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.Effect of apoptosis inhibitor of macrophage in inflammatory reactions and lipid metabolic diseases
Fan ZHANG ; Chunyu TIAN ; Jingcun WANG ; Xiaojin LA ; Qianru FU ; Jie LI ; Wenhao FU
Chinese Journal of Comparative Medicine 2024;34(3):142-148
Apoptosis inhibitor of macrophage(AIM)belongs to group B of the scavenger receptor cysteine rich-super family.AIM is a soluble protein secreted by macrophages.The expression of this protein is controlled by the liver X receptor.AIM,which is secreted by macrophages,plays important and broad roles in the immune responses of the body.It not only inhibits the apoptosis of macrophages but also participates in the regulation of macrophage polarization.In addition,studies have revealed that AIM is involved in various physiological and pathological processes,such as inflammation,obesity,atherosclerosis,and cancer.It has been used as a biological marker for the diagnosis of diseases such as tuberculosis and liver cirrhosis.Moreover,it can promote the lipolysis of adipose cells by inhibiting the activity of fatty acid synthase(FAS),playing an important role in the regulation of lipid homeostasis,lipid metabolism,and autoimmune diseases.In this paper,we review the multiple functional characteristics of AIM and its effects on inflammation,lipid metabolism,and related diseases to provide a theoretical basis for relevant medical research.
3.Regulation of Suaeda salsa in renal AIM and macrophage polarization in rats with diabetes kidney disease
Fan ZHANG ; Chunyu TIAN ; Jingcun WANG ; Xiaojin LA ; Liang ZHU ; Hong CHANG ; Biwei ZHANG ; Ji'an LI
International Journal of Traditional Chinese Medicine 2024;46(9):1155-1162
Objective:To explore the regulatory effect of Suaeda salsa on renal apoprosis inhibitor of macrophage (AIM) and macrophage polarization in diabetes kidney disease (DKD) model rats.Methods:A DKD rat model was established using a high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin (STZ). The rats were divided into model group, metformin group, and Suaeda salsa high-, medium-, and low-dosage groups using a random number table method, with 8 mice in each group. The normal group was set with 8 rats in this group. The metformin group was given 85.71 mg/kg of metformin solution by gavage, while the Suaeda salsa high-, medium-, and low-dosage groups were given 3.08, 1.54, and 0.77 g/kg of Suaeda salsa suspension by gavage (raw dosage). The normal group and model group were given equal volumes of saline by gavage, once a day, administered by gavage for 12 weeks of intervention. Fasting blood glucose (FBG) and glucose tolerance (OGTT) were measured, and the area under the curve (AUC) was calculated; the levels of glycosylated hemoglobin (HbA1c) and glycosylated serum protein (GSP) were detected; urea nitrogen (BUN), serum creatinine (SCr), and 24-hour urine protein (24 hUP) were detected; HE staining was used to observe the pathological morphology of the kidneys; Masson and PAS staining were used to observe renal tissue fibrosis; Western blot method was used for detecting AIM, CD206, CD86, TNF-α and IL-10 protein levels in renal tissue; Immunofluorescence was used to detect the average optical density values of AIM, CD206, and CD86 proteins in renal tissue.Results:Compared with the model group, the FBG, OGTT AUC, HbA1c, GSP of each dosage group of Suaeda salsa decreased ( P<0.01); the expression levels of AIM, CD206, and IL-10 proteins in renal tissue increased ( P<0.01 or P<0.05), while the expression levels of CD86 and TNF-α protein significantly decreased ( P<0.01 or P<0.05); HE, Masson, and PAS staining results showed that compared with the model group, the changes in renal microvasculature and renal fibrosis of rats in each dosage group of Suaeda salsa were improved. Conclusion:Suaeda salsa may regulate AIM, promote polarization of M2 macrophages, improve the inflammatory microenvironment of macrophages, thereby lowering blood lipids of DKD rats, and improving renal pathological damage.
4.Exploration on the effects of Zhiganqing Prescription on insulin resistance and gluconeogenesis in NAFLD mice based on the PI3K/Akt/FoxO1 signaling pathway
Mingyu SHEN ; Jingya WANG ; Wenxuan XU ; Shuo WANG ; Leilei MA ; Jiahao HU ; Chao LI ; Xiaojin LA ; Ji'an LI
International Journal of Traditional Chinese Medicine 2024;46(10):1295-1302
Objective:To observe the protective effects of Zhiganqing Prescription on the liver of C57BL/6J non-alcoholic fatty liver disease (NAFLD) mice induced by high fat diet and its effects on PI3K/Akt/FoxO1 signaling pathway, insulin resistance (IR) and gluconogenesis.Methods:A total of 48 8-week-old male C57BL/6J mice were divided into control group ( n=8) and modeling group ( n=40) according to random number table method. The control group was fed with ordinary diet, and the model group was fed with high-fat diet. The NAFLD model was established after 8 weeks of feeding. The modeling group was divided into model group, Pioglitazone group, Zhiganqing Prescription low-, medium-, and high-dosage group ( n=8 in each group) according to random number table method, and drug intervention lasted for 8 weeks. The body mass of mice was measured regularly during administration. Fasting blood glucose (FBG) levels were measured at 0 and 8 weeks of administration, and oral glucose tolerance tests (OGTT) were conducted. After the experiment, serum levels of GPT, GOT, TC, TG, LDL-C, HDL-C, FINS and C-P were detected and HOMA-IR was calculated. The pathological morphology of liver was observed by HE and PAS staining. The expression levels of PI3K and p-Akt were detected by IHC staining. The protein expression levels of PI3K, Akt, p-Akt, FoxO1, p-FoxO1, G6PC and PCK1 were detected by Western blot. Results:Compared with model group, the body weight of mice in each administration group decreased at 4, 6 and 8 weeks ( P<0.05 or P<0.01). At the 8th week of administration, the levels of FBG and OGTT AUC in each administration group decreased ( P<0.05 or P<0.01), the levels of GPT, TC, TG and LDL-C decreased ( P<0.01), and the GOT levels in Zhiganqing Prescription medium- and high-dosage groups decreased ( P<0.01). The HDL-C level in Zhiganqing Prescription medium-dosage group decreased ( P<0.05 or P<0.01), and the HOMA-IR level in Zhiganqing Prescription low- and medium-dosage groups decreased ( P<0.05 or P<0.01). The levels of FINS and C-P in each administration group increased ( P<0.05 or P<0.01), and the expressions of PI3K protein and p-Akt/Akt, p-FoxO1 /FoxO1 protein in liver tissues increased ( P<0.05 or P<0.01). The protein expressions of G6PC and PCK1 decreased ( P<0.05 or P<0.01). Conclusion:Zhiganqing Prescription can effectively control the body mass, blood glucose, liver function and blood lipids of NAFLD mice, improve IR and gluconeogenesis, the mechanism of which may be related to the activation of PI3K/Akt/FoxO1 signaling pathway.
5.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.