摘要
目的基于转化生长因子-β(transforming growth factor-β,TGF-β)/Smad信号通路探究黄芪葛根汤与远志汤合方抑制糖尿病动脉粥样硬化大鼠血管内皮细胞间质化的作用机制。方法将70只SPF级SD雄性大鼠随机分为空白对照组、模型组、阳性对照组(糖脉康颗粒)、黄芪葛根汤与远志汤合方1组(简称“合方1组”)、黄芪葛根汤与远志汤合方2组(简称“合方2组”)。空白对照组喂食普通饲料,第29天给予大鼠一次性腹腔注射柠檬酸缓冲液;其他组喂食高脂饲料,第29天给予大鼠一次性腹腔注射链脲佐菌素诱导2型糖尿病动脉粥样硬化模型,连续灌胃49 d。苏木精—伊红染色法观察主动脉的组织形态。检测各组大鼠血糖和血清总胆固醇(total cholesterol,TC)、三酰甘油(triglyceride,TG)、糖化血红蛋白(glycosylated hemoglobin A1c,HbA1c)、高密度脂蛋白胆固醇(high-density lipoprotein-cholesterol,HDL-C)、低密度脂蛋白胆固醇(low-density lipoprotein-cholesterol,LDL-C)水平;采用酶联免疫吸附试验检测血清波形蛋白(Vimentin)、Twist1、TGF-β、锌指转录因子1(Snail1)水平;采用Western blot法检测肝脏固醇调节元件结合蛋白-1c(sterol regulatory element-binding protein-1c,SREBP-1c)和主动脉Smad-2、Smad-3蛋白表达水平;采用qRT-PCR法检测葡萄糖转运蛋白4(glucose transporter-4,Glut-4)、Vimentin、腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)、SREBP-1c mRNA表达水平。结果与空白对照组比较,模型组大鼠血糖、HbA1c、TG、LDL-C、Vimentin、Twist1、TGF-β、Snail1、SREBP-1c、Smad-2、Smad-3表达水平显著升高(P<0.05),HDL-C、Glut-4、AMPK表达水平显著降低(P<0.05)。与模型组比较,合方1组、合方2组大鼠血糖、HbA1c、TG、TC、LDL-C、Vimentin、SREBP-1c、Twist1、TGF-β、Snail1、Smad-2、Smad-3水平显著降低(P<0.05),HDL-C、Glut-4、AMPK水平显著升高(P<0.05)。结论黄芪葛根汤与远志汤合方抑制主动脉内皮细胞间质化的作用与其参与主动脉TGF-β/Smad信号通路有关。
Objective To investigate the mechanism of action of Huangqi Gegen Decoction combined with Yuanzhi Decoction in inhibiting endothelial-mesenchymal transition in aortic endothelial cells of rats with diabetic atherosclerosis based on the transforming growth factor-β(TGF-β)/Smad signaling pathway.Methods A total of 70 specific pathogen-free male Sprague-Dawley rats were randomly divided into blank control group,model group,positive control group(Tangmaikang Granule),Huangqi Gegen Decoction+Yuanzhi Decoction group 1(combined prescription 1 group),and Huangqi Gegen Decoction+Yuanzhi Decoction group 2(combined prescription 2 group)according to blood glucose.The rats in the blank control group were fed with normal diet and were given a single intraperitoneal injection of citric acid buffer on day 29,and those in the other groups were fed with high-fat diet and were given a single intraperitoneal injection of streptozotocin on day 29 to induce a model of type 2 diabetic atherosclerosis,with intragastric administration for 49 consecutive days.HE staining was used to observe the histomorphology of the aorta;each group of rats were measured in terms of blood glucose and the serum levels of total cholesterol(TC),triglyceride(TG),glycosylated hemoglobin A1c(HbA1c),high-density lipoprotein-cholesterol(HDL-C),and low-density lipoprotein-cholesterol(LDL-C);ELISA was used to measure the serum levels of Vimentin,Twist1,TGF-β,and Snail1;Western blot was used to measure the protein expression level of sterol regulatory element-binding protein-1c(SREBP-1c)in the liver,as well as the protein expression levels of Smad-2 and Smad-3 in the aorta;qRT-PCR was used to measure the mRNA expression levels of glucose transporter-4(GLUT-4),Vimentin,AMP-activated protein kinase(AMPK),and SREBP-1c.Results Compared with the blank control group,the model group had significant increases in blood glucose and the expression levels of HbA1c,TG,LDL-C,Vimentin,Twist1,TGF-β,Snail1,SREBP-1c,Smad-2,and Smad-3(P<0.05)and significant reductions in the expression levels of HDL-C,GLUT-4,and AMPK(P<0.05).Compared with the model group,the combined prescription 1 group and the combined prescription 2 group had significant reductions in blood glucose,HbA1c,TG,TC,LDL-C,Vimentin,SREBP-1c,Twist1,TGF-β,Snail1,Smad-2,and Smad-3(P<0.05)and significant increases in the levels of HDL-C,GLUT-4,and AMPK(P<0.05).Conclusion The combination of Huangqi Gegen Decoction and Yuanzhi Decoction can inhibit endothelial-mesenchymal transition in aortic endothelial cells by participating in the aortic TGF-β/Smad signaling pathway.
作者
李茂宸
范颖
李洪洲
毕东雨
林庶茹
刘倩
LI Maochen;FAN Ying;LI Hongzhou;BI Dongyu;LIN Shuru;LIU Qian(School of Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Liaoning Shenyang 110847,China;Department of Traditional Chinese Medicine,Genertec Liaoyou Gem Flower Hospital,Liaoning Panjin 124010,China)
出处
《安徽中医药大学学报》
CAS
2023年第4期61-66,共6页
Journal of Anhui University of Chinese Medicine
基金
中国博士后科学基金资助项目(2021MD703843)
辽宁中医药大学重点实验室开放基金项目(zyzx2104)。
关键词
糖尿病动脉粥样硬化
内皮细胞间质化
黄芪葛根汤
远志汤
转化生长因子-β
Smad
信号通路
Diabetic atherosclerosis
Endothelial-mesenchymal transition
Huangqi Gegen Decoction
Yuanzhi Decoction
Transforming growth factor-β
Smad
Signaling pathway
