葛根通脉饮对兔动脉粥样硬化模型内质网应激信号转导通路相关分子表达的调控作用

The Regulation Effect of Kudzuvine Root Blood Circulation Promoting Drink in Endoplasmic Reticulum Stress Related Molecule Expression in Rabbit Atherosclerotic Lesions Model

目的探讨葛根通脉饮对兔动脉粥样硬化(AS)模型内质网应激(ERS)信号转导通路免疫球蛋白重链结合蛋白(BIP)mRNA及其蛋白表达的调控作用。方法选取健康日本大耳白兔32只,将其随机分为正常组、模型组、葛根通脉饮低剂量组(葛低组)、葛根通脉饮高剂量组(葛高组),每组8只。正常组饲喂基础饲料;模型组饲喂高胆固醇饲料(在基础饲料中加入1%胆固醇及0.02%蛋氨酸);葛低组、葛高组在饲喂高胆固醇饲料的同时,分别给予葛根通脉饮5 g·kg-1·d-1和10 g·kg-1·d-1。饲喂9周后,取兔胸主动脉上段,采用组织形态观察法观察胸主动脉病变情况,采用组织原位杂交及Western blotting杂交方法分别检测胸主动脉血管平滑肌细胞(VSMC)细胞质中BIP mRNA及其蛋白表达量。结果形态学观察显示,模型组血管病变典型,AS模型制备成功;葛低组、葛高组血管病变得到修复,其中葛高组血管修复效果明显。组织原位杂交及Western blotting杂交结果显示,4组兔胸主动脉VSMC BIP mRNA及其蛋白表达量间有差异(P<0.05);其中模型组、葛低组、葛高组BIP mRNA及其蛋白表达量高于正常组,葛低组、葛高组低于模型组,葛高组又低于葛低组(P<0.05)。结论葛根通脉饮可拮抗由同型半胱氨酸(Hcy)诱导的血管损伤,且高剂量的葛根通脉饮效果优于低剂量葛根通脉饮;葛根通脉饮抗AS作用机制与其对ERS通路BIP mRNA及其蛋白表达的调控有关。

Objective To investigate the effect of Kudzuvine Root blood circulation promoting drink（ KRBCPD） on regulating the expression of endoplasmic reticulum stress signaling pathway- related BIP mRNA and protein in rabbit atherosclerosis model.Methods A total of 32 healthy Japanese big- ear rabbits were selected as study subjects,the rabbits were divided into four groups： the normal control group,the model group,the low- dose KRBCPD group and the high- dose KRBCPD group,8 cases in each group.Cases in normal control group were fed with basic feed,cases in model group were fed with high cholesterol feed（ 1% cholesterol and 0.02% methionine were added in basic feed）,cases in low- dose KRBCPD were fed with high cholesterol feed and KRBCPD（ 5 g·kg^- 1·d^- 1）,cases in high- dose KRBCPD were fed with high cholesterol feed and KRBCPD（ 10 g·kg^- 1·d^- 1）.Nine weeks later,the superior segment of thoracic aorta was quickly obtained,the lesions in thoracic aorta were observed by form of organization observation method,the levels of BIP mRNA and protein which located in the cytoplasm of VSMC in thoracic aorta was detected by in situ hybridization and Western blotting hybridization,respectively.Results Morphological observation showed that vascular lesion in model group was typical,the AS model was successfully established.The vascular lesions in low- dose KRBCPD group and high- dose KRBCPD group got better,the repair effect in the high- dose KRBCPD group was better than that in the low- dose KRBCPD group.According to tissue in situ hybridization and Western blotting hybridization results,there were significant differences in levels of BIP mRNA and protein which located in the cytoplasm of VSMC in thoracic aorta among 4 groups（ P〈 0.05）; The levels of BIP mRNA and protein in model group,low- dose KRBCPD group and high- dose KRBCPD group were significantly higher than those in the normal control group,the levels of BIP mRNA and protein in low- dose KRBCPD group and high- dose KRBCPD group were significantly lower than those in model group,the levels of BIP mRNA and protein in high- dose KRBCPD group were significantly lower than those in low- dose KRBCPD group（ P〈 0.05）.Conclusion KRBCPD can repair Hcy- induced vascular injury,the effect of high- dose KRBCPD is better than that of low- dose KRBCPD,the anti- AS mechanism of KRBCPD is correlated with KRBCPD＇s regulation effect on ERS signaling pathway- related BIP mRNA and protein.