Enterolactone and trabectedin suppress epithelial ovarian cancer synergistically via upregulating THBS1.

Epithelial ovarian cancer (EOC) is the most common and fatal subtype of ovarian malignancies, with no effective therapeutics available. Our previous studies have demonstrated extraordinary suppressive efficacy of enterolactone (ENL) on EOC. A chemotherapeutic agent, trabectedin (Trabe), is shown to be effective on ovarian cancer, especially when combined with other therapeutics, such as pegylated liposomal doxorubicin or oxaliplatin. Thrombospondin 1 (THBS1), a kind of matrix glycoprotein, plays important roles against cancer development through inhibiting angiogenesis but whether it is involved in the suppression of EOC by ENL or Trabe remains unknown. To test combined suppressive effects of ENL and Trabe on EOC and possible involvement of THBS1 in the anticancer activities of ENL and Trabe. The EOC cell line ES-2 was transfected with overexpressed THBS1 by lentivirus vector. We employed tube formation assay to evaluate the anti-angiogenesis activity of ENL and of its combined use with Trabe after THBS1 overexpression and established drug intervention and xenograft nude mouse cancer models to assess the in vivo effects of the hypothesized synergistic suppression between the agents and the involvement of THBS1. Mouse fecal samples were collected for 16S rDNA sequencing and microbiota analysis. We detected strong inhibitory activities of ENL and Trabe against the proliferation and migration of cancer cells and observed synergistic effects between ENL and Trabe in suppressing EOC. ENL and Trabe, given either separately or in combination, could suppress the tube formation capability of human microvascular endothelial cells, and this inhibitory effect became even stronger with THBS1 overexpression. In the ENL plus Trabe combination group, the expression of tissue inhibitor of metalloproteinases 3 and cluster of differentiation 36 was both upregulated, whereas matrix metalloproteinase 9, vascular endothelial growth factor, and cluster of differentiation 47 were all decreased. With the overexpression of THBS1, the results became even more pronounced. In animal experiments, combined use of ENL and Trabe showed superior inhibitory effects to either single agent and significantly suppressed tumor growth, and the overexpression of THBS1 further enhanced the anti-cancer activities of the drug combination group. ENL and Trabe synergistically suppress EOC and THBS1 could remarkably facilitate the synergistic anticancer effects of ENL and Trabe.

Fecal microbiota transplantation inhibits colorectal cancer progression: Reversing intestinal microbial dysbiosis to enhance anti-cancer immune responses.

Many lines of evidence demonstrate the associations of colorectal cancer (CRC) with intestinal microbial dysbiosis. Recent reports have suggested that maintaining the homeostasis of microbiota and host might be beneficial to CRC patients, but the underlying mechanisms remain unclear. In this study, we established a CRC mouse model of microbial dysbiosis and evaluated the effects of fecal microbiota transplantation (FMT) on CRC progression. Azomethane and dextran sodium sulfate were used to induce CRC and microbial dysbiosis in mice. Intestinal microbes from healthy mice were transferred to CRC mice by enema. The vastly disordered gut microbiota of CRC mice was largely reversed by FMT. Intestinal microbiota from normal mice effectively suppressed cancer progression as assessed by measuring the diameter and number of cancerous foci and significantly prolonged survival of the CRC mice. In the intestine of mice that had received FMT, there were massive infiltration of immune cells, including CD8+ T and CD49b+ NK, which is able to directly kill cancer cells. Moreover, the accumulation of immunosuppressive cells, Foxp3+ Treg cells, seen in the CRC mice was much reduced after FMT. Additionally, FMT regulated the expressions of inflammatory cytokines in CRC mice, including down-regulation of IL1a, IL6, IL12a, IL12b, IL17a, and elevation of IL10. These cytokines were positively correlated with Azospirillum_sp._47_25, Clostridium_sensu_stricto_1, the E. coli complex, Akkermansia, Turicibacter, and negatively correlated with Muribaculum, Anaeroplasma, Candidatus_Arthromitus, and Candidatus Saccharimonas. Furthermore, the repressed expressions of TGFb, STAT3 and elevated expressions of TNFa, IFNg, CXCR4 together promoted the anti-cancer efficacy. Their expressions were positively correlated with Odoribacter, Lachnospiraceae-UCG-006, Desulfovibrio, and negatively correlated with Alloprevotella, Ruminococcaceae UCG-014, Ruminiclostridium, Prevotellaceae UCG-001 and Oscillibacter. Our studies indicate that FMT inhibits the development of CRC by reversing gut microbial disorder, ameliorating excessive intestinal inflammation and cooperating with anti-cancer immune responses.

Systematic review and meta-analysis of Coptis chinensis Franch.-containing traditional Chinese medicine as an adjunct therapy to metformin in the treatment of type 2 diabetes mellitus.

Background: In China, Coptis chinensis Franch. (Chinese name: Huanglian) prescriptions (HLPs) are prominent hypoglycemic agents used in glycemic control. However, the curative effect of HLPs as adjunctive therapies for type 2 diabetes mellitus (T2DM) has not been evaluated. Based on a systematic review and a meta-analysis, this study was conducted to assess the effects of HLPs combined with metformin as a reinforcing agent for T2DM. Materials and methods: A total of 33 randomized controlled trials (RCTs) reporting on 2,846 cases concerning the use of HLPs in the treatment of T2DM were identified from the China National Knowledge Infrastructure (CNKI), Weipu (VIP), Wanfang, PubMed, Cochrane Library, and EMBASE databases. Primary outcomes included fasting blood glucose (FBG), 2-h postprandial blood glucose (2hPG), glycosylated hemoglobin, type A1c (HbA1c), fasting serum insulin (FINS), and homeostasis model assessment of insulin resistance (HOMA-IR). Secondary outcomes included total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL-c), and gastrointestinal dysfunction (GD). Continuous data were expressed as mean differences (MDs) with 95% confidence intervals (CIs). The methodological quality of the included RCTs was assessed by Cochrane evidence-based medicine systematic evaluation. Statistical analysis was performed using the Review Manager and Stata software. The required information size and treatment benefits were evaluated by trial sequential analysis (TSA). The quality of evidence was rated using the Grades of Recommendation Assessment, Development, and Evaluation (GRADE) approach. Results: The results revealed that HLPs are beneficial to improve the following: FBG (MD = -1.16%, 95% CI: -1.24 to -1.07), 2hPG (MD = -1.64%, 95% CI: -1.84 to -1.43), HbA1c (MD = -0.78%, 95% CI:-0.96 to -0.60), FINS (MD = -1.94%, 95% CI: -2.68 to -1.20), HOMA-IR (MD = -0.77%, 95% CI: -1.28 to -0.27), TC (MD = -0.70%, 95% CI: -1.00 to -0.39), TG (MD = -0.57%, 95% CI: -0.74 to -0.40), LDL-c (MD = -0.70%, 95% CI: -0.97 to -0.43), and HDL-c (MD = -0.21%, 95% CI: -0.32 to -0.10) for patients with T2DM. The funnel plot, Egger's test, and trim-and-fill method indicated a moderate publication bias in the results. The TSA showed that the required sample size of HLPs in improving FBG, 2hPG, HbA1c, FINS, HOMA-IR, TC, TG, LDL-c, and HDL-c could sufficiently draw reliable conclusions. GRADE assessment revealed that the quality of the evidence for the effectiveness of HLPs in improving FBG was moderate, but the quality of evidence for 2hPG, HbA1c, FINS, HOMA-IR, TC, TG, LDL-c, and HDL-c was low, and for GD was very low. Conclusion: The systematic review and meta-analysis suggested that HLPs were beneficial for achieving glycemic control. However, HLPs recommended for T2DM patients have yet to be confirmed because of the poor methodological quality of some trials. Therefore, more RCTs with multicenter and double-blind designs are needed to assess the efficacy of HLPs for patients with T2DM.

Corrigendum: Systematic review and meta-analysis of Coptis Chinensis Franch. -containing traditional Chinese medicine as an adjunct therapy to metformin in the treatment of type 2 diabetes mellitus.

[This corrects the article DOI: 10.3389/fphar.2022.956313.].

TMT-based proteomics analysis reveals the efficacy of jiangzhuo formula in improving the lipid profiles of dyslipidemia rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Jiangzhuo Formula (JZF) is a traditional Chinese herbal prescription that is clinically applied to treat dyslipidemia. However, the mechanism underlying its efficacy remains unexplored. AIM OF THE STUDY: This study aims to elucidate the underlying mechanisms, explore potential pathways, and identify the key proteins of JZF for the treatment of dyslipidemia. METHODS: In this work, Q-Orbitrap high-resolution liquid chromatography mass spectrometry was used to identify the natural ingredients in JZF, rats with dyslipidemia were established via a high-fat diet for four weeks, then the dyslipidemia rats were treated with high-dose JZF (9 g/d) and low-dose JZF (4.5 g/d) for four weeks. After treatment, serum lipid detection and Oil-red-O staining were conducted to assess the efficacy of JZF in ameliorating dyslipidemia. Tandem mass tag (TMT) -based quantitative proteomics technology was then used to evaluate the roles and importance of proteins from the extracted hepatic tissue. The differentially expressed proteins were assessed by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, Gene Ontology (GO), and protein-protein interaction (PPI) networks. Western blot and PCR analysis were used to validate the potential targets regulated by JZF. RESULTS: JZF could significantly improve the blood lipid profiles of serum and fat deposits of the liver. A total of 123 differentially expressed proteins were detected after JZF intervention, comprising 65 up-regulated proteins and 58 down-regulated proteins. The KEGG pathway analysis revealed that cholesterol metabolism, the PPAR signaling pathway, and bile secretion were the principal pathways involved in the disordered lipid metabolism, while GO analysis suggested that proteins that are located in the cell, regulate cellular processes, and show binding activity contribute to reductions in lipids. The combination of proteomics, Western blot, and PCR suggested that Apolipoprotein B (APOB), Apolipoprotein E (APOE), cholesterol 7 alpha-hydroxylase A1 (CYP7A1), and Hydroxymethylglutaryl-CoA synthase 1 (HMGCS1) might play critical roles in JZF's lipid-lowering network. CONCLUSION: JZF can effectively improve lipid profiles via multiple pathways involved in cholesterol metabolism, the PPAR signaling pathway, and bile secretion. Generally, the proteomics techniques used in this research show that JZF could be a promising drug for the treatment of dyslipidemia.

Network pharmacology and metabolomics study on the intervention of traditional Chinese medicine Huanglian Decoction in rats with type 2 diabetes mellitus.

ETHNOPHARMACOLOGICAL RELEVANCE: Type 2 diabetes mellitus (T2DM) is currently one of the most prominent and global chronic conditions. Huanglian Decoction (HLD) is a traditional Chinese medicine (TCM) preparation that has been used to treat T2DM for thousands of years in China. However, its mechanism of action at the metabolic level is still unclear. The purpose of this work is to study the mechanism of HLD in treating T2DM based on metabolomics and network pharmacology. MATERIALS AND METHODS: In this study, metabolomics combined with network pharmacology was used to elucidate the therapeutic mechanism of HLD in T2DM. Serum samples were collected from rats with T2DM, induced by a high-sugar and high-fat diet combined with streptozotocin (STZ), to measure the levels of biochemical markers. Urinary metabolomics-based analysis using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) was conducted to evaluate the differential metabolites from multiple metabolic pathways. RESULTS: After treatment with HLD for 4 weeks, biochemical indicators, including fasting blood glucose (FBG), blood lipid, fasting insulin (FINS), insulin sensitivity index (ISI), and homeostasis model assessment of insulin resistance (HOMA-IR), were significantly improved. Metabolomics results revealed that HLD regulated the biomarkers, such as cytosine, L-carnitine, betaine, phenylalanine, glucose, citrate, phenylpyruvate, and hippuric acid in glyoxylate and dicarboxylate metabolism, phenylalanine metabolism, and tricarboxylic acid (TCA) cycle. The combination of network pharmacology, metabolomics, western blot, and PCR showed that HLD can treat T2DM by enhancing the gene and protein expression levels of glucose transporter 4 (GLUT4), insulin receptor (INSR), and mitogen-activated protein kinase 1 (MAPK1) to interfere with glyoxylate and dicarboxylate metabolism. CONCLUSIONS: The study based on metabolomics and network pharmacology indicated that HLD can improve T2DM through multiple targets and pathways, and it may be a useful alternative therapy for the treatment of T2DM.

Urinary Metabolomics Study of the Intervention Effect of Hypoglycemic Decoction on Type 2 Diabetes Mellitus Rats Model.

The hypoglycemic decoction (HD) is a traditional Chinese medicine (TCM) preparation for the treatment of diabetes mellitus (DM), with a remarkable therapeutic effect. However, its mechanism of action is still unclear at the metabolic level. In this study, the biochemical markers from type 2 DM (T2DM) rats, induced by a high-sugar and high-fat diet combined with streptozotocin (STZ), were detected. The metabolomics-based analysis using high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) was conducted to evaluate urine samples from control, model, metformin, and HD groups. After oral administration of HD for 28 days, the general state, weight, fasting blood glucose (FBG), blood lipid level, oral glucose tolerance test (OGTT), fasting insulin (FINS), insulin sensitivity index (ISI), and homeostasis model assessment of insulin resistance (HOMA-IR) were significantly improved (P < 0.01). The western blotting showed that HD can enhance the protein expression of glucose transporter 4 (GLUT4) and adenosine monophosphate-activated protein kinase (AMPK). The metabolomics results revealed that after treatment with HD, the levels of L-carnitine, 1-methyladenosine, 1-methylhistamine, and 3-indoleacrylic acid were upregulated and the levels of riboflavin, phenylalanine, atrolactic acid, 2-oxoglutarate, citrate, isocitrate, cortisol, and glucose were downregulated. The main mechanism may be closely related to the regulation of the tricarboxylic acid (TCA) cycle, phenylalanine metabolism, glyoxylate metabolism, and dicarboxylate metabolism. Additionally, it was also found that HD can regulate the protein expression of GLUT4 and AMPK to interfere with TCA cycle and carbohydrate metabolism to treat T2DM.

Ginsenoside Rg3 (Shenyi Capsule) Combined with Chemotherapy for Digestive System Cancer in China: A Meta-Analysis and Systematic Review.

OBJECTIVE: In China, ginsenoside Rg3 is often used in combination with chemotherapy to treat digestive system cancer. We here performed a meta-analysis and systematic review to provide a much needed high-quality evaluation of the efficacy and safety of ginsenoside Rg3 combined with chemotherapy in these cancers. MATERIALS AND METHODS: The PubMed, EMBASE, Cochrane Library, China National Knowledge Infrastructure (CNKI), Wanfang, and Weipu (VIP) databases were searched. All randomized controlled trials (RCTs) concerning ginsenoside Rg3 combined with chemotherapy for digestive system cancer were selected. Dichotomous data were expressed as odds ratios (ORs) with 95% confidence intervals (CI). The methodological quality of the included studies was evaluated according to the Cochrane evidence-based medicine system, and the statistical analyses were performed with Review Manager 5.3 and STATA 12.0 software. In addition, the Grades of Recommendation Assessment, Development and Evaluation (GRADE) approach was used to rate the quality of the evidence. Trial sequential analysis (TSA) was used to evaluate information size and treatment benefits. RESULTS: A total of 18 trials comprising 1531 patients were included in this study. The results revealed that the trials were of sufficient standard to draw reliable conclusions that ginsenoside Rg3 combined with chemotherapy could improve the objective response rate (ORR; OR 2.17, 95% CI 1.72-2.73), disease control rate (DCR; OR 2.83, 95% CI 2.02-3.96), 1-year survival rate (SR; OR = 2.33, 95% CI = 1.24-4.37), Karnofsky Performance Scale (KPS; OR 2.67, 95% CI 1.76-4.03), gastrointestinal dysfunction (OR 0.44, 95% CI 0.31-0.61), and the decline of leucocyte count (OR 0.28, 95% CI 0.21-0.38). CONCLUSION: Ginsenoside Rg3 combined with chemotherapy can improve the clinical efficacy and alleviate treatment-induced side effects for digestive system cancer.

[Regulatory effect of lipo tiaozhi capsule on the expression of peroxisome proliferator-activated receptors mRNA in dyslipidemia rats and ApoE(-/-) mice].

OBJECTIVE: To study the regulatory effect of lipi tiaozhi capsule (LTC) on the expression of peroxisome proliferator-activated receptor (PPAR) alpha and gamma mRNA in dyslipidemia rats and ApoE(-/-) mice, and to explore its mechanisms for regulating lipid metabolism. Methods 48 Wistar rats were randomly divided into the blank group, the model group, the treatment group (treated by LTC), and the control group (treated by Xuezhi-kang Capsule). After four-week modeling (except the blank group) 30 ApoE(-/-) mice were randomly divided into the blank group, the treatment group, and the control group. LTC was given by gastrogavage to rats and ApoE(-/-) mice in LTC groups while XZKC was given to XZKC groups. The medication was conducted once daily for eight weeks. The serum TC and TG contents of rats and mice were determined by enzymic method. The serum high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were detected by precipitation method. PPARalpha and gamma mRNA expressions were detected in the liver tissue of the rats and mice by fluorescent PCR. RESULTS: Compared with the model group and the blank group, the serum contents of TC, TG, and LDL-C of rats or mice in the treatment group decreased significantly (P < 0.01). The serum content of HDL-C increased significantly (P < 0.01). PPARalpha and gamma mRNA expressions of rats or mice increased significantly (P < 0.01). Compared with the control group, the serum contents of TC, TG, and LDL-C of mice and rats in the treatment group decreased (all P < 0.05), the serum content of HDL-C increased significantly (P < 0.05, P < 0.01). And PPARalpha and gamma mRNA expressions of rats or mice increased significantly (P < 0.05). CONCLUSION: LTC could significantly increase PPARa and y mRNA expressions of experimental dyslipidemia rats and ApoE(-/-) mice, playing roles in regulating nuclear factors and further effecting lipid metabolism.