摘要
目的观察化瘀祛痰方对动脉粥样硬化(AS)家兔肝脏线粒体自噬相关蛋白的影响,探讨线粒体自噬对肝脏脂质沉积的调控作用。方法 SPF级健康雄性新西兰纯种家兔60只,随机分为正常组、模型组、化瘀祛痰组、辛伐他汀组,正常组喂饲基础饲料,模型组、化瘀祛痰组、辛伐他汀组喂饲高脂饲料造模。造模8周后,化瘀祛痰组、辛伐他汀组给予相应药物,正常组及模型组给予相应体积生理盐水。4周后,检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)含量,HE染色观察肝脏组织的形态变化,油红O染色观察脂滴蓄积,Western blotting检测肝脏组织线粒体自噬相关蛋白FUNDC1、P62及LC3的相对表达量。结果与正常组相比,模型组家兔血清TC、TG、LDL-C含量显著上升(P<0.01),HDL-C含量显著降低(P<0.05);与模型组相比,化瘀祛痰组及辛伐他汀组血清TC、TG、LDL-C含量显著下降(P<0.05),HDL-C含量显著上升(P<0.05)。HE染色结果,模型组家兔肝脏组织肿胀明显,脂肪空泡形成;化瘀祛痰组及辛伐他汀组脂肪空泡显著减少或消失,肝组织形态恢复或部分恢复正常。油红O染色结果,模型组肝细胞肿大变圆,胞浆疏松,胞浆内脂滴蓄积明显;化瘀祛痰组及辛伐他汀组脂肪变性程度降低,脂质空泡数量减少。Western blotting结果,与正常组相比,模型组家兔肝组织中线粒体自噬相关蛋白LC3、FUNDC1相对表达量显著升高(P<0.05),P62相对表达量显著降低(P<0.05),而化瘀祛痰组及辛伐他汀组与模型组相比线粒体自噬相关蛋白LC3、FUNDC1相对表达量显著降低(P<0.05),P62相对表达量显著升高(P<0.05)。结论化瘀祛痰方可调控线粒体自噬相关蛋白,减少AS家兔肝组织脂质沉积,减缓动脉粥样硬化的形成和发展。
Objective To observe the effect of stasis-removing phlegm-resolving formula（ Huayu QutanFang,HYQTF） on liver mitochondrial autophagy related proteins,and to investigate the effects of mitochondrial autophagy on liver lipid deposition in rats with atherosclerosis（ AS）. Methods Rabbits（ 60-SPF-grade Zelanian purebred males） were randomly divided into normal group,model group,HYQTF group and simvastatin group. Normal group was fed with normal diet; model group,HYQTF group and simvastatin group were fed with high-fat diet. 8 weeks after the modeling,HYQTF group and simvastatin group received corresponding medicinal intervention while normal group and model group were administered with the same volume of saline. After 4 weeks,the serum contents of total cholesterol（ TC）,triglyceride（ TG）,low-density lipoprotein-cholesterol（ LDL-C） and high-density lipoprotein-cholesterol（ HDL-C）were measured whereas the morphological changes of rabbit liver tissue were observed by using stained HE.Accumulation of lipid droplets was evaluated with oil red O staining,and the relative expression of autophagy relating protein FUNDC1,P62 and LC3 in rabbit liver tissue were detected with Western blotting method. Results Compared with normal group,TC,TG and LDL-C levels of serum increased significantly（ P〈0. 01） whereas HDL-C level decreased（ P〈0. 05） in the model group. Compared with the model group,serum TC,TG and LDL-C levels in the HYQTF group and simvastatin group decreased significantly（ P〈0. 05）,and the content of HDL-C increased significantly（ P〈0. 05）. HE staining results： The liver tissue of model group rabbits became apparently swollen and formed fatty vacuolization. Fatty vacuoles in HYQTF group and simvastatin group significantly decreased or disappeared. Normal morphology of liver tissue fully or partially recovered. Oil Red O staining results： In the model group,liver cell became swollen and round,with the loose cytoplasm and obvious accumulation of intracellular lipid droplets. The degree of steatosis decreased in HYQTF group and simvastatin group,with the number of lipid vacuoles decreased. Western blotting results： Compared with normal group,the relative expression of mitochondrial autophagy relating proteins LC3 and FUNDC1 in liver tissue of rabbits significantly increased in the model group（ P〈0. 05）,and the relative expression level of P62 decreased significantly（ P〈0. 05） in the model. Compared with the normal group, mitochondrial autophagy relating protein LC3, FUNDC1 expression in model group increased significantly（ P〈0. 05）,while mitochondrial autophagy relating protein LC3,FUNDC1 expression in HYQTF group and simvastatin group decreased significantly,and the relative expression level of P62 increased significantly（ P〈0. 05）. Conclusion Stasis-removing phlegmresolving formula may reduce lipid deposition in liver tissue of AS rabbits by regulating mitochondrial autophagy related proteins,thereby slowing the formation and development of atherosclerosis.
作者
陈丝
贾连群
宋囡
王健
陈宁
高云飞
谢思梦
Chen Si;Jia Lianqun;Song Nan;Wang Jian;Chen Ning;Gao Yunfei;Xie Simeng(Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications of Liaoning University of Traditional Chinese Medicine, Liaoning 110847, China;Translational medicine research center of Traditional Chinese medicine, Liaoning 110847, China)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2018年第4期310-315,共6页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家自然科学基金项目(No.81774022)
辽宁省百千万人才工程资助项目(No.2017-58)
辽宁省高等学校优秀人才支持计划项目(No.LR2015041)
沈阳市科技计划项目(No.17-139-8-00)~~
关键词
化瘀祛痰方
线粒体自噬
脂质沉积
动脉粥样硬化
家兔
Huayu Qutan Formula
mitochondrial autophagy
lipid deposition
atherosclerosis
rabbits
