补阳还五汤调控衣霉素诱导的ApoE^(-/-)小鼠内质网应激从而减少动脉粥样硬化易损斑块面积  

Buyang Huanwu Decoction Regulates Tunicamycin Induced Endoplasmic Reticulum Stress in ApoE^(-/-) Mice and Reduces the Area of Vulnerable Plaque in Atherosclerosis

作　　者：王海贝 容子玲 张路煜 白锦婷 刘玉晖[1] 游宇[2] WANG Haibei;RONG Ziling;ZHANG Luyu;BAI Jinting;LIU Yuhui;YOU Yu(Jiangxi University of Traditional Chinese Medicine,Nanchan 330004 Jiangxi,China;First Affiliated Hospital of Nanchang University,Nanchang 330006 Jiangxi,China)

机构地区：[1]江西中医药大学,江西南昌330004 [2]南昌大学第一附属医院,江西南昌330006

出　　处：《中药新药与临床药理》2022年第6期721-726,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81860714)。

摘　　要：目的基于IRE1-XBP1通路探究补阳还五汤(BYHWT)抑制由衣霉素诱导的载脂蛋白E基因敲除小鼠(ApoE^(-/-)小鼠)内质网应激从而提高动脉易损斑块稳定性的作用及机制。方法高脂饲料喂养ApoE^(-/-)小鼠12周制备动脉粥样硬化(AS)模型,在AS模型基础上腹腔注射衣霉素0.9 mg·kg^(-1)。造模后将小鼠分为模型组,4-PBA组(给药剂量为0.5 g·kg^(-1)),补阳还五汤低、中、高剂量组(给药剂量为5、10、20 g·kg^(-1)),每组10只,继续高脂喂养。以相同遗传背景非转基因小鼠(C57BL/6J小鼠)作为正常对照组(给蒸馏水),给予普通饲料喂养。8周后收集主动脉进行苏木素-伊红染色观察;TUNEL法检测主动脉内细胞凋亡情况;ELISA法检测血清白细胞介素1β(1L-1β)和白细胞介素6(IL-6)的水平;Western Blot法检测内质网应激标志性蛋白内质网分子伴侣糖调节蛋白(GRP78)、内质网跨膜激酶1(IRE1)、X盒结合蛋白(XBP1)及凋亡相关蛋白Bcl-2相关X蛋白(BAX)、B细胞淋巴瘤-2(Bcl-2)、C/EBP同源蛋白(CHOP)的表达量。结果与正常组比较,模型组斑块面积明显增加;与模型组比较,4-PBA组与补阳还五汤低、中、高剂量组血管内斑块均有减少,主动脉斑块内细胞凋亡率下降(P<0.01)。与正常组比较,模型组IL-6与IL-1β水平均升高(P<0.01);与模型组比较,补阳还五汤高剂量组及4-PBA组IL-1β水平均下降(P<0.05),补阳还五汤低、中、高剂量组及4-PBA组IL-6水平均下降(P<0.05)。与正常组比较,模型组内质网应激相关蛋白GRP78、IRE1、XBP1相对蛋白表达含量均有上升(P<0.05);与模型组比较,补阳还五汤低、中、高剂量组及4-PBA组内质网应激标志性蛋白GRP78、IRE1、XBP1相对蛋白表达量均有下降(P<0.05)。与正常组比较,模型组凋亡蛋白BAX与CHOP相对表达量均明显上升(P<0.05),Bcl-2相对表达量下降(P<0.05);与模型组比较,补阳还五汤中、高剂量组与4-PBA组BAX、CHOP相对表达量均有下降(P<0.05),BObjective To explore the effect and mechanism of Buyang Huanwu Decoction(BYHWT)on enhancing stability of artery vulnerable plaques via inhibiting endoplasmic reticulum stress induced by tunicamycin in apolipoptotein E-knockout mice(ApoE^(-/-)mice)based on IRE1-XBP1 pathway.Methods ApoE^(-/-)mice were fed high-fat diet for 12 weeks to make atherosclerosis(AS)model.Then the mice were intraperitoneously injected with 0.9 mg·kg^(-1)tunicamycin.After modeling,the mice were divided into model group,4-PBA group(0.5 g·kg^(-1)),low-,medium-and high-dose BYHWT group(5,10,20 g·kg^(-1)),10 mice in each group,which continued receiving high fat diet.Non-transgenic mice(C57BL/6J mice)with the same genetic background were used as normal control group,which were given distilled water and normal diet.Eight weeks later,the aorta was collected for hematoxylineosin staining,and TUNEL assay was used to detect the cell apoptosis in the aorta.Serum interleukin-1β(IL-1β)and interleukin-6(IL-6)levels were determined by ELISA.Endoplasmic reticulum stress marker molecular chaperone glycoregulatory protein(GRP78),endoplasmic reticulum transmembrane kinase 1(IRE1),X-box binding protein(XBP1),apoptosis related protein Bcl-2 associated X protein(BAX),B-cell lymPhoma-2(Bcl-2)and C/EBP homologous protein(CHOP)were detected by Western Blot.Results Compared with the normal group,the plaque area in the model group increased significantly.Compared with the model group,the intravascular plaque in 4-PBA group,low-,medium-and high-dose BYHWT groups decreased,and the apoptosis rate in the aortic plaque decreased(P<0.01).Compared with the normal group,IL-6 and IL-1βlevels in model group were increased(P<0.01),Compared with the model group,IL-1βin high-dose BYHWT group and 4-PBA group showed a downward trend(P<0.05),and IL-6 levels in low-,medium-,high-dose BYHWT groups and 4-PBA group also had a downward trend(P<0.05).Compared with normal group,the relative protein expression levels of GRP78,IRE1 and XBP1 in model group were increased(P<0.05).Compa

关 键 词：补阳还五汤 衣霉素 内质网应激 动脉粥样硬化 易损斑块 小鼠 

分 类 号：R285.5[医药卫生—中药学]