摘要
目的观察麝香酮调控脂肪酸合成酶(Fas)/脂肪酸合成酶配体(FasL)信号通路对糖尿病心肌病(DCM)大鼠心肌细胞凋亡的影响,探讨陈树泉“辛温通络”理论治疗DCM的作用机制。方法将SD大鼠分为空白组、DCM组、麝香酮低剂量组(0.68 mg/kg麝香酮)、麝香酮高剂量组(2.72 mg/kg麝香酮)、二甲双胍组(140 mg/kg二甲双胍)、麝香酮高剂量+重组人FasL蛋白(rh-FasL)组(2.72 mg/kg麝香酮+0.017 mg/kg rh-FasL),每组12只。除空白组外,其他组均采用高脂饲料喂养联合腹腔注射链脲佐菌素方式构建DCM大鼠模型,建模成功后,给药1次/d,持续6周。末次处理结束后,检测各组大鼠空腹血糖、左室射血分数(LVEF)、左室短轴缩短率(LVFS);采用苏木素-伊红(HE)染色法、Masson染色法分别检测心肌组织病理损伤及纤维化程度;采用分光光度法检测心肌组织中超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;采用TUNEL染色法观察心肌细胞凋亡情况;采用蛋白质印迹法(Western blot)检测心肌组织中裂解的天冬氨酸特异性半胱氨酸蛋白酶-3(Cleaved-caspase-3)、Bcl-2相关X蛋白(Bax)、Fas、FasL蛋白表达。结果与空白组比较,DCM组大鼠心肌组织病理损伤严重,空腹血糖、MDA含量、心肌胶原面积分数、心肌细胞凋亡率和Cleaved-caspase-3、Bax、Fas、FasL蛋白表达升高,LVEF、LVFS、SOD活性降低,差异有统计学意义(P<0.05);与DCM组比较,麝香酮低剂量组、麝香酮高剂量组、二甲双胍组大鼠心肌组织病理损伤减轻,空腹血糖、MDA含量、心肌胶原面积分数、心肌细胞凋亡率和Cleaved-caspase-3、Bax、Fas、FasL蛋白表达降低,LVEF、LVFS、SOD活性升高,差异有统计学意义(P<0.05);与麝香酮低剂量组比较,麝香酮高剂量组、二甲双胍组大鼠心肌组织病理损伤减轻,空腹血糖、MDA含量、心肌胶原面积分数、心肌细胞凋亡率和Cleaved-caspase-3、Bax、Fas、FasL蛋白表达降低,LVEF、LVFS、SOD活性升高,差异有统计学意义(P<0.05);与麝香酮高剂量组比较,麝香酮高剂量+rh-FasL组大鼠心肌组织病理损伤加剧,空腹血糖、MDA含量、心肌胶原面积分数、心肌细胞凋亡率和Cleaved-caspase-3、Bax、Fas、FasL蛋白表达升高,LVEF、LVFS、SOD活性降低,差异有统计学意义(P<0.05)。结论麝香酮可能通过抑制Fas/FasL通路抑制DCM大鼠氧化应激及心肌细胞凋亡。
Objective To observe the effect of muscone in regulating fatty acid synthase(Fas)/fatty acid synthase ligand(FasL)signaling pathway on cardiomyocyte apoptosis in rats with diabetic cardiomyopathy(DCM),and explore the mechanism of Chen Shuquan′s theory of"warming and clearing collaterals"in treating DCM.Methods SD rats were divided into blank group,DCM group,low-dose muscone group(0.68 mg/kg muscone),high-dose muscone group(2.72 mg/kg muscone),metformin group(140 mg/kg metformin),and high-dose muscone+recombinant human FasL protein(rh-FasL)group(2.72 mg/kg muscone+0.017 mg/kg rh-FasL),with 12 rats in each group.Except for the blank group,rats in all other groups were fed with high-fat feed combined with intraperitoneal injection of streptozotocin to construct a DCM rat model.After successful modeling,the rats were administered once a day for 6 weeks.After the last treatment,the changes in fasting blood glucose,left ventricular ejection fraction(LVEF),and left ventricular short axis shortening rate(LVFS)in rats were detected;Hematoxylin-eosin(HE)staining and Masson staining were applied to detect pathological damage and fibrosis degree of myocardial tissue,respectively.Spectrophotometry was applied to detect the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in myocardial tissue;TUNEL staining was applied to observe myocardial cell apoptosis;and Western blot was applied to detect the expression of cleaved aspartate specific cysteine protease-3(cleaved-caspase-3),Bcl-2 associated X protein(Bax),Fas,and FasL proteins in myocardial tissue.Results Compared with the blank group,the pathological damage to the myocardial tissue of rats in the DCM group was severe,the fasting blood glucose,MDA content,myocardial collagen area fraction and apoptosis rate of myocardial cell,and expression of cleaved-caspase-3,Bax,Fas,and FasL proteins increased,while the LVEF,LVFS,and activity of SOD decreased(P<0.05);compared with DCM group,pathological damage of myocardial tissue was reduced in the low-dose muscone group,the high-dose muscone group and the metformin group,and fasting blood glucose,MDA content,myocardial collagen area fraction,myocardial apoptosis rate and cleaved-caspase-3,Bax,Fas and FasL protein expression were decreased,and the activities of LVEF,LVFS and SOD were increased(P<0.05).Compared with low-dose muscone group,pathological damage of myocardial tissue was relieved in the high-dose muscone group and the metformin group,fasting blood glucose,MDA content,myocardial collagen area fraction,myocardial apoptosis rate,cleaved-caspase-3,Bax,Fas and FasL protein expression were decreased,and LVEF,LVFS and SOD activities were increased(P<0.05).Compared with the high-dose muscone group,the pathological damage to the myocardial tissue of rats in the high-dose muscone+rh-FasL group was intensified,the fasting blood glucose,MDA content,and expression of cleaved-caspase-3,Bax,Fas,and FasL proteins increased,the LVEF,LVFS,and activity of SOD decreased(P<0.05).Conclusion Muscone may inhibit oxidative stress and cardiomyocyte apoptosis in DCM rats by inhibiting the Fas/FasL pathway.
作者
魏芹
赵晓鹏
于华林
燕彩霞
WEI Qin;ZHAO Xiaopeng;YU Hualin;YAN Caixia(Department of Endocrinology,Jinan Integrated Traditional Chinese and Western Medicine Hospital,Jinan,Shandong,271100;the Second Department of Orthopedics,Jinan Integrated Traditional Chinese and Western Medicine Hospital,Jinan,Shandong,271100;Hepatology Department,Jinan Integrated Traditional Chinese and Western Medicine Hospital,Jinan,Shandong,271100;Oncology Department,Jinan Integrated Traditional Chinese and Western Medicine Hospital,Jinan,Shandong,271100)
出处
《实用临床医药杂志》
CAS
2023年第23期79-85,共7页
Journal of Clinical Medicine in Practice
基金
山东省中医药科技项目(鲁卫函〔2021〕46号)。
