脾虚状态对高脂血症大鼠肝脏胆固醇代谢的影响及机制研究  

Study on the effects of spleen deficiency state on liver cholesterol metabolism in rat with hyperlipidemia and its mechanism

作　　者：朱美林[1] 贾连群[1] 杨关林[1] 李宁[1] 王俊岩[1] 杨祎[1] 杜莹[1] 宋囡[1] 冷雪[1] 

机构地区：[1]辽宁中医药大学省部共建中医脏象理论及应用教育部重点实验室,沈阳110032

出　　处：《中华中医药杂志》2015年第8期2712-2716,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金青年基金项目(No.81202834);国家重点基础研究发展计划(973计划)项目(No.201133CB531700);沈阳市科学技术计划项目(No.F-12-277-1-49)~~

摘　　要：目的:观察脾虚状态对高脂血症大鼠肝脏胆固醇代谢的影响并初步探讨其机制。方法:60只SPF级SD大鼠随机分为空白对照组、高脂血症组、脾虚高脂血症组。高脂血症组予高脂饲料喂饲,脾虚高脂血症组采用劳倦过度加饮食不节结合高脂饲料喂饲造模。间苯三酚法测定D-木糖排泄率,全自动生化分析仪检测血清胆固醇(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)、血清淀粉酶(AMY)水平,HE染色观察肝脏形态变化,油红O染色观察肝脏脂质沉积,电镜观察大鼠肝脏超微结构变化,实时定量反转录-聚合酶链反应(real-time RT-PCR)及Western Blot技术检测肝脏3-羟基-3-甲基戊二酸单酰辅酶A还原酶(HMGCR)、肝脂酶(HL)、胆固醇7α-羟化酶(CYP7A1)基因m RNA和蛋白表达。结果:与空白对照组比较,高脂血症组及脾虚高脂血症组血清TC、LDL-C水平显著升高(P<0.01),HDL-C、AMY水平显著降低,肝细胞形成大量脂质沉积,核周间隙增大,胞质内线粒体嵴减少,粗面内质网明显扩张,肝脏HMGCR、HL、CYP7A1 m RNA表达显著降低(P<0.01)。与高脂血症组相比,脾虚高脂血症大鼠血清TC、LDL-C水平显著升高(P<0.05,P<0.01),HDL-C、AMY及尿D-木糖排泄率显著降低,肝脏HL、CYP7A1 m RNA表达显著降低(P<0.05)。结论:脾虚运化功能失常引起血清胆固醇水平异常及肝脏脂质沉积,可能与调控肝脏HL和CYP7A1等基因表达有关。Objective： To observe the effects of spleen deficiency state on liver cholesterol metabolism in rat with hyperlipidemia and explore its mechanism. Methods： 60 SPF SD rats were randomly divided into blank control group, hyperlipidemia group and spleen deficiency combined hyperlipidemia group. Hyperlipidemia group were fed with high fat diet. Spleen deficiency combined hyperlipidemia group was established by using overstrain and improper diet with high fat diet. The phloroglucinol method was used to measure the rate of D-xylose excretion, and the levels of TC, TG, LDL-C, HDL-C and AMY in serum. HE staining and Oil Red O staining were used to observe the morphologic change and lipid accumulation in liver, respectively. Electron microscope was used to observe the ultrastructural changes in liver. Real-time PCR and western blot methods were used to detect the mRNA and protein of expression changes in HMGCR, HL and CYP7A1. Results： Compared with the blank group, the levels of TC and LDL-C in serum in the hyperlipidemia group and spleen deficiency combined hyperlipidemia group were significantly increased, while the level of HDL-C was decreased. In the hyperlipidemia group and spleen deficiency combined hyperlipidemia group, the result showed that the liver cells formed lots of lipid accumulation. Compared with the blank group, the perinuclear space of liver ceils increased, mitochondria crest in the cytoplasm increased, rough endoplasmic reticulum expanded and the mRNA expressions of HMGCR, HL and CYP7A1 in liver in the hyperlipidemia group and spleen deficiency combined hyperlipidemia group were decreased obviously. Compared with the hyperlipidemia group, the levels of TC and LDL-C in serum were increased, the HDL-C, AMY and D-xylose excretion were increased and the mRNA expressions of HL and CYP7A1 in liver decreased in spleen deficiency combined hyperlipidemia group. Conclusion： The digestion dysfunction induced by Spleen deficiency could cause disorder of cholesterol level in serum and lipid accumulation in

关 键 词：脾虚 胆固醇 高脂血症 肝脏 

分 类 号：R259[医药卫生—中西医结合]