摘要
目的:通过观察脾虚高脂血症(HLP)大鼠肝脏miRNA-122a及其与SREBP-2通路调控的关系,探讨脾虚对肝脏胆固醇代谢的影响及机制。方法:SD大鼠随机分为空白对照组、HLP组、脾虚HLP组。HLP组喂饲高脂饲料,脾虚HLP组同时予疲劳加饮食不节。造模结束后,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)、淀粉酶(AMY),HE及油红O染色观察肝脏脂质沉积,实时荧光定量PCR检测肝脏miRNA-122a及胆固醇调节元件结合蛋白2(SREBP-2)、羟甲基戊二酰辅酶A还原酶(HMGCR)、低密度脂蛋白受体(LDL-R)m RNA的表达,Western blotting检测肝脏上述蛋白。结果:与空白对照组比较,两组模型血清TC、TG、LDL-C水平显著升高,HDL-C、AMY水平显著降低(P<0.05,P<0.01),以后者改变显著(P<0.05,P<0.01);并且两组模型肝细胞均形成大量脂质沉积,肝脏miRNA-122a及其调控的基因和蛋白表达显著降低(P<0.05,P<0.01),以后者改变明显(P<0.05,P<0.01)。结论:脾虚加重血清胆固醇水平异常及肝脏脂质沉积,可能与miRNA-122a调控SREBP-2通路有关。
Objective： To investigate the influence and mechanism of spleen deficiency on cholesterol metabolism in liver through observing the expression of miRNA-122 a in the liver of hyperlipidemia（HLP） rats with syndrome of spleen deficiency and its regulation correlated with SREBP-2 signaling pathway. Methods： SD rats were randomly divided into blank control group, HLP group, and HLP with syndrome of spleen deficiency group. Rats in HLP group were fed with high fat diet, and rats in HLP with syndrome of spleen deficiency group received stimulation with fatigued combined with improper diet based on the feeding of high fat diet. After modeling, levels of serum TC, TG, LDL-C, HDL-C and AMY were detected. HE and oil red staining methods were used to observe the lipid accumulation of liver, PCR method was used to test the m RNA expressions of miRNA-122 a, SREBP-2, HMGCR and LDL-R, and Western blotting method was used to test the expression of the above proteins. Results： Campare with the blank control group, the levels TC, TG and LDL-C in serum increased significantly, while HDL-C and AMY levels in serum reduced significantly in rats of the HLP group and HLP with syndrome of spleen deficiency group（P0.05, P0.01）, the latter changes was significantly（P0.05, P0.01）. There were large amount of lipid accumulation in liver cells of rats in model group, while the expression of miRNA-122 a and the expression of genes and proteins which were regulated by miRNA-122 a in liver of rats in model groups decreased significantly（P0.05, P0.01）. Conclusion： The abnormal cholesterol levels in serum and the lipid accumulation in liver could be aggravated by spleen deficiency（P0.05, P0.01）, which might be related to SREBP-2 signal transduction pathway regulated by miRNA-122 a.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2016年第3期975-979,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
No.81300229)
国家重点基础研究发展计划(973计划)资助项目(No.2013CB531704)
辽宁省科学技术计划资助项目(No.2014225004)~~
