摘要
目的探讨温阳利水方通过激活RhoA/ROCK通路改善慢性肺源性心脏病小鼠心肌细胞焦亡的机制。方法60只健康雄性C57BL/6小鼠用于实验研究。将小鼠暴露于烟雾和高压氧舱中诱导慢性肺源性心脏病(chronic pulmonary heart disease,CPHD)模型。将小鼠分为以下3组:对照组(正常条件下饲养的小鼠,接受清洁空气和0.9%氯化钠溶液作为模型组和温阳利水方组的对照,n=20),模型组(如前所述建立的CPF模型,n=20),温阳利水方组(在模型小鼠的基础上给予温阳利水方药剂治疗,n=20)。通过qPCR分析RhoA、ROCK1和ROCK2 mRNA表达。通过蛋白质印迹分析小鼠心肌组织p-MBS/t-MBS比值。通过酶联免疫吸附测定(ELISA)检测小鼠外周血炎症因子肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素1β(interleukin-1β,IL-1β)和白细胞介素6(interleukin-6,IL-6)水平。组织学分析小鼠心肌受损情况。通过超声心动图检测小鼠心肌功能。通过PI染色检测焦亡细胞。通过蛋白质印迹分析焦亡相关因子高迁移率族蛋白(HMGB1)和热休克蛋白70(HSP70)的表达。结果模型组RhoA、ROCK1和ROCK2 mRNA表达较对照组升高(P<0.05),温阳利水方组RhoA、ROCK1和ROCK2 mRNA表达较模型组降低(P<0.05)。模型组p-MBS/t-MBS比值较对照组升高(P<0.05),温阳利水方组p-MBS/t-MBS比值较模型组降低(P<0.05)。模型组TNF-α、IL-1β和IL-6水平较对照组升高(P<0.05),温阳利水方组TNF-α、IL-1β和IL-6水平较模型组降低(P<0.05)。HE染色显示,与对照组小鼠心肌相比,模型组小鼠心肌细胞排列紊乱,细胞肥大,炎症细胞浸润明显(P<0.05),与模型组相比,温阳利水方显著改善了CPHD引起的病理改变(P<0.05)。马松染色作为纤维化的病理检查表明,CPHD可导致小鼠严重的心肌纤维化,温阳利水方治疗可显著减少胶原沉积(P<0.05)。模型组小鼠左室收缩压(left ventricular systolic pressure,LVSP)、左室射血分数(left ventricular ejection fraction,LVEF)和心搏量(stroke volume,SV)较对照组降低(P<0.05),温阳利水方组LVSP、LVEF和SV较模型组升高(P<0.05)。模型组小鼠心肌细胞焦亡率较对照组升高(P<0.05),温阳利水方组小鼠心肌细胞焦亡率较模型组降低(P<0.05)。模型组HMGB1和HSP70的表达较对照组升高(P<0.05),温阳利水方组HMGB1和HSP70的蛋白表达较模型组降低(P<0.05)。结论温阳利水方通过激活RhoA/ROCK通路,显著改善了CPHD小鼠模型中心肌细胞焦亡和心肌结构及功能。
Objective To explore the mechanism of Wenyang Lishui Recipe(温阳利水方)improving cardiomyocyte pyroptosis in mice with chronic pulmonary heart disease by activating the RhoA/ROCK pathway.Methods Sixty healthy male C57BL/6 mice were used for the experimental studies.The chronic pulmonary heart disease(CPHD)model was induced in mice by exposing them to smoke and a hyperbaric oxygen chamber.The mice were divided into the following three groups:control group(mice raised under normal conditions,receiving clean air and normal saline as a control for the model group and Wenyang Lishui Recipe group,n=20),model group(as described previously established CPF model,n=20),Wenyang Lishui Recipe group(the model mice treated with Wenyang Lishui Recipe,n=20).RhoA,ROCK1 and ROCK2 mRNA expressions were analyzed by qPCR.The p-MBS/t-MBS ratio of the myocardial tissue was analyzed by Western blotting.The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in the peripheral blood of mice were detected by Enzyme-Linked Immunosorbent Assay(ELISA).Histological analysis was used to detect the myocardial damage in mice.Myocardial function in mice was measured by echocardiography method.Pyroptotic cells were detected by PI staining.The expressions of pyroptosis-related factors HMGB1 and HSP70 were analyzed by Western blotting.Results The expressions of RhoA,ROCK1 and ROCK2 mRNA in the model group were higher than those in the control group(P<0.05),and the expressions of RhoA,ROCK1 and ROCK2 mRNA in the Wenyang Lishui Recipe group were lower than those in the model group(P<0.05).The p-MBS/t-MBS ratio of the model group was higher than that of the control group(P<0.05),and the p-MBS/t-MBS ratio of the Wenyang Lishui Recipe group was lower than that of the model group(P<0.05).The levels of TNF-α,IL-1βand IL-6 in the model group were higher than those in the control group(P<0.05),while the levels of TNF-α,IL-1βand IL-6 in the Wenyang Lishui Recipe group were lower than those in the model group(P<0.05).HE staining showed that compared with the myocardium of mice in the control group,the cardiomyocytes of the mice in the model group were arranged disorderly,the cells were hypertrophic,and the infiltration of inflammatory cells was obvious(P<0.05).Compared with the model group,Wenyang Lishui Recipe significantly improved pathological changes caused by CPHD(P<0.05).Masson staining as a pathological examination of fibrosis showed that CPHD can cause severe myocardial fibrosis in mice,and the treatment with Wenyang Lishui Recipe can significantly reduce collagen deposition(P<0.05).The values of left ventricular systolic pressure(LVSP),left ventricular ejection fraction(LVEF)and stroke volume(SV)of mice in the model group were lower than those in the control group(P<0.05),while those in the Wenyang Lishui Recipe group were higher than those in the model group(P<0.05).The pyroptosis rate of myocardial cells in mice in the model group was higher than that in the control group(P<0.05),and the pyroptosis rate of myocardial cells in the Wenyang Lishui Recipe group was lower than that in the model group(P<0.05).The expressions of HMGB1 and HSP70 in the model group were higher than those in the control group(P<0.05),and the protein expressions of HMGB1 and HSP70 in the Wenyang Lishui Recipe group were lower than those in the model group(P<0.05).Conclusion Wenyang Lishui Recipe activates RhoA/ROCK pathway,significantly improving cardiomyocyte pyroptosis,myocardial structure and function in the CPHD mouse model.
作者
陈昕芳
李红
苏士成
吴峰妹
黄辉
CHEN Xinfang;LI Hong;SU Shicheng;WU Fengmei;HUANG Hui(Kunshan Traditional Chinese Medicine Hospital,Kunshan 215300,Jiangsu,China)
出处
《辽宁中医杂志》
北大核心
2026年第2期204-208,F0003,共6页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金青年科学基金项目(82004271)
苏州市姑苏卫生人才科研项目(GSWS2022107)。
