ABSTRACT
This study aims to explore the effect of Zuogui Jiangtang Qinggan Formula(ZGJTQGF) on the lipid metabolism in the db/db mouse model of type 2 diabetes mellitus(T2DM) complicated with non-alcoholic fatty liver disease(NAFLD) via the insulin receptor(INSR)/adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)/sterol-regulatory element-binding protein 2(SREBP-2) signaling pathway. Twenty-four db/db mice were randomized into positive drug(metformin, 0.067 g·kg~(-1)) and low-(7.5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) ZGJTQGF groups. Six C57 mice were used as the blank group and administrated with an equal volume of distilled water. The mice in other groups except the blank group were administrated with corresponding drugs by gavage for 6 consecutive weeks. At the end of drug administration, fasting blood glucose(FBG) and blood lipid levels were measured, and oral glucose tolerance test was performed. Compared with the blank group, the mice treated with ZGJTQGF showed decreased body mass and liver weight coefficient, lowered levels of FBG, total cholesterol(TC), triglyceride(TG), and low-density lipoprotein(LDL), and weakened liver function. The pathological changes and lipid accumulation in the liver tissue were examined. Western blot was employed to measure the protein levels of INSR, AMPK, p-AMPK, and SREBP-2. Compared with the blank group, the model group showed down-regulated protein levels of INSR and p-AMPK/AMPK and up-regulated protein level of SREBP-2. Compared with the model group, high-dose ZGJTQGF up-regulated the protein levels of INSR and p-AMPK/AMPK and down-regulated the protein level of SREBP-2. Low-dose ZGJTQGF slightly up-regulated the protein levels of INSR and p-AMPK/AMPK and down-regulated the protein level of SREBP-2, without significant differences. The results suggested that ZGJTQGF may alleviate insulin resistance and improve lipid metabolism in db/db mice by activating the INSR/AMPK/SREBP-2 signaling pathway.
Subject(s)
Diabetes Mellitus, Type 2 , Drugs, Chinese Herbal , Non-alcoholic Fatty Liver Disease , Mice , Animals , Non-alcoholic Fatty Liver Disease/drug therapy , Non-alcoholic Fatty Liver Disease/metabolism , Diabetes Mellitus, Type 2/complications , Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/metabolism , Lipid Metabolism , AMP-Activated Protein Kinases/metabolism , Sterol Regulatory Element Binding Protein 1/metabolism , Liver , LipidsABSTRACT
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.
Subject(s)
Diabetes Mellitus, Type 2 , Gastrointestinal Microbiome , Non-alcoholic Fatty Liver Disease , Mice , Animals , Non-alcoholic Fatty Liver Disease/drug therapy , Non-alcoholic Fatty Liver Disease/metabolism , RNA, Ribosomal, 16S , Diabetes Mellitus, Type 2/drug therapy , Diabetes Mellitus, Type 2/metabolism , Occludin/metabolism , Occludin/pharmacology , Claudin-1/metabolism , Intestinal Mucosa , Liver , Triglycerides/metabolism , Diet, High-Fat , Homeostasis , Mice, Inbred C57BLABSTRACT
OBJECTIVE: To investigate the effect of Jiangtang Yishen Recipe (JTYSR) on high insulin induced cell proliferation of human glomerular mesangial cells (HMCs) and the expression of insulin receptor substrate 1 (IRS-1) and phosphatidylinositol-3-kinase (PI-3K). METHODS: HMCs were divided into 4 groups, i.e., the negative control group, the high insulin model group, the JTYSR group, and the LY294002 group. The concentration of insulin, JTYSR, and LY294002 was respectively confirmed by pre-experiment. Different culture solution was respectively added for different groups. RPMI1640 culture solution was added for HMCs in the negative control group, while HMCs in the rest 3 groups were cultured by 100 nmol/L insulin for 24 h. Meanwhile, HMCs from the JTYSR group and the LY294002 group were exposed to 125 mg/L JTYSR and 80 micromol/L LY294002 respectively for further 48 h. The proliferation of HMCs was detected by MTT and flow cytometry. The protein expression of IRS-1 and PI-3K in HMC was detected by immunohistochemical assay and Western blot. Results The proliferation of HMCs induced by high insulin could be significantly lowered, and the protein expression of IRS-1 and PI-3K could be down-regulated in the JTYSR group and the LY294002 group (P <0.01). Compared with the LY294002 group, the protein expression of IRS-1 and PI-3K could be slightly down-regulated in the JTYSR group (P <0.05). CONCLUSION: JTYSR could lower high insulin induced proliferation of HMCs, and its mechanism might be related to insulin signaling pathway.
Subject(s)
Cell Proliferation/drug effects , Drugs, Chinese Herbal/pharmacology , Insulin Receptor Substrate Proteins/metabolism , Mesangial Cells/physiology , Phosphatidylinositol 3-Kinases/metabolism , Chromones , Humans , Morpholines , Phosphatidylinositol 3-Kinase/metabolism , Signal TransductionABSTRACT
Flow injection chemiluminescence analysis was used to determine the antioxidation activity of extract from Morinda officinalis How. The determination was based on the inhibition effect of Morinda officinalis How extraction in the chemiluminescence reaction of luminol - H2O2-CuSO4 system, and vitamin C was used as positive control. It was showed that Morinda officinalis How original liquid has obvious anti-radical activity. Spectroscopy was applied to determine the inhibition ratio of Morinda officinalis How extraction for the superoxide anion and hydroxy radicals in the special system. It was shown that the extraction from Morinda officinalis How could obviously scavenge superoxide anion and hydroxy radicals. It is suggested that the Chinese Medicine Morinda Officinalis How is a potential antioxidation activity medicine.
Subject(s)
Antioxidants/chemistry , Luminescence , Morinda/chemistry , Antioxidants/pharmacology , Ascorbic Acid/chemistry , Ascorbic Acid/pharmacology , Copper Sulfate/chemistry , Flow Injection Analysis , Hydrogen Peroxide/chemistry , Hydroxyl Radical/antagonists & inhibitors , Hydroxyl Radical/chemistry , Luminescent Measurements/instrumentation , Luminescent Measurements/methods , Luminol/chemistry , Oxidation-Reduction/drug effects , Plant Extracts/analysis , Plant Extracts/chemistry , Spectrophotometry/instrumentation , Spectrophotometry/methods , Superoxides/antagonists & inhibitors , Superoxides/chemistryABSTRACT
Polysaccharide was extracted from morinda officinalis how by back-flowing with 80% ethanol for 1 h, and the method of improved phenol-sulfuric acid spectroscopy was adopted to determine the content of morinda officinalis how polysaccharide. As for its trace elements, the atomic absorption spectroscopy was used to detect the content of Zn, Fe and Cu, which were compared with its fake; and cold atomic fluorometry was applied to determine the trace mercury. The results were satisfactory, which can give reference about the effective components of morinda officinalis how, and will help to exploit it.