1.Effects of Shizidaiping formula on MIN6 cell apoptosis and expressions of MEK1/2 and ERK1/2
Ruiting CHENG ; Yushan DONG ; Jian LI ; Xiaojin LA ; Chunyu TIAN ; Xiujuan GAO ; Xuemei ZHOU ; Qianru FU ; Bo WU ; Siqi DONG
Chinese Journal of Tissue Engineering Research 2017;21(4):603-608
BACKGROUND:Apoptosis of islet cel s is closely related to the long-term hyperglycemia-and hyperlipemia-induced injuries. OBJECTIVE:To observe the effect of Shizidaiping formula on the apoptosis and insulin secretion in MIN6 cel s under the high glucose and lipid environment, and to explore the protective effect of Shizidaiping formula and the related apoptosis mechanism. METHODS:MIN6 cel s were divided into normal, model, melbine, low-, medium-and high-dose Shizidaiping formula groups. The cel activity was examined by cel counting kit-8, the insulin secretion was measured by ELISA, the rate of apoptosis was measured by Annexin V-FITC&PI and the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 were examined by western blot assay. RESULTS AND CONCLUSION:Shizidaiping formula significantly improved MIN6 cel activity under high glucose and lipid condition (P<0.05), decreased early cel apoptosis, increased the level of insulin stimulated by low glucose in cel supernatant (P<0.05), and improved the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 (P<0.05). These results suggest that Shizidaiping formula can protect islet cel s from hyperglycemia and hyperlipemia damage by improving the activity of MIN6 cel s, reducing the insulin secretion and inhibiting the apoptosis of pancreaticβcel s in MIN6 cel s.
2.Mechanism of Zishen Qinggan Prescription in Improving Glucose and Lipid Metabolism in Type 2 Diabetes Based on Transcriptomics
Jingcun WANG ; Chunyu TIAN ; Fan ZHANG ; Xiaojin LA ; Fanwu WU ; Liang ZHU ; Leilei MA
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):109-117
ObjectiveBased on network pharmacology and transcriptomics, the mechanism of Zishen Qinggan prescription (ZSQGF) in improving glucose and lipid metabolism in type 2 diabetes (T2DM) model rats was explored. MethodBased on network pharmacology analysis of the differential genes between ZSQGF and T2DM, gene ontology(GO)analysis and Kyoto encyclopedia of genes and genomes(KEGG) analysis were conducted, and molecular docking analysis was used to verify the binding between components and targets. A T2DM rat model was established by high-fat feeding and injection of streptozotocin (STZ). The rats were randomly divided into the control group, model group, metformin (Met, 72 mg·kg-1) group, and ZSQGF high-, medium-, and low-dose groups (ZSQGF-H, ZSQGF-M, and ZSQGF-L, with 4.8, 2.4, and 1.2 g·kg-1 raw drug in the solution). The living status of rats was monitored and the levels of total cholesterol (TC), total triglycerides (TG), high density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in rat serum were detected. The liver tissues were subjected to Hematoxylin eosin(HE) staining and oil red O staining. The differential genes were analyzed through transcriptomics, GO and KEGG analysis, and the protein-protein interaction(PPI) network was obtained to screen key targets. With network pharmacology and transcriptomics analysis results, the protein pathways were identified. The expression levels of nuclear factor-κB (NF-κB), matrix metalloproteinase(MMP)-1 and MMP-9 proteins in liver tissues were detected by Western blot. The mRNA expression of B-cell lymphoma-2(Bcl-2) modifying factor(BMF), nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 4 (NOX4), and fatty acid synthase(FASN) was detected by real-time polymerase chain reaction(Real-time PCR). The expression of MMP-1 and MMP-9 in the liver was detected by immunofluorescence staining. ResultTranscriptomics and network pharmacology analysis suggested that ZSQGF may protect the liver through the glucose and lipid metabolism pathway and the inflammation pathway. Experiments showed that after 8 weeks of administration, the body weight, blood sugar, serum indicators, and pathological staining results of rats were improved. Western blot results indicated a decrease in the relative expression levels of NF-κB, MMP-1 and MMP-9 proteins in the liver. Real-time PCR results showed a decrease in the transcriptional expression of BMF, NOX4, and FASN in the ZSQGF-H group, while immunofluorescence staining results present decreased expression of MMP-1 and MMP-9 in the ZSQGF groups. ConclusionZSQGF can improve the glucose and lipid metabolism by inhibiting the expression of FASN, reducing lipid synthesis, and regulating the NF-κB signaling pathway.