the experimental group and 452 were in the control group. Meta-analysis showed that Mini-Mental State Examination WMD = 1.52, 95% CI [0.90, 2.14]; Montreal Cognitive Assessment WMD = 3.5, 95% CI [0.76, 6.24]; Clinical Dementia Rating WMD = -0.55, 95% CI [-0.80, -0.29]; logical memory delayed recall WMD = 1.1, 95% CI [0.04, 2.16]; digit span forward WMD = 0.53, 95% CI [-0.65, 1.71]; and digit span backward WMD = -0.1, 95% CI [-0.38, 0.19]. No adverse events were reported in the included articles. CONCLUSION: There is limited evidence to support that practicing Tai Chi is effective for older adults with cognitive impairment. Tai Chi seems to be a safe exercise, which can bring better changes in cognitive function score.

Gut Bacteria Selectively Altered by Sennoside A Alleviate Type 2 Diabetes and Obesity Traits.

Accumulating evidences implicate that gut microbiota play an important role in the onset and prolongation of fat inflammation and diabetes. Sennoside A, the main active ingredient of Rhizoma Rhei (rhubarb), is widely used for constipation as a kind of anthranoid laxative (e.g., senna). Here, we put forward the hypothesis that the structural alteration of gut microbiota in obesity mice may be involved in the pathogenesis of type 2 diabetes (T2D) which may be ameliorated by Sennoside A. We investigated the appearance of obesity, insulin resistance, host inflammation, and leaky gut phenotype with or without Sennoside A in db/db mice. Horizontal fecal microbiota transplantation (FMT) was used to confirm the critical roles of gut microbiota in the amelioration of the indices in T2D mice after Sennoside A treatment. As a result, we found that Sennoside A administration markedly improved the indices in T2D mice and obesity-related traits including blood glucose level, body weight, lipid metabolism disorder, and insulin resistance. The gut microbiota changed quickly during the onset of T2D in db/db mice, which confirmed the hypothesis that gut microbiota was involved in the pathogenesis of T2D. Sennoside A altered gut microbial composition which might mediate the antiobesogenic effects in T2D remission. Sennoside A also reduced inflammation and increased tight junction proteins in the ileum in gene-deficient mice via gut microbiota alteration. FMT lowered the blood glucose level and improved insulin resistance, corroborating that Sennoside A perhaps exerted its antiobesogenic effects through gut microbiota alteration. Chemical Compounds Studied in This Article. Compounds studied in this article include Sennoside A (PubChem CID: 73111) and metformin hydrochloride (PubChem CID: 14219).

Xiao-Yao-San Formula Improves Cognitive Ability by Protecting the Hippocampal Neurons in Ovariectomized Rats.

Xiao-Yao-San (XYS) decoction is a traditional Chinese medicine formula. This study aimed to investigate the effect of XYS on cognitive abilities and its underlying mechanism in ovariectomized rats. Female Sprague-Dawley rats were ovariectomized and treated with XYS (3 g/kg or 9 g/kg) by gavage, with subcutaneous injection of 17-ß estradiol (E2, 2 µg/kg) as a positive drug control and gavage of 1 ml saline (0.9%) as a placebo control. After 6 weeks of treatment, rats were examined using the Morris water maze test. The estradiol level in the serum and hippocampus was measured by ELISA. Golgi staining was performed to observe neuronal morphology in the hippocampus. Apoptosis of hippocampal cells was observed by TUNEL staining. The protein content of N-methyl-D-aspartate receptor (NMDAR) 2A and 2B in the hippocampal CA1 region was determined by Western blot and immunohistochemistry. Expression of estrogen receptor (ER) and PI3K signaling was detected by Western blot. Compared with the sham group, both learning and memory were impaired in ovariectomized rats. Rats treated with E2 or high-dose XYS showed better learning and memory compared with the saline-treated rats. High-dose XYS significantly reduced escape latency in the spatial acquisition trial; meanwhile, the cross times and duration in the probe quadrant were increased in the spatial probe trial. High-dose XYS promoted the de novo synthesis of E2 content in the hippocampus but had no significant effect on the serum E2 level. Golgi staining indicated that high-dose XYS could increase the branch number and density of dendritic spines in the hippocampal CA1 area. TUNEL staining showed that high-dose XYS alleviated ovariectomy-induced neuronal apoptosis. The expression level of NMDAR2A and NMDAR2B in hippocampal CA1 was upregulated by XYS treatment. The beneficial effect of XYS was through activating ERα-PI3K signaling. In conclusion, high-dose XYS treatment can improve the cognitive abilities of ovariectomized rats by protecting the hippocampal neurons and restoring the hippocampal E2 level.

Exploring the 'cold/hot' properties of traditional Chinese medicine by cell temperature measurement.

Context: It is common sense that chewing a mint leaf can cause a cooling feeling, while chewing ginger root will produce a burning feeling. In Traditional Chinese Medicine (TCM), this phenomenon is referred to as 'cold/hot' properties of herbs. Herein, it is reported that TCM with different "cold/hot" properties have different effects on the variation of cells.Objective: To explore the intrinsic 'cold/hot' properties of TCM from the perspective of cellular and molecular biology.Materials and methods: A375 cells were selected using Cancer Cell Line Encyclopaedia (CCLE) analysis and western blots. Hypaconitine and baicalin were selected by structural similarity analysis from 56 and 140 compounds, respectively. A wireless thermometry system was used to measure cellular temperature change induced by different compounds. Alteration of intracellular calcium influx was investigated by means of calcium imaging.Results: The IC50 values of GSK1016790A, HC067047, hypaconitine, and baicalin for A375 cells are 8.363 nM, 816.4 µM, 286.4 µM and 29.84 µM, respectively. And, 8 µM hypaconitine induced obvious calcium influx while 8 µM baicalin inhibited calcium influx induced by TRPV4 activation. Cellular temperature elevated significantly when treated with GSK1016790A or hypaconitine, while the results were reversed when cells were treated with HC067047 or baicalin.Discussion and conclusions: The changes in cellular temperature are speculated to be caused by the alteration of intracellular calcium influx mediated by TRPV4. In addition, the 'cold/hot' properties of compounds in TCM can be classified by using cellular temperature detection.

Beta-elemene inhibits breast cancer metastasis through blocking pyruvate kinase M2 dimerization and nuclear translocation.

Pyruvate kinase M2 (PKM2), playing a central role in regulating aerobic glycolysis, was considered as a promising target for cancer therapy. However, its role in cancer metastasis is rarely known. Here, we found a tight relationship between PKM2 and breast cancer metastasis, demonstrated by the findings that beta-elemene (ß-elemene), an approved drug for complementary cancer therapy, exerted distinct anti-metastatic activity dependent on PKM2. The results indicated that ß-elemene inhibited breast cancer cell migration, invasion in vitro as well as metastases in vivo. ß-Elemene further inhibited the process of aerobic glycolysis and decreased the utilization of glucose and the production of pyruvate and lactate through suppressing pyruvate kinase activity by modulating the transformation of dimeric and tetrameric forms of PKM2. Further analysis revealed that ß-elemene suppressed aerobic glycolysis by blocking PKM2 nuclear translocation and the expression of EGFR, GLUT1 and LDHA by influencing the expression of importin α5. Furthermore, the effect of ß-elemene on migration, invasion, PKM2 transformation, and nuclear translocation could be reversed in part by fructose-1,6-bisphosphate (FBP) and L-cysteine. Taken together, tetrameric transformation and nuclear translocation of PKM2 are essential for cancer metastasis, and ß-elemene inhibited breast cancer metastasis via blocking aerobic glycolysis mediated by dimeric PKM2 transformation and nuclear translocation, being a promising anti-metastatic agent from natural compounds.

Acupuncture plus Chinese Herbal Medicine for Irritable Bowel Syndrome with Diarrhea: A Systematic Review and Meta-Analysis.

PURPOSE: To comprehensively evaluate the efficacy and safety of acupuncture combined with Chinese herbal medicine (CHM) in treating irritable bowel syndrome with diarrhea (IBS-D). METHODS: Relevant randomized controlled trials (RCTs) were systemically retrieved from electronic databases from inception to March 2018, including the Cochrane Central Register of Controlled Trials (CENTRAL), PubMed, EMBASE, China National Knowledge Infrastructure (CNKI), Chinese Biological Medical Database (CBM, SinoMed), China Science and Technology Journal Database (VIP), and Wan Fang Data. Meanwhile, pooled estimates, including the 95% confidence interval (CI), were calculated for primary and secondary outcomes of IBS-D patients. Besides, quality of relevant articles was evaluated using the Cochrane Collaboration's risk of bias tool, and the Review Manager 5.3 and Stata12.0 softwares were employed for analyses. RESULTS: A total of 21 RCTs related to IBS-D were included into this meta-analysis. Specifically, the pooled results indicated that (1) acupuncture combined with CHM might result in more favorable improvements compared with the control group (relative risk [RR] 1.29; 95% CI 1.24-1.35; P =0.03); (2) the combined method could markedly enhance the clinical efficacy in the meantime of remarkably reducing the scores of abdominal pain (standardized mean difference [SMD] -0.45; 95% CI -0.72, -0.17; P = 0.002), abdominal distention/discomfort (SMD -0.36; 95% CI -0.71, -0.01; P = 0.04), diarrhea (SMD -0.97; 95% CI -1.18, -0.75; P < 0.00001), diet condition (SMD -0.73; 95% CI -0.93, -0.52; P<0.00001), physical strength (SMD -1.25; 95% CI -2.32, -0.19; P = 0.02), and sleep quality (SMD -1.02; 95% CI -1.26, -0.77; P < 0.00001) compared with those in the matched groups treated with western medicine, or western medicine combined with CHM. Additionally, a metaregression analysis was constructed according to the name of prescription, acupuncture type, treatment course and publication year, and subgroup analyses stratified based on the names of prescriptions and acupoints location were also carried out, so as to explore the potential heterogeneities; and (3) IBS-D patients treated with the combined method only developed inconspicuous adverse events; more importantly, the combined treatment had displayed promising long-term efficacy. CONCLUSIONS: Findings in this study indicate that acupuncture combined with CHM is suggestive of an effective and safe treatment approach for IBS-D patients, which may serve as a promising method to treat IBS-D in practical application. However, more large-scale, multicenter, long-term, and high-quality RCTs are required in the future, given the small size, low quality, and high risk of the studies identified in this meta-analysis.

Protective Effects of Magnesium Glycyrrhizinate on Methotrexate-Induced Hepatotoxicity and Intestinal Toxicity May Be by Reducing COX-2.

Magnesium isoglycyrrhizinate (MgIG), which has been widely employed to treat chronic hepatitis, is synthesized from 18-ß glycyrrhizic acid, a main component of traditional Chinese medicine Glycyrrhiza uralensis Fisch. Although the protective effects of MgIG on methotrexate (MTX)-induced liver toxicity have been well-documented, the underlying mechanism remains elusive. MTX was initially used to treat pediatric acute leukemia, and has been widely applied to psoriasis therapy. However, its clinical applications are limited due to hepatotoxicity and intestinal toxicity. Herein, prophylactic administration of MgIG (9 and 18 mg/kg/day) significantly reduced the levels of aspartate aminotransferase and alanine aminotransferase in the serum of rats receiving intravenous injection of MTX (20 mg/kg body weight). MgIG also attenuated MTX-induced hepatic fibrosis. Moreover, it better protected against MTX-induced hepatocyte apoptosis and decreased the serum level of malondialdehyde than reduced glutathione (80 mg/kg/day) did. Interestingly, MTX-induced cyclooxygenase-2 (COX-2) expression, intestinal permeability and inflammation were attenuated after MgIG administration. In addition, MgIG (9 and 18 mg/kg) reduced MTX-induced colocalization of zonula occludens-1 (ZO-1) and connexin 43 (Cx43) in intestinal villi. In conclusion, MgIG exerted beneficial effects on MTX-induced hepatotoxicity and intestinal damage, as a potentially eligible drug for alleviating the hepatic and intestinal side effects of MTX during chemotherapy.

D-limonene exhibits anti-inflammatory and antioxidant properties in an ulcerative colitis rat model via regulation of iNOS, COX-2, PGE2 and ERK signaling pathways.

D-limonene has been demonstrated to have important immunomodulatory properties, including antitumor effects, and may alleviate asthma and allergies. In the present study, the anti­inflammatory effects of D­limonene were investigated in an ulcerative colitis (UC) rat model. Healthy male Sprague­Dawley rats were randomly divided into control, untreated UC, and treatment with 50 or 100 mg/kg D­limonene UC groups. In UC rats, disease activity and colonic mucosa damage were significantly reduced by the anti­inflammatory effects of D­limonene, via suppression of matrix metalloproteinase (MMP)­2 and ­9 gene expression. In addition, treatment with D­limonene significantly increased antioxidant, inducible nitric oxide synthase (iNOS) and cyclooxygenase­2 (COX­2) protein expression levels in UC rats. A decrease in prostaglandin E2 (PGE2) production, transforming growth factor­ß (TGF­ß) gene expression and an increase phosphorylated­extracellular signal regulated kinase (ERK) 1/2 expression levelswere observed in UC rats treated with D­limonene. In conclusion, D­limonene reduced MMP­2 and ­9 mRNA expression levels via regulation of the iNOS, COX­2, PGE2, TGF­ß and ERK1/2 signaling pathways in a UC rat model, indicating its potential antioxidant and anti­inflammatory properties.

Measuring the adherence energy of the resin-metal interface with two fracture mechanics methods: The DCB and NTP tests.

OBJECTIVE: To evaluate the abilities of the DCB and the NTP test for measuring adherence of an adhesive joint between a resin and a metal interface. METHODS: Two-hundred stainless steel metal beams (diam. 50×5×2 mm) were cast and treated by the following methods: (1) sandblasting with aluminum oxide, followed by treatment with (2) the Rocatec system or (3) the Alloy primer. Superbond and Panavia F 2.0 were used as adhesives. The fracture energy (G1C) and fracture toughness (K1C) of two adhesives were compared by two-way analysis of variance. RESULTS: With the DCB test, Superbond was more effective than Panavia, regardless of the surface treatment and conditions of crack propagation. The overall effectiveness of the treatments was in the following order: sandblasting + Rocatec > sandblasting alone > sandblasting + Alloy primer. The adherence energy in an aqueous medium was lower than that in air. With the NTP test, similar performances were obtained with three surface treatments. However, the potential of Rocatec seemed slightly higher. CONCLUSIONS: The DCB and NTP tests provide independent measures of the inherent value of an adhesive. Rocatec appeared to provide greater resistance of the bonded joints in an aqueous environment.

Preventive effect of Actinidia valvata Dunn extract on N-methyl-N'-nitro-N-nitrosoguanidine-induced gastrointestinal cancer in rats.

PURPOSE: This study was conducted to assess the preventive effect of Actinidia valvata Dunn (AVD) extract on an animal model of gastrointestinal carcinogenesis on the basis of changes in tumor incidence, cell proliferation, and apoptosis. MATERIALS AND METHODS: Seventy-five male Wistar rats were divided into five different treatment groups with 15 rats in each group. Group I was given normal feed, whereas Groups II to IV were treated with 10% sodium chloride in the first six weeks and 100 ug/mL of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in drinking water for 24 weeks. Group II was then given normal feed, whereas Group III was given AVD extract (0.24 g/kg/day) for 12 weeks. Group IV was given AVD extract from the first week to the 36th week, whereas Group V was treated with AVD extract alone for 36 weeks. All rats were sacrificed at the end of the 36-week experiment and assessed for the presence of gastrointestinal tumors. The occurrence of cancer was evaluated by histology. Bax, Bcl-2, Caspase-3, and cyclinD1 were determined by immunohistochemical staining and Western blotting. RESULTS: The incidences of gastric cancer were 0% in Group I, 73.3% in Group II, 33.3% in Group III, 26.7% in Group IV, and 0% in Group V. Bcl-2 and cyclinD1 expression was decreased in AVD extract treated groups, whereas Bax and Caspase-3 expression was increased. Comparison with group II revealed significant differences (p<0.01). CONCLUSIONS: AVD extract exhibits an obvious preventive effect on gastrointestinal carcinogenesis induced by MNNG in rats through the regulation of cell proliferation and apoptosis.

[Effect of Fagopyrum cymosum (Trev.) Meisn alcohol extract on defecation and isolated colon of diarrhea-IBS rats and its mechanism].

OBJECTIVE: To explore the intervention of Fagopyrum cymosum (Trev.) Meisn alcohol extract (FAE) on defecation function and motor functions of isolated colons of diarrhea-predominant irritable bowel syndrome (D-IBS) rats and to study its underlying mechanism. METHODS: The D-IBS rat model was established by neonatal pups maternal separation (NMS) combined with intracolonic infusion of acetic acid (AA). Adult IBS rats were randomly divided into the pre-intervention control group (n = 10, with no gastrogavage), the normal saline control group (n = 10, administered with normal saline by gastrogavage), the pre-treatment model group (n = 8,with no gastrogavage),the normal saline model group (n = 8, administered with normal saline by gastrogavage), the low dose FAE group (n = 8, administered with 6 g/kg FAE by gastrogavage), the high dose FAE group (n = 8, administered with 24 g/kg FAE by gastrogavage), and the Pinaverium Bromide group (n = 8, administered with 0.02 g/kg Pinaverium Bromide by gastrogavage). All medication was performed once daily for 2 weeks. The abdominal withdrawal reflex (AWR) was employed to evaluate the visceral hypersensitivity; their loose and watery stool grade was assessed by Bristol scores for stool consistency; and their fresh feces weight was calculated. In vitro effect of different concentrations of FAE and Pinaverium Bromide (0.02 µg/mL) on spontaneous contraction and spasmodic contraction induced by acetylcholine (Ach) in rats' isolated colon were observed and the influence on the intestinal calcium channel was evaluated. RESULTS: Compared with the pre-intervention control group, the pain pressure threshold and the maximum tolerance pressure decreased significantly in the pre-intervention model group (P < 0.05), and the loose and watery stool grade and fresh feces weight increased drastically (P < 0.01). Compared with the normal saline control group, the pain pressure threshold and the maximum tolerance pressure decreased significantly in the normal saline model group (P < 0.05), and the loose and watery stool grade and fresh feces weight increased markedly (P < 0.01). Compared with the normal saline model group, the pain pressure threshold of 24 g/kg FAE and Pinaverium Bromide group significantly increased (P < 0.05). The loose and watery stool grade and fresh feces weight decreased obviously in the low dose FAE group, the high dose FAE group, and the Pinaverium Bromide group (P < 0.05). FAE (30, 100, 300, 1,000, and 3,000 µg/mL) and Pinaverium Bromide could significantly inhibit spontaneous contraction of isolated intestines (P < 0.05, P < 0.01), and FAE (30, 100, and 300 x 10(-6) g/mL) could remarkably inhibit their spasmodic contraction and contractile tension induced by Ach and Ca2+ respectively (P < 0.05, P < 0.01) in a concentration-dependent manner. Pinaverium Bromide also could significantly inhibit Ach and Ca2+ induced contraction. CONCLUSION: Effective components of FAE improved the defecation function and inhibited enterospasm induced intestinal hyperactivity in IBS model rats via antagonizing calcium channel competitively and inhibiting colonic motility dose-dependently.

In Vivo and In Vitro Antinociceptive Effect of Fagopyrum cymosum (Trev.) Meisn Extracts: A Possible Action by Recovering Intestinal Barrier Dysfunction.

Fagopyrum cymosum (Trev.) Meisn (Fag) is a herb rhizome which has been widely used to treat diseases. To investigate the effects and mechanisms of the Fag on irritable bowel syndrome (IBS), in vivo neonatal pups maternal separation (NMS) combined with intracolonic infusion of acetic acid (AA) was employed to establish IBS rat models. Fag reduced their visceral hyperalgesia and the whole gut permeability, ameliorated colonic mucosa inflammation and injury, and upregulated the expression of decreased tight junction proteins (TJs) of claudin-1, occludin, and ZO-1 (except ZO-2) in colonic epithelium. Caco-2 monolayer cells were incubated with TNF-α and IFN-γ in vitro to establish an epithelial barrier dysfunction model whose transepithelial electrical resistance (TER) depended more on dose of Fag than that of the controls, and whose TJs levels were lower than those of the controls. Fag upregulated the NP-40 insoluble and soluble components of the four TJs markedly in a dose-dependent manner. These data suggest that Fag alleviated the hyperalgesia of IBS rats by reducing intestinal inflammation and enhancing mucosal epithelial function after regulating the structure and function of TJs.

Cryptotanshinone has diverse effects on cell cycle events in melanoma cell lines with different metastatic capacity.

PURPOSE: Cryptotanshinone is a major active component of Salvia miltiorrhiza, which is often used as Chinese herbal medicine in cancer therapy. Here, we systematically assessed the anti-tumor effect of Cryptotanshinone on two melanoma cell lines with low/high-metastatic capacity (B16/B16BL6). METHODS: MTT and LDH assays were used to evaluate cell growth and cytotoxicity. We assessed the effect of Cryptotanshinone on cell apoptosis or proliferation by Annexin V, TUNEL or BrdU assay. Cell cycle distribution was detected by flow cytometry. The integrity of cell cycle checkpoints was determined by mutational analyses of B-RAF and N-RAS, and the expression of cell cycle-associated proteins by western blotting. RESULTS: Treatment with Cryptotanshinone had no obvious effect on cell apoptosis but significantly inhibited cell proliferation. Cryptotanshinone slightly increased the expression of p53, Chk1, and Chk2 in both B16 and B16BL6. Interestingly, Cryptotanshinone induced G1 arrest with a concomitant increase in p21 expression in B16BL6 cells. However, in B16 cells, Cryptotanshinone induced the G2/M arrest through its induction of Cdc25c. Regulation of Cyclin A1, Cyclin B1 and Cdk1/cdc2 expression might contribute to the different cell cycle patterns in B16 and B16BL6 after Cryptotanshinone treatment. CONCLUSIONS: Cryptotanshinone could have diverse effects on cell cycle events in melanoma cell lines with different metastatic capacity. This property might offer an opportunity to study underlying mechanisms for the different antitumor effects of administered Cryptotanshinone in B16 and B16BL6 cells.