Zuogui Jiangtang Qinggan Formula improves glucolipid metabolism in type 2 diabetes mellitus complicated with non-alcoholic fatty liver disease by regulating FoxO1/MTP/APOB signaling pathway / 中国中药杂志

This study aimed to investigate the effect and mechanism of Zuogui Jiangtang Qinggan Formula(ZGJTQG) on the glucolipid metabolism of type 2 diabetes mellitus(T2DM) complicated with non-alcoholic fatty liver disease(NAFLD). NAFLD was induced by a high-fat diet(HFD) in MKR mice(T2DM mice), and a model of T2DM combined with NAFLD was established. Forty mice were randomly divided into a model group, a metformin group(0.067 g·kg~(-1)), and high-and low-dose ZGJTQG groups(29.64 and 14.82 g·kg~(-1)), with 10 mice in each group. Ten FVB mice of the same age were assigned to the normal group. Serum and liver tissue specimens were collected from mice except for those in the normal and model groups after four weeks of drug administration by gavage, and fasting blood glucose(FBG) and fasting insulin(FINS) levels were measured. The levels of total cholesterol(TC), triglyceride(TG), and low-density lipoprotein(LDL) were detected by the single reagent GPO-PAP method. Very low-density lipoprotein(VLDL) was detected by enzyme-linked immunosorbent assay(ELISA). Alanine aminotransferase(ALT) and aspartate ami-notransferase(AST) were determined by the Reitman-Frankel assay. The pathological changes in the liver were observed by hematoxylin-eosin(HE) staining and oil red O staining. Real-time fluorescence-based quantitative polymerase chain reaction(real-time PCR) and Western blot were adopted to detect the mRNA and protein expression of forkhead transcription factor O1(FoxO1), microsomal triglyceride transfer protein(MTP), and apolipoprotein B(APOB) in the liver. The results showed that high-dose ZGJTQG could signi-ficantly reduce the FBG and FINS levels(P<0.05, P<0.01), improve glucose tolerance and insulin resistance(P<0.05, P<0.01), alleviate the liver damage caused by HFD which was reflected in improving liver steatosis, and reduce the serum levels of TC, TG, LDL, VLDL, ALT, and AST(P<0.05, P<0.01) in T2DM mice combined with NAFLD. The findings also revealed that the mRNA and protein expression of FoxO1, MTP, and APOB in the liver was significantly down-regulated after the intervention of high-dose ZGJTQG(P<0.05, P<0.01). The above study showed that ZGJTQG could effectively improve glucolipid metabolism in T2DM combined with NAFLD, and the mechanism was closely related to the regulation of the FoxO1/MTP/APOB signaling pathway.

Zuogui Jiangtang Qinggan Prescription promotes recovery of intestinal mucosal barrier in mice with type 2 diabetes mellitus and nonalcoholic fatty liver disease by improving intestinal flora homeostasis / 中国中药杂志

This study aimed to investigate the recovery effect of Zuogui Jiangtang Qinggan Prescription on intestinal flora homeostasis control and intestinal mucosal barrier in type 2 diabetes mellitus(T2DM) with nonalcoholic fatty liver disease(NAFLD) induced by a high-fat diet. NAFLD was established in MKR transgenic mice(T2DM mice) by a high-fat diet(HFD), and subsequently treated for 8 weeks with Zuogui Jiangtang Qinggan Prescription(7.5, 15 g·kg~(-1)) and metformin(0.067 g·kg~(-1)). Triglyceride and liver function were assessed using serum. The hematoxylin-eosin(HE) staining and Masson staining were used to stain the liver tissue, while HE staining and AB-PAS staining were used to stain the intestine tissue. 16S rRNA sequencing was utilized to track the changes in the intestinal flora of the mice in each group. Polymerase chain reaction(PCR) and immunofluorescence were used to determine the protein and mRNA expression levels of ZO-1, Occludin, and Claudin-1. The results demonstrated that Zuogui Jiangtang Qinggan Prescription increased the body mass of T2DM mice with NAFLD and decreased the hepatic index. It down-regulated the serum biomarkers of liver function and dyslipidemia such as alanine aminotransferase(ALT), aspartate transaminase(AST), and triglycerides(TG), increased insulin sensitivity, and improved glucose tolerance. According to the results of 16S rRNA sequencing, the Zuogui Jiangtang Qinggan Prescription altered the composition and abundance of the intestinal flora, increasing the relative abundances of Muribaculaceae, Lactobacillaceae, Lactobacillus, Akkermansia, and Bacteroidota and decreasing the relative abundances of Lachnospiraceae, Firmicutes, Deslfobacteria, Proteobacteria, and Desulfovibrionaceae. According to the pathological examination of the intestinal mucosa, Zuogui Jiangtang Qinggan Prescritpion increased the expression levels of the tight junction proteins ZO-1, Occludin, and Claudin-1, promoted intestinal mucosa repair, protected intestinal villi, and increased the height of intestinal mucosa villi and the number of goblet cells. By enhancing intestinal mucosal barrier repair and controlling intestinal microbiota homeostasis, Zuogui Jiangtang Qinggan Prescription reduces intestinal mucosal damage induced by T2DM and NAFLD.

Effect of Yuehua Capsule on Autophagy of Macrophage Infected with Multi-drug Resistant Mycobacterium Tuberculosis / 中国实验方剂学杂志

Objective::This study aims to explore the effect and mechanism of Yuehua capsule serum for autophagy of macrophages infected with multi-drug resistant mycobacterium tuberculosis. Method::The rats were undertaken intragastric gavage with Yuehua capsule by 3.02 g·kg-1 once a day which was produced through low temperature condensation drying method. After 7 days, blood of abdominal aorta of rats was collected to prepare Yuehua capsule serum. RAW264.7 andmultidrug resistant tuberculosis were cultured in vitro.According to cell counting kit-8(CCK-8), 10% drug-containing serum was considered as the effective concentration. The cultured cells were divided into four groups: model groups(10% fetal bovine serum). Yuehua capsule serum(10% Yuehua capsule serum). Autophagy inhibitor group+ 3-MA+ Yuehua capsule medicated serum(3-MA+ 10% Yuehua capsule serum). Rapamycin (Rap) positive control group(200 mg·L-1 Rap+ 10% Yuehua capsule serum). Except for the normal group, the cells of each group were cultured for 24 h and infected for 4 h according to cell-bacteria 1∶10.Testing index: observation of autophagosomes under transmission electron microscope, the test of expression of microtubule-associated protein light chain-3Ⅱ(LC-3Ⅱ), microtubule-associated protein LC 3-Ⅱ/microtubule-associated protein light chain 3-Ⅰ(LC3-Ⅰ) and Beclin-1 with Western blot, indirect immunofluorescence staining for LC3B, and mRNA of Beclin-1 as well as LC3 with real-time fluorescent quantitative polymerase chain reaction(Real-time PCR). Result::Compared with normal group, model group did not see autophagy body cells, cells in the LC-3 Ⅱ, LC-3 Ⅱ/LC-3 Ⅰ, Beclin-1 protein and LC3, Beclin-1 mRNA gene expression level had no significant change, the cells without fluorescent particles, spots, no fluorescence intensity.Compared with model group, Yuehua capsules serum group and Rap positive control group can be observed the formation of phage, mRNA andprotein expression levelof LC-3 Ⅱ, LC-3 Ⅱ/LC-3 Ⅰ, Beclin-1 and LC3, Beclin-1 were significantly increased (P<0.05). Autophagy inhibitor group+ 3-MA+ Yuehua capsule medicated serum did not see autophagy, the mRNA and protein expression level of LC-3 Ⅱ, LC-3Ⅱ/LC-3Ⅰ, Beclin-1 and LC3, Beclin-1 were no significantly increased. Conclusion::Yuehua capsule medicated serum could induce autophagy of macrophages of RAW264.7.The mechanism was probably accomplished through regulating the expression level of autophagy key protein LC3, autophagosome mature protein Beclin-1 and relevant gene, meanwhile the conversion of LC3-I to LC3-Ⅱ was accelerated.