基于GRP78/CHOP通路探讨补中益气汤对心衰大鼠心肌细胞内质网应激凋亡机制的影响  

Effect of Buzhong Yiqi Decoction(补中益气汤)on Mechanism of Endoplasmic Reticulum Stress Induced Apoptosis in Cardiac Myocytes of Heart Failure Rats Based on GRP78/CHOP pathway

作　　者：邓成康 魏东升 李计龙 郭宋丹 郑娴[1,2] DENG Chengkang;WEI Dongsheng;LI Jilong;GUO Songdan;ZHENG Xian(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China;Key Laboratory of The Theory and Application of Traditional Chinese Medicine of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学附属医院,辽宁沈阳110032 [3]辽宁中医药大学中医脏象理论及应用教育部重点实验室,辽宁沈阳110847

出　　处：《实用中医内科杂志》2024年第12期80-84,I0029-I0031,共8页Journal of Practical Traditional Chinese Internal Medicine

基　　金：辽宁省自然科学基金面上项目(2020-MS-227);辽宁中医药大学自然科学类重点项目(2021LZY053)。

摘　　要：目的研究旨在深入探讨补中益气汤对心衰大鼠心肌内质网应激凋亡机制的影响。方法研究将48只SD大鼠分为两组:空白组(12只)和造模组(36只)。空白组作为假手术组,而造模组则通过腹主动脉缩窄术建立心衰(HF)模型。48只SD大鼠进行分组,包括空白组12只与造模组36只。各组采用腹主动脉缩窄术法建立HF模型。经腹主动脉结扎后造模成功36只,将其分为模型组、盐酸曲美他嗪组(西药组)和补中益气汤组,连续4周内接受相应剂量的药物灌胃。通过ELISA检测血清NT-ProBNP的浓度。通过HE染色、Masson染色评价补中益气汤对心肌组织结构改变和纤维化程度的影响。通过采用化学荧光法检测心衰大鼠心肌组织活性氧(ROS)的含量。通过TUNEL染色评价心肌细胞凋亡的程度。通过Western blot(WB)检测内质网应激相关因子CHOP、GRP78、IRE1α、XBP1、PECK、ATF6和PUMA的蛋白表达。结果与空白组相比,模型组的血清NT-ProBNP值显著高于空白组(P<0.01),与模型组对比,补中益气汤组和西药组的NT-ProBNP值均呈现出下降的趋势。与模型组相比,补中益气汤组与西药组的心肌组织排列较整齐,形态较规则,炎症浸润较少,水肿减轻,纤维组织变少,间隙变窄。相较于空白组,模型组大鼠心肌组织中的ROS含量显著升高,相较于模型组,补中益气汤和西药组大鼠心肌组织中的ROS含量则显著下降。相较于空白组,模型组的心肌细胞在TUNEL染色后呈现出更高的阳性细胞比例(P<0.01);补中益气汤组及西药组中各指标变化不大,相对于模型组而言,补中益气汤组和西药组的心肌细胞TUNEL染色阳性细胞率呈现显著下降(P<0.01)。相较于空白组,模型组的CHOP、GRP78、IRE1α、XBP1、PECK、ATF6和PUMA的蛋白表达呈现显著上升的趋势(P<0.01);相对于模型组,补中益气汤组和西药组的CHOP、GRP78、IRE1α、XBP1、PECK、ATF6和PUMA的蛋白表达呈现下降趋势。结论�Objective To investigate the effect of Buzhong Yiqi Decoction(补中益气汤)on the mechanism of endoplasmic re⁃ticulum stress-induced apoptosis in heart failure rats.Methods Forty-eight SD rats were grouped,including blank group(12 rats)and model group(36 rats).The blank group was the sham operation group.The model of heart failure(HF)was estab⁃lished through abdominal aortic constriction surgery in the model group.After successful modeling through abdominal aortic liga⁃tion,36 rats were divided into model group,trimetazidine hydrochloride group(western medicine group)and Buzhong Yiqi Decoc⁃tion group,and the rats in each group received the corresponding doses of medication by gavage for 4 consecutive weeks.The con⁃centration of NT-ProBNP in serum was detected by ELISA.The effect of Buzhong Yiqi Decoction on changes in myocardial tis⁃sue structure and fibrosis degree was evaluated through HE staining and Masson staining.By using chemical fluorescence method,the content of reactive oxygen species(ROS)in myocardial tissue of heart failure was detected.The degree of myocardial cell apoptosis was evaluated through TUNEL staining.Western blot(WB)was used to detect the protein expressions of endoplasmic reticulum stress-related factors,including CHOP,GRP78,IRE1α,XBP1,PECK,ATF6 and PUMA.Results Compared with that in the blank group,the serum NT-ProBNP value in the model group was significantly higher(P<0.01).Compared with that in the model group,the NT-ProBNP value in the Buzhong Yiqi Decoction group and the western medicine group showed a down⁃ward trend.Compared with that of the model group,the myocardial tissue of the Buzhong Yiqi Decoction group and the western medicine group was more orderly,with regular morphology,less inflammatory infiltration,reduced edema,less fibrous tissue and narrower space.Compared with that of the blank group,the ROS content in the myocardial tissue of the model group was signifi⁃cantly increased,and compared with that of the model group,the ROS contents in the myocardia

关 键 词：心衰 心功能 内质网应激 细胞凋亡 补中益气汤 

分 类 号：R285.5[医药卫生—中药学]