基于氧化应激途径探讨人参益脉方干预血管老化作用机制  

Exploration on the Mechanism of Renshen Yimai Prescription in Preventing Vascular Aging Based on Oxidative Stress Pathway

作　　者：石丹丹 宁梓淇 郭晓晨 张瑶瑶 张元春[1] 张海燕 刘美霞[1] 刘剑刚[1] SHI Dandan;NING Ziqi;GUO Xiaochen;ZHANG Yaoyao;ZHANG Yuanchun;ZHANG Haiyan;LIU Meixia;LIU Jiangang(Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Graduate School of China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]中国中医科学院西苑医院,北京100091 [2]中国中医科学院研究生院,北京100700

出　　处：《中国中医药信息杂志》2024年第7期63-68,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074509);中国中医科学院科技创新工程项目(CI2021A01402)。

摘　　要：目的观察人参益脉方对ApoE^(-/-)小鼠氧化应激及血管老化的影响,探讨其干预血管老化的作用机制。方法将40只ApoE^(-/-)小鼠随机分为模型组、西药组(瑞舒伐他汀,2.6 mg/kg)及中药低、高剂量组(人参益脉方,4.29、8.58 g/kg),每组10只,另取10只C57BL/6J小鼠作为正常组。采用西式饮食饲料喂养ApoE^(-/-)小鼠建立血管老化模型,同时各给药组分别予相应药物灌胃,正常组和模型组灌胃等体积纯净水,连续12周。HE染色及Masson染色观察小鼠主动脉组织形态变化,ELISA检测血清氧化型低密度脂蛋白(ox-LDL)含量,比色法测定血清活性氧(ROS)、谷胱甘肽(GSH)、谷胱甘肽还原酶(GPX)、烟酰胺腺嘌呤二核苷酸(NAD^(+))含量,Western blot检测主动脉组织沉默信息调节因子1(SIRT1)、p21、p53、NADPH氧化酶4(NOX4)蛋白表达。结果与正常组比较,模型组小鼠主动脉可见脂质沉积,内膜及中膜明显增厚,弹性纤维减少,胶原纤维增多;血清ox-LDL、ROS含量显著升高(P<0.01),GSH、GPX、NAD^(+)含量显著降低(P<0.01);主动脉组织SIRT1蛋白表达显著降低(P<0.05),p21、p53蛋白表达显著升高(P<0.01,P<0.05)。与模型组比较,各给药组主动脉内膜可见少量脂质沉积,各层膜结构较清晰,胶原纤维减少;各给药组血清ox-LDL、ROS含量显著降低(P<0.01),GSH含量显著升高(P<0.05,P<0.01),中药低剂量组NAD^(+)含量显著升高(P<0.05);中药高剂量组主动脉组织p21、NOX4蛋白表达显著升高(P<0.05,P<0.01)。与西药组比较,中药高剂量组ROS含量显著降低(P<0.01),p53蛋白表达显著降低(P<0.05)。与中药低剂量组比较,中药高剂量组p21蛋白表达显著降低(P<0.01),NOX4蛋白表达显著升高(P<0.01)。结论人参益脉方可能通过调节氧化应激水平和相关蛋白表达减轻血管内皮损伤,从而发挥改善血管老化作用。Objective To observe the effects of Renshen Yimai Prescription on oxidative stress and vascular aging in ApoE^(-/-)mice;To explore its mechanism of intervention in vascular aging.Methods Forty ApoE^(-/-)mice were divided into model group,Western medicine group(rosuvastatin,2.6 mg/kg),TCM low-and high-dosage group(Renshen Yimai Prescription,4.29,8.58 g/kg),with 10 mice in each group.Another 10 C57BL/6J mice were set as normal group.A vascular aging model was established by ApoE^(-/-)mice fed with a Western diet.Each medication group was given corresponding drugs by gavage for 12 consecutive weeks,the normal group and model group were given equivalent volume of pure water.HE staining and Masson staining were used to observe the morphological changes of aortic tissue,and ox-LDL content in serum was detected by ELISA,the contents of ROS,GSH,GPX and NAD^(+)in serum were detected by colorimetric method,the expressions of SIRT1,p53,p21 and NOX4 protein in aortic tissue were detected by Western blot.Results Compared with the normal group,the model group mice showed significant fat deposition in the aorta,thickening of the intima and media,a significant decrease in elastic fibers,and an increase in collagen fibers;the serum contents of ox-LDL and ROS significantly increased(P<0.01),while the contents of GSH,GPX and NAD^(+)significantly decreased(P<0.01);the expression of SIRT1 protein in the aortic tissue significantly decreased(P<0.05),the expressions of p21 and p53 protein significantly increased(P<0.01,P<0.05).Compared with the model group,a small amount of lipid deposition was observed in the intima of aorta in each medication group,with clearer membrane structures in each layer and reduced collagen fiber;the serum contents of ox-LDL and ROS in each medication group were significantly decreased(P<0.01),while the GSH content significantly increased(P<0.05,P<0.01),the NAD^(+)content in TCM low-dosage group significantly increased(P<0.05);the expressions of p21 and NOX4 protein in aortic tissue of the TCM high-dosage gro

关 键 词：人参益脉方 血管老化 APOE-/-小鼠 氧化应激 

分 类 号：R285.5[医药卫生—中药学]