摘要
目的 研究基于Toll样受体4(TLR4)/核转录因子-κB(NF-κB)/NOD样受体蛋白3(NLRP3)信号通路探讨黄连解毒汤对冠状动脉粥样硬化性心脏病(CHD)小鼠心肌保护作用.方法 50只6周的雄性SD小鼠中取10只作正常对照组,其余小鼠给予高脂饮食建立CHD模型.采用随机数字表法将40只模型小鼠分为模型组、黄连解毒汤低、中、高剂量组,每组各10只.黄连解毒汤低、中、高剂量组每日分别以10、20、40 g/kg剂量进行灌胃,每日1次,持续6周;正常组、模型组每日给予等量蒸馏水灌胃.干预后采用生化法检测各组总胆固醇、甘油三酯、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)水平,酶联免疫吸附试验检测超氧化物歧化酶(SOD)、丙二醛、内皮素水平;采用苏木精-伊红染色法观察小鼠心肌细胞病理形态,蛋白质印迹法检测心肌组织中TLR4、NF-κB、NLRP3的表达.比较各组小鼠血脂、心肌组织中抗氧化相关指标和TLR4、NF-κB、NLRP3的表达.结果 模型组、黄连解毒汤低、中、高剂量组血脂水平均高于对照组,差异均有统计学意义(P＜0.05);黄连解毒汤低剂量组与模型组血脂水平比较,差异无统计学意义(P＞0.05);黄连解毒汤中、高剂量组总胆固醇、甘油三酯、LDL水平均低于模型组,HDL水平高于模型组,差异均有统计学意义(P＜0.05).模型组、黄连解毒汤低、中、高剂量组心肌组织中SOD水平均低于对照组,丙二醛、内皮素水平均高于对照组,差异均有统计学意义(P＜0.05);黄连解毒汤低、中、高剂量组SOD水平均高于模型组,丙二醛、内皮素水平均低于模型组,差异均有统计学意义(P＜0.05).对照组心肌纤维排列整齐,横纹清晰且细胞完整,无细胞肿胀、炎症浸润等;模型组心肌纤维排列紊乱,结构发生异常,有严重细胞肿胀、炎症浸润;黄连解毒汤低、中、高剂量组未见心肌纤维断裂,黄连解毒汤中、高剂量组细胞肿胀和炎症浸润情况优于黄连解毒汤低剂量组,黄连解毒汤高剂量组优于黄连解毒汤中剂量组.模型组、黄连解毒汤中、高剂量组TLR4、NF-κB、NLRP3蛋白表达量均高于对照组,差异均有统计学意义(P＜0.05);黄连解毒汤低、中、高剂量组TLR4、NF-κB、NLRP3蛋白表达量均低于模型组,差异均有统计学意义(P＜0.05).且黄连解毒汤各剂量组中以上指标变化呈剂量依赖性.结论 黄连解毒汤能降低CHD小鼠血脂水平,保护心肌细胞,其能够通过下调组织TLR4、NF-κB、NLRP3蛋白表达以调节血脂水平并改善心肌功能,且黄连解毒汤的效果随着剂量的增加而增强.
Abstract
Objective To investigate myocardial protective effect of Huanglian Jiedu Decoction on mice with coronary heart disease based on toll-like receptor 4/nuclear factor kappa B/NOD-like receptor family,pyrin domain containing 3(TLR4/NF-κB/NLRP3)signaling path-way.Methods Ten of 50 six-week-old male SD mice were set as normal controls,and the rest of the mice were fed with a high-fat diet to es-tablish CHD models.Forty CHD model mice were divided into model group,Huanglian Jiedu Decoction low-dose,medium-dose and high-dose groups according to the random number table method,with 10 mice in each group.The low,medium and high dose groups were administered by gastric gavage at a dose of 10,20 and 40 g/kg once daily for 6 weeks,respectively.In control group and model group,equal amounts of dis-tilled water were administered daily by gavage.Lipid profiles including total cholesterol,triglycerides,low-density lipoprotein(LDL)and high-density lipoprotein(HDL)in each group were measured biochemically.Malondialdehyde and superoxide dismutase(SOD)activity and endothelin level were detected by enzyme-linked immunosorbent assay(ELISA).The pathological morphology changes of myocardial tissue were observed by hematoxylin-eosin staining.And the protein expressions of TLR4,NF-κB and NLRP3 in myocardial tissue were detected by western blot-ting.Blood lipids,antioxidant related indicators in myocardial tissue,and TLR4,NF-κB,and NLRP3 expression were compared in each group of mice.Results The levels of blood lipid in the model group and the Huanglian Jiedu Decoction low,medium,and high-dose groups were higher than those in the control group,the differences were statistically significant(P＜0.05);there were no statistically significant differences in blood lipid levels between the Huanglian Jiedu Decoction low-dose group and the model group(P＞0.05);the levels of total cholesterol,tri-glyceride and LDL in the Huanglian Jiedu Decoction middle and high dose groups were lower than those in the model group,while the level of HDL was higher than that in the model group,the differences were statistically significant(P＜0.05).The levels of SOD in the myocardial tissue of the model group and the Huanglian Jiedu Decoction low,medium,and high-dose groups were lower than those of the control group,while the lev-els of malondialdehyde and endothelin were higher than those of the control group,and the differences were statistically significant(P＜0.05);the levels of SOD in the Huanglian Jiedu Decoction low,medium,and high-dose groups were higher than those in the model group,while the average levels of malondialdehyde and endothelin were lower than those in the model group,and the differences were statistically significant(P＜0.05).In the control group,myocardial fibers were arranged neatly,with clear transverse lines and intact cells,without cell swelling and inflammatory in-filtration.In the model group,the myocardial fibers were disordered,and the structure was abnormal,with severe cell swelling and inflammatory infiltration.No myocardial fiber rupture was found in each dose group of Huanglian Jiedu decoction.The cell swelling and inflammatory infiltration in the Huanglian Jiedu Decoction medium,and high-dose groups were better than those in the Huanglian Jiedu Decoction low dose group,and the Huanglian Jiedu Decoction high dose group was better than that in the Huanglian Jiedu Decoction medium dose group.The levels of TLR4,NF-κB,NLRP3 protein expression in the model group and the Huanglian Jiedu Decoction low,medium,and high-dose groups were higher than those in the control group,the differences were statistically significant(P＜0.05);the levels of TLR4,NF-κB,NLRP3 protein expression in the Huanglian Jiedu Decoction low,middle and high dose groups were lower than those in the model group,the differences were statistically significant(P＜0.05).The changes in the above indicators in each dose group of Huanglian Jiedu Tang were dose-dependent.Conclusion Huanglian Jiedu Decoction can reduce the blood lipid level and protect cardiomyocytes in CHD mice,which may be through the down-regulation of tissue TLR4,NF-κB,NLRP3 protein expression to regulate blood lipid levels and improve myocardial function,and the effect of Huanglian Jiedu De-coction increased with increasing dose.
基金项目
安徽省科研编制计划(2022)(2022AH050473)
维普
万方数据