文献类型：期刊文献

中文题名：黄芪甲苷通过调控GPR35/AMPK/mTOR信号轴改善线粒体能量代谢抑制放射性大鼠心肌纤维化

英文题名：AstragalosideⅣameliorates mitochondrial energy metabolism and inhibits radiation-induced myocardial fibrosis in rats via regulating GPR35/AMPK/mTOR signaling axis

作者：任春贞[1,2,4];李倩蓉[1,2];舒畅[1,2,3];陈其林[1,2];李琳婵[1,2];蒋虎刚[1,2,4];支晓东[1,2,4];赵信科[1,2,4];李应东[1,2,4]

第一作者：任春贞

机构：[1]甘肃中医药大学中西医结合学院,甘肃兰州730000;[2]甘肃省中医药防治慢性病重点实验室,甘肃兰州730000;[3]甘肃中医药大学国际教育学院,甘肃兰州730000;[4]甘肃中医药大学附属医院,甘肃兰州730000

第一机构：甘肃中医药大学中西医结合学院

年份：2026

卷号：51

期号：2

起止页码：505

中文期刊名：中国中药杂志

外文期刊名：China Journal of Chinese Materia Medica

收录：;北大核心:【北大核心2023】；

基金：国家自然科学基金项目(82360926,82374279);中医药传承与创新“百千万”人才工程(岐黄工程)青年岐黄学者基金项目;国家中医药管理局“李应东全国名中医传承工作室”建设项目(国中医药办人教函[2022]5号);甘肃省科技重大专项计划项目(20ZD7FA002);“双一流”科研重点项目(GSSYLXM-05);甘肃教育揭榜挂帅项目(2021jyjbgs-03);2024年甘肃省优秀博士生项目(24JRRA569)。

语种：中文

中文关键词：黄芪甲苷;放射性心肌纤维化;线粒体功能;能量代谢;GPR35/AMPK/mTOR信号轴

外文关键词：astragalosideⅣ;radiation-induced myocardial fibrosis;mitochondrial function;energy metabolism;GPR35/AMPK/mTOR signaling axis

摘要：旨在探讨G蛋白偶联受体35(GPR35)/AMP活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)信号轴介导的线粒体能量代谢在放射性心肌纤维化中的作用机制及黄芪甲苷的干预作用。将40只Wistar大鼠随机分为4组,每组10只。除空白组不予辐射外,其余各组大鼠均经麻醉后接受40 Gy X射线单次局部胸部照射1次,建立放射性心肌纤维化大鼠模型。辐射后,黄芪甲苷各干预组以80、160 mg·kg^(-1)剂量分别灌胃,每日1次,连续给药30 d。连续给药30 d后,ELISA法检测血清氨基末端BNP前体(NT-proBNP)、心肌肌钙蛋白I(cTnI)、肌酸激酶同工酶(CK-MB)含量;苏木素-伊红(HE)、Masson染色观察心肌组织病理变化;TUNEL染色检测心肌细胞凋亡情况;比色法检测心肌能量代谢腺嘌呤核苷三磷酸(ATP)、腺嘌呤核苷二磷酸(ADP)、腺嘌呤核苷一磷酸(AMP)水平及心肌线粒体呼吸链复合物Ⅰ~Ⅳ活性;荧光探针检测心肌组织Ca^(2+)荧光强度表达;透射电镜检测心肌超微结构;免疫荧光及Western blot检测心肌组织GPR35、磷酸化AMPK(p-AMPK)、磷酸化mTOR(p-mTOR)、胱天蛋白酶-3(Caspase-3)、胱天蛋白酶-9(Caspase-9)、B细胞淋巴瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、Ⅰ型胶原蛋白(COL-Ⅰ)、α-平滑肌肌动蛋白(α-SMA)荧光及蛋白表达。结果显示,与空白组相比,模型组大鼠血清NT-proBNP、cTnI、CK-MB含量明显升高;心肌组织可见局灶性心肌细胞坏死溶解,结缔组织增生,胶原纤维阳性面积增多;心肌细胞凋亡率升高;ATP水平显著下降,AMP和ADP水平明显升高,线粒体呼吸链复合物Ⅰ~Ⅳ活性明显降低;心肌细胞Ca^(2+)荧光显著升高;心肌线粒体破损严重,呈絮状改变,受损线粒体增多、肿胀、破裂、排列紊乱,嵴紊乱,部分线粒体出现空泡。心肌组织GPR35、p-AMPK、Bax、Caspase-3、Caspase-9、COL-Ⅰ、α-SMA蛋白及荧光表达明显升高,p-mTOR、Bcl-2蛋白及荧光表达下降。与模型组相比,黄芪甲苷可明显抑制NT-proBNP、cTnI、CK-MB含量;心肌细胞凋亡率显著下降;ATP水平显著升高,AMP和ADP水平明显降低,线粒体呼吸链复合物Ⅰ~Ⅳ活性明显升高;Ca^(2+)荧光强度降低;线粒体损伤缓解,排列较整齐;GPR35、p-AMPK、Bax、Caspase-3、Caspase-9、COL-Ⅰ、α-SMA蛋白及荧光表达明显下降,p-mTOR、Bcl-2蛋白及荧光表达升高。综上所述,黄芪甲苷对放射性心肌纤维化具有抑制作用,其机制可能是通过调控GPR35/AMPK/mTOR信号轴改善线粒体能量代谢发挥作用。
This study aimed to elucidate the role of the G protein-coupled receptor 35(GPR35)/AMP-activated protein kinase(AMPK)/mechanistic target of rapamycin(mTOR)signaling axis in mediating mitochondrial energy metabolism in radiation-induced myocardial fibrosis(RIMF)and to evaluate the interventional effect of astragalosideⅣ(AS-Ⅳ).Forty Wistar rats were randomly divided into four groups(n=10 per group).Except for the blank group,all rats were anesthetized and subjected to a single 40-Gy local thoracic X-ray irradiation to establish the RIMF model.After irradiation,AS-Ⅳwas administered intragastrically once daily at doses of 80 and 160 mg·kg^(-1) for 30 consecutive days.After 30 days of continuous administration,the serum levels of N-terminal pro-B-type natriuretic peptide(NT-proBNP),cardiac troponinI(cTnI),and creatine kinase-MB(CK-MB)were quantified by ELISA.Hematoxylin-eosin(HE)and Masson staining were used to observe myocardial histopathological changes,and TUNEL staining was performed to detect cardiomyocyte apoptosis.The levels of adenosine triphosphate(ATP),adenosine diphosphate(ADP),and adenosine monophosphate(AMP),as well as the activities of mitochondrial respiratory chain complexesⅠ-Ⅳ,were measured colorimetrically.The fluorescence intensity of Ca^(2+)in myocardial tissue was detected using a fluorescent probe,and ultrastructural changes were observed by transmission electron microscopy.The fluorescence intensity and protein expression levels of GPR35,phosphorylated AMPK(p-AMPK),phosphorylated mTOR(p-mTOR),Caspase-3,Caspase-9,B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),typeⅠcollagen(COL-Ⅰ),andα-smooth muscle actin(α-SMA)in myocardial tissue were analyzed by immunofluorescence and Western blot.Results showed that compared with the blank group,serum NT-proBNP,cTnI,and CK-MB levels in the model group were significantly increased.Myocardial tissue exhibited focal myocyte necrosis and dissolution,connective tissue hyperplasia,and an increased area of collagen fiber positivity.The cardiomyocyte apoptosis rate was elevated.Myocardial ATP levels were significantly decreased,while ADP and AMP levels were significantly increased.The activities of mitochondrial respiratory chain complexesⅠ-Ⅳwere markedly reduced.The Ca^(2+)fluorescence intensity in myocardial cells was significantly enhanced.Severe mitochondrial damage was observed,showing flocculent changes,increased numbers of damaged mitochondria,swelling,rupture,disordered arrangement,disrupted cristae,and partial vacuolation.The fluorescence intensity and protein expression levels of GPR35,p-AMPK,Bax,Caspase-3,Caspase-9,COL-Ⅰ,andα-SMA in myocardial tissue were significantly increased,whereas those of p-mTOR and Bcl-2 were decreased.Compared with the model group,AS-Ⅳsignificantly reduced serum NT-proBNP,cTnI,and CK-MB levels,markedly decreased the cardiomyocyte apoptosis rate,significantly increased ATP levels while reducing ADP and AMP content,enhanced the activities of mitochondrial respiratory chain complexesⅠ-Ⅳ,diminished Ca^(2+)fluorescence intensity,and alleviated mitochondrial damage,with mitochondria arranged more regularly.The fluorescence intensity and protein expression levels of GPR35,p-AMPK,Bax,Caspase-3,Caspase-9,COL-Ⅰ,andα-SMA were significantly decreased,while those of p-mTOR and Bcl-2 were increased.In conclusion,AS-Ⅳexerts an inhibitory effect on RIMF,and its mechanism may be associated with the regulation of the GPR35/AMPK/mTOR signaling axis to improve mitochondrial energy metabolism.