文献类型：期刊文献

中文题名：基于HAT/HDAC平衡调控PPARγ/PI3K/Akt通路活化探讨黄连化浊胶囊对糖尿病大血管病变的作用

英文题名：Effect of Huanglian Huazhuo capsule on diabetic macrovascular disease by regulating PPARγ/PI3K/Akt pathway based on HAT/HDAC balance

作者：王苑铭[1];朱瑾[1];杨维杰[1];易希善[2];肖国庆[2];康学东[1]

第一作者：王苑铭

机构：[1]甘肃中医药大学附属医院内分泌科,兰州730200;[2]甘肃中医药大学,兰州730000

第一机构：甘肃中医药大学第二附属医院

年份：2023

卷号：44

期号：1

起止页码：72

中文期刊名：海军军医大学学报

外文期刊名：Academic Journal of Naval Medical University

收录：CSTPCD;;Scopus;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：甘肃省中医药科研课题(GZKP-2020-30)。

语种：中文

中文关键词：糖尿病大血管病;黄连化浊胶囊;组蛋白乙酰转移酶;组蛋白脱乙酰酶;过氧化物酶体增殖物激活受体γ;磷脂酰肌醇3-激酶;蛋白激酶B

外文关键词：diabetic macrovascular disease;Huanglian Huazhuo capsule;histone acetyltransferase;histone deacetylase;peroxisome proliferator-activated receptorγ;phosphatidylinositol 3-kinase;protein kinase B

摘要：目的 探讨基于组蛋白乙酰转移酶(HAT)/组蛋白脱乙酰酶(HDAC)平衡调控过氧化物酶体增殖物激活受体γ(PPARγ)/PI3K/Akt通路活化探讨黄连化浊胶囊对糖尿病大血管病变的影响。方法 40只载脂蛋白E(ApoE)基因剔除小鼠均建立糖尿病大血管病变模型,随机分为模型组、阿托伐他汀组、黄连化浊胶囊组、阿托伐他汀+黄连化浊胶囊组,每组10只。10只野生型C57小鼠为空白组。阿托伐他汀组小鼠灌胃阿托伐他汀10 mg/kg,黄连化浊胶囊组小鼠灌胃黄连化浊胶囊0.675 g/kg,阿托伐他汀+黄连化浊胶囊组小鼠灌胃给予阿托伐他汀10 mg/kg+黄连化浊胶囊0.675 g/kg,空白组及模型组小鼠灌胃等体积生理盐水,每天1次,连续8周。培养小鼠胸主动脉平滑肌细胞,分为空白组、人氧化低密度脂蛋白(ox-LDL)组、ox-LDL+PPARγ沉默组、ox-LDL+PPARγ过表达组,除空白组采用等体积空白血清培养外,其他3组细胞均加入100μg/mL ox-LDL处理并采用含200μg/mL黄连化浊胶囊的血清共培养,ox-LDL+PPARγ沉默组进行PPARγ siRNA转染,ox-LDL+PPARγ过表达组转染PPARγ质粒。采用全自动生化分析仪检测小鼠血糖及血脂水平,H-E染色观察小鼠胸主动脉病理形态,流式细胞术检测平滑肌细胞凋亡率,蛋白质印迹法和qPCR分别检测细胞中HAT1、HDAC1、PPARγ、PI3K、Akt蛋白和mRNA的表达。结果 与模型组比较,阿托伐他汀组、黄连化浊胶囊组及阿托伐他汀+黄连化浊胶囊组小鼠餐后血糖、总胆固醇、甘油三酯及低密度脂蛋白胆固醇水平均降低,高密度脂蛋白胆固醇水平均升高(P均<0.05);与模型组相比,阿托伐他汀组、黄连化浊胶囊组小鼠主动脉血管内壁层次紊乱改善,可见少量的泡沫细胞,阿托伐他汀+黄连化浊胶囊组小鼠胸主动脉血管内壁层次紊乱改善优于阿托伐他汀组及黄连化浊胶囊组。ox-LDL+PPARγ沉默组平滑肌细胞凋亡率低于ox-LDL组和ox-LDL+PPARγ过表达组(P均<0.05);与ox-LDL组相比,ox-LDL+PPARγ沉默组平滑肌细胞中HAT1蛋白和mRNA表达均升高,而HDAC1、PPARγ、PI3K、Akt蛋白和mRNA表达均降低(P均<0.05);与ox-LDL+PPARγ沉默组相比,ox-LDL+PPARγ过表达组平滑肌细胞中HAT1蛋白和mRNA表达均降低,HDAC1、PPARγ、PI3K、Akt蛋白和mRNA表达均升高(P均<0.05)。结论 黄连化浊胶囊可能通过抑制HAT表达促进HDAC1表达和PPARγ/PI3K/Akt通路活化,抑制糖尿病大血管病变小鼠代谢紊乱,增加平滑肌细胞凋亡,减少动脉斑块形成,发挥保护作用。
Objective To investigate the effect of Huanglian Huazhuo capsule on diabetic macrovascular disease by regulating peroxisome proliferator-activated receptor γ(PPARγ)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akte) pathway based on histone acetyltransferase(HAT)/histone deacetylase(HDAC) balance. Methods Forty apolipoprotein E(ApoE) gene deleted mice were randomly divided into model group, atorvastatin group, Huanglian Huazhuo capsule group, and atorvastatin+Huanglian Huazhuo capsule group, with 10 in each group. Ten wild-type C57mice were included in the blank group. The mice in the atorvastatin group were gavaged with atorvastatin at a dose of 10 mg/kg, the mice in the Huanglian Huazhuo capsule group were gavaged with Huanglian Huazhuo capsule aqueous solution at a dose of 0.675 g/kg, the mice in the atorvastatin+Huanglian Huazhuo group were gavaged with atorvastatin at a dose of 10 mg/kg and Huanglian Huazhuo capsule aqueous solution at a dose of 0.675 g/kg, and the mice in the blank group and model group were gavaged with normal saline at the same dose, once daily for 8 weeks. Mouse thoracic aortic smooth muscle cells were collected and divided into blank group, human oxidized low density lipoprotein(ox-LDL) group,ox-LDL+PPARγ silent group, and ox-LDL+PPARγ overexpression group. The blank group was cultured with blank serum,and the other 3 groups were treated with 100 μg/mL ox-LDL, and co-cultured with serum containing 200 μg/mL Huanglian Huazhuo capsule. The ox-LDL+PPARγ silent group was transfected with PPARγ small interfering RNA, and ox-LDL+PPARγ overexpression group was transfected with PPARγ plasmid. Automatic biochemical analyzer was used to detect the blood glucose and blood lipid levels of mice, hematoxylin-eosin staining was used to observe the pathological morphology of mice thoracic aorta, flow cytometry was used to detect the apoptosis rate of smooth muscle cells, and Western blotting and quantitative ploymerase chain reaction were used to detect the protein and mRNA expression of HAT1, HDAC1, PPARγ, PI3K and Akt. Results Compared with the model group, the contents of postprandial blood glucose(PBG), total cholesterol(TC),triglyceride(TG) and low density lipoprotein-cholesterol(LDL-C) of mice in the atorvastatin group, Huanglian Huazhuo capsule group and atorvastatin+Huanglian Huazhuo capsule group were significantly decreased, while the contents of high density lipoprotein-cholesterol(HDL-C) were significantly increased(all P<0.05). Compared with the model group, the inner wall disorder was improved in the atorvastatin group and Huanglian Huazhuo capsule group, and a small number of foam cells were observed. The inner wall of thoracic aorta had the most significant improvement in the atorvastatin+Huanglian Huazhuo capsule group. The apoptosis rate of smooth muscle cells in the ox-LDL+PPARγ silent group was significantly lower than those in the ox-LDL group and ox-LDL+PPARγ overexpression group(both P<0.05). Compared with the ox-LDL group, the expression of HAT1 protein and mRNA in smooth muscle cells of the ox-LDL+PPARγ silent group was significantly increased, while the expression of HDAC1, PPARγ, PI3K and Akt protein and mRNA was significantly decreased(all P<0.05). Compared with the ox-LDL+PPARγ silent group, the expression of HAT1 protein and mRNA in smooth muscle cells of ox-LDL+PPARγ overexpression group was significantly decreased, while the expression of HDAC1,PPARγ, PI3K, Akt protein and mRNA was significantly increased(all P<0.05). Conclusion Huanglian Huazhuo capsule can improve the expression of HDAC1 and the activation of PPARγ/PI3K/Akt pathway by inhibiting the expression of HAT,inhibit the metabolic disorder of mice with diabetic macrovascular disease, increase the apoptosis of smooth muscle cells, and reduce the formation of arterial plaque, displaying a protective role.