补阳还五汤抗动脉粥样硬化作用与紧密连接蛋白的关系

Relationship Between Anti-atherosclerosis Efffect and Tight Junction Protein by Buyang Huanwu Tang

摘要:

目的: 探讨补阳还五汤对同型半胱氨酸(homocysteine,Hcy)致动脉粥样硬化(AS) 的保护作用与紧密连接蛋白表达的关系. 方法: 将40只雄性载脂蛋白E基因敲除(ApoE^-/-)小鼠随机分为模型组、补阳还五汤低、中、高剂量组,Rho激酶抑制剂Y-27632组.以正常雄性C57BL/6J小鼠设正常组,每组8只动物.模型组和治疗组均采用0.1 g·kg^-1·d^-1的Hcy造模,造模同时,治疗组分别补阳还五汤(5,10,20 g·kg^-1·d^-1)组,Y-27632 50 mg·kg^-1组,ig.8周后采用Western-blot检测主动脉组织claudin蛋白的表达变化和反转录PCR(RT-PCR)检测连接蛋白包括咬合蛋白(occludin)、闭合蛋白(claudin)、带状闭合蛋白(zonula occludens, ZO)、连接黏附分子(junction adhesion molecule, JAM)的mRNA的表达变化. 结果: 与正常组对比,Western-blot检测模型组AS病变明显,模型组与正常组比claudin蛋白表达明显减少,RT-PCR检测模型组与空白组相比,claudin,JAM,Zo,Occludin的mRNA的表达明显降低,不同剂量的补阳还五汤改善AS病变的同时上调血管组织中Claudin,JAM,Zo,Occludin的表达水平. 结论: 补阳还五汤能显著的增加血管组织中Claudin,occludin,JAM,Zo表达量,说明补阳还五汤抗hcy所致的AS与增强血管组织中紧密连接蛋白有关.

Abstract:

Objective: To study the possible link between preventing effect of atherosclerosis (AS) and tight connection(TJ)Buyang Huanwu Tang(BYHWT). Method: Seventy-two male ApoE^-/-mice were randomly divided into five groups:model group, three dose groups of BYHWT(5,10,20 g·kg^-1·d^-1), Y-27632 group, and C57BL/6J mice was as a normal control group.AS model was established by intragastric administration of homocysteine(0.1 g·kg^-1·d^-1). Meanwhile,treatment groups were intragastrically administrated with BYHWT (5,10,20 g·kg^-1·d^-1),Y-27632 50 mg·kg^-1·d^-1. Eight weeks later, Western-blot and reverse transcription PCR(RT-PCR) were used to determine the expression of aortic claudin,junction adhesion molecule (JAM), zonula occludens (ZO). Result: Model group showed evident AS lesions.The expression of aortic claudin, JAM, ZO in the model group obviously decreased, compared with normal control group.Different doses of BYHWT could prevent atherosclerosis and up-regulate the claudin, JAM, ZO expressions. Conclusion: BYHWT could obviously increase the claudin, occludin,JAM, ZO expressions. BYHWT could prevent atherosclerosis induced by homocysteine via increasing the tight junction proteins expression.