基于PCR array技术探讨脾虚痰浊证大鼠肝脏脂蛋白和胆固醇代谢信号通路相关基因mRNA表达的变化  

Expression Changes of mRNA on Lipoprotein and Cholesterol Metabolism Signaling Pathways in Liver of Spleen Deficiency and Phlegm Turbidity Syndrome Rats by PCR Array Technology

作　　者：王英[1] 贾连群[1] 宋囡[1] 曹媛[1] 曹慧敏[1] 刘晶晶[1] 杨关林[1] 

机构地区：[1]辽宁中医药大学中医脏象理论及应用教育部重点实验室,辽宁沈阳110847

出　　处：《辽宁中医杂志》2016年第1期145-150,I0001,共7页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金青年基金项目(81202834)

摘　　要：目的:利用PCR array技术观测脾虚痰浊证大鼠肝脏脂蛋白和胆固醇代谢信号通路相关基因mRNA表达。方法:SPF级健康雄性SD大鼠20只,随机分为空白对照组、脾虚痰浊证组。脾虚痰浊证组采用劳倦过度加饮食不节结合高脂饲料喂饲造模。使用间苯三酚法测定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,提取各组大鼠肝脏总RNA,应用脂蛋白和胆固醇代谢信号通路PCR芯片检测脾虚痰浊证大鼠肝脏脂蛋白和胆固醇代谢信号通路相关基因mRNA表达。结果:与空白对照组相比,脾虚痰浊证大鼠血清HDL-C水平显著降低,TC和LDL-C水平显著升高,肝细胞形成大量脂质沉积。PCR array技术分析发现:与空白对照组相比,脾虚痰浊证大鼠上调≥2倍的基因8个,占10%,如Scarf1、Apol2、Abcg1、Apo A1、Acca2、RGD1564999、Il4;下调≥2倍的基因55个,占65%,如Cxcl16、Ldlr、Vldlr、Lrpap1、Ankra2、Apoa4、Cdh13、Colec12等,上述上调或者下调的基因,涉及脂蛋白信号通路的LDL受体、LDL受体相关蛋白、LDL相关蛋白及HDL相关基因;胆固醇转运途径的胆固醇转运、胆固醇流出以及胆固醇逆向转运相关基因;以及胆固醇代谢途径的胆固醇吸收、分解代谢、胆固醇的平衡及生物合成等相关基因。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,可能与调控肝脏的脂蛋白信号通路、胆固醇运输途径以及胆固醇代谢途径基因表达改变有关。Objective： To observe mRNA expression of genes that related to lipoprotein and cholesterol metabolism signaling pathways in spleen deficiency and phlegm turbidity syndrome rats by PCR array technology. Methods： Twenty SD rats were randomly divided into control group,spleen deficiency and phlegm turbidity syndrome group. Spleen deficiency and phlegm turbidity syndrome group used overstrain plus improper diet combined with high fat diet for modeling. D- xylose excretion rates were detected by phloroglucinol method and the contents of TG,TC,HDL- C,LDL- C and AMY in serum were detected by automatic biochemical analyzer. HE staining was used to observe the liver injury. Total RNA was extracted from livers in each group. PCR array was used to test mRNA expressions of genes that related to lipoprotein and cholesterol metabolism signaling pathways in livers. Results： Compared with control group,the levels of TC and LDL- C increased while the levels of HDL- C significantly decreased in spleen deficiency and phlegm turbidity syndrome group. HE staining indicated that obvious lipid deposition was formed in the liver cells. PCR array showed that the expression levels of 8（ 10%） genes（ e. g. Scarf1,Apol2,Abcg1,Apo A1,Acca2,RGD1564999,Il4） increased by double or more compared to those of the control group and the expression levels of 55（ 65%） genes（ e. g. Cxcl16,Ldlr,Vldlr,Lrpap1,Ankra2,Apoa4,Cdh13,Colec12,etc.） decreased by double or more compared to those of the control group. The above up and down- regulated genes involving Liporotein signaling including lipoprotein LDL receptors,LDL receptor related protein,protein involved in LDL and HDL related gene,cholesterol transport pathway related genes including cholesterol transport,cholesterol efflux and reverse cholesterol transport- relate,cholesterol metabolism pathways related genes including cholesterol absorption,catabolism,cholesterol balance and biosynthesis and other related genes. Conclusion： Abnormal serum cholesterol levels and liver lipid dep

关 键 词：脾虚 痰浊 脂蛋白 胆固醇 PCR ARRAY 

分 类 号：R332[医药卫生—人体生理学]