Effects of Chinese Fructus Mume Formula and Its Separated Prescription Extract on Insulin Resistance in Type 2 Diabetic Rats

The effect of Fructus Mume formula and its separated prescription extract on insulin resis- tance in type 2 diabetic rats was investigated. The rat model of type 2 diabetes was established by feed- ing on a high-fat diet for 8 weeks and by subsequently intravenous injection of small doses of strepto- zotocin. Rats in treatment groups, including the Fructus Mume formula treatment group （FM）, the cold property herbs of Fructus Mume formula treatment group （CFM）, the warm property herbs of Fructus Mume formula treatment group （WFM）, were administrated with Fructus Mume formula and its sepa- rated prescription extract by gavage, while the rats in diabetic model group （DM） and metformin group （MET） were given by gavage with normal saline and metformin correspondingly. The body weight be- fore and after treatment was measured, and the oral glucose tolerance test （OGTT） and the insulin re- lease test （IRT） were performed. The homeostasis model assessment-insulin resistance index （HOMA-IR） was calculated. The protein and mRNA expression levels of Insr, β-arrestin-2, Its-1 and Glut-4 in the liver, skeletal muscle and fat tissues were detected by using Western blotting and RT-PCR respectively. The results demonstrated that, as compared with DM group, OGTT, IRT （0 h, 1 h） levels and HOMR-IR in treatment groups were all reduced, meanwhile their protein and mRNA expression levels of Insr, Irs-1 and Glut-4 in the liver, skeletal muscle and fat tissues were obviously increased, and their protein and mRNA expression levels of β-arrestin-2 in the liver and skeletal muscle tissues were also markedly increased. It was suggested that the Fructus Mume formula and its separated prescription extracts could effectively improve insulin resistance in type 2 diabetic rats, which might be related to the up-regulated expression of Insr, Irs-1 and Glut-4 in the liver, skeletal muscle and fat tissues, and β-arrestin-2 in the liver and skeletal muscle tissues.

Study on mechanism of Fructus mume in treatment of colorectal cancer based on network pharmacology method

Objective:To explore the key targets and mechanisms of action of Mume Fructus in the treatment of colorectal cancer based on network pharmacology.Methods:TCMSP database was adopted to screen the active components and potential targets of Mume Fructus.GeneCards and Disgest databases were used to collect disease targets for colorectal cancer.Cytoscape software was used to construct the component-target network diagram.STRING database was used to draw the PPI network.Finally,DAVID database was used to perform GO biological process analysis and KEGG pathway enrichment analysis of centre target proteins.Results:There were 58 potential targets for the treatment of colorectal cancer in Mume Fructus.The protein interaction network suggested that TP53、AKT1、MAPK8、JUN、EGF and TNF may be the core targets for the treatment of colorectal cancer in Mume Fructus.Gene ontology enrichment analysis showed 108 cellular biological processes,and KEGG pathway enrichment analysis showed 88 related signaling pathways,including p53,TNF,HIF-1 and so on.Conclusion:The treatment of colorectal cancer by Mume Fructus may be multi-target,multi-channel,and multi-level.This consequence may provide ideas and a basis for further research.

Protective effect of Fructus Mume total flavone against SH-SY5Y cells damage induced by MPP+and its mechanism

Objective:To investigate the neuroprotective effects of Fructus Mume total flavone(FMF)against cell apoptosis and mitochondrial injury induced by 1-methyl-4-phenylpyridinium(MPP^(+))in human neuroblastoma(SH-SY5Y)cells and explore its molecular mechanisms.Methods:MPP^(+) induced SH-SY5Y cells injury model were established in vitro cell culture,the cells were divided into 5 groups:normal control group,model group(250μmol·L^(-1) MPP^(+)),FMF low-and middle-and high-dose experimental group(10,50,100μmol·L^(-1) FMF).After 72 h administration,4’,6-diamidino-2-phenylindole(DAPI)staining was used to observe the effects of different concentrations of FMF on the morphologic changes of apoptotic cells,the ratio of cell apoptosis was measured by Annexin-FITC/PI double staining.The mitochondrial membrane electro-bit were detected by flow cytometry(FCM).The expression of Bcl-2,Bax and Caspase-3 were detected by Western blot.Results:The results of DAPI staining showed that the injury SH-SY5Y cells induced by MPP+were densely condensed,the nucleus showed nuclear shrinkage,showing an apoptotic characteristic morphology;after 72h of FMF action,the apoptotic morphology of the cells showed different degrees of improvement,and the apoptotic number of SH-SY5Y cells also decreased.Compared with that in the normal control group,the apoptotic rate and of mitochondrial membrane electrobit of SH-SY5Y cells in the model group increased significantly(P<0.01),the expression of Bax and Caspase-3 proteins increased significantly(P<0.01),Bcl-2 protein and the ratio of Bcl-2/Bax decreased significantly(P<0.01).Compared with that in the model group,the apoptotic rate and mitochondrial membrane electro-bit of SH-SY5Y cells in FMF groups(10,50,100μmol·L^(-1))were significantly lower,while Bax and Caspase-3 proteins were significantly lower(P<0.01),and Bcl-2 protein and the ratio of Bcl-2/Bax were significantly higher,with statistically significant difference in FMF middle-and high-dose experimental groups(P<0.01).The results indicated that FMF can decrease the experession level of Bax and Caspase-3 and increase the ratio of Bcl-2/Bax,inhibit MPP+induced apoptosis.Conclusion:FMF improves the damage of SH-SY5Y cells induced by MPP+,and plays a neuroprotective effect by regulating the expressions of related proteins in mitochondrial apoptosis pathway.

Huanglian decoction suppresses the growth of hepatocellular carcinoma cells by reducing CCNB1 expression

BACKGROUND Hepatocellular carcinoma(HCC)is one of the most prevalent cancers in human populations worldwide.Huanglian decoction is one of the most important Chinese medicine formulas,with the potential to treat cancer.AIM To investigate the role and mechanism of Huanglian decoction on HCC cells.METHODS To identify differentially expressed genes(DEGs),we downloaded gene expression profile data from The Cancer Genome Atlas Liver Hepatocellular Carcinoma and Gene Expression Omnibus(GSE45436)databases.We obtained phytochemicals of the four herbs of Huanglian decoction from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.We also established a regulatory network of DEGs and drug target genes and subsequently analyzed key genes using bioinformatics approaches.Furthermore,we conducted in vitro experiments to explore the effect of Huanglian decoction and to verify the predictions.In particular,the CCNB1 gene was knocked down to verify the primary target of this decoction.Through the identification of the expression levels of key proteins,we determined the primary mechanism of Huanglian decoction in HCC.RESULTS Based on the results of the network pharmacological analysis,we revealed 5 bioactive compounds in Huanglian decoction that act on HCC.In addition,a protein-protein interaction network analysis of the target genes of these five compounds as well as expression and prognosis analyses were performed in tumors.CCNB1 was confirmed to be the primary gene that may be highly expressed in tumors and was significantly associated with a worse prognosis.We also noted that CCNB1 may serve as an independent prognostic indicator in HCC.Moreover,in vitro experiments demonstrated that Huanglian decoction significantly inhibited the growth,migration,and invasiveness of HCC cells and induced cell apoptosis and G2/M phase arrest.Further analysis showed that the decoction may inhibit the growth of HCC cells by downregulating the CCNB1 expression level.After Huanglian decoction treatment,the expression levels of Bax,caspase 3,caspase 9,p21 and p53 in HCC cells were increased,while the expression of CDK1 and CCNB1 was significantly decreased.The p53 signaling pathway was also found to play an important role in this process.CONCLUSION Huanglian decoction has a significant inhibitory effect on HCC cells.CCNB1 is a potential therapeutic target in HCC.Further analysis showed that Huanglian decoction can inhibit HCC cell growth by downregulating the expression of CCNB1 to activate the p53 signaling pathway.