化瘀祛痰方对脾虚型高脂血症大鼠肝脏SREBP-2信号通路的干预作用  

Intervention of Huayu Qutan Recipe on Liver SREBP-2 Signal Pathway of Hyperlipidemia Rats of Pi Deficiency Syndrome

作　　者：冷雪[1] 贾连群[1] 杨关林[1] 王俊岩[1] 朱美林[1] 王莹[1] 徐荧[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：《中国中西医结合杂志》2015年第3期320-326,共7页Chinese Journal of Integrated Traditional and Western Medicine

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

摘　　要：目的探讨化瘀祛痰方对脾虚型高脂血症大鼠肝脏固醇反应元件结合蛋白-2(sterol regulatory element binding protein-2,SREBP-2)信号通路的干预作用。方法 100只SPF级SD大鼠随机分为空白对照组、高脂血症组、高脂血症治疗组、脾虚型高脂血症组、脾虚型高脂血症治疗组,每组20只。空白对照组给予普通大鼠颗粒饲料喂养;高脂血症组和高脂血症治疗组给予高脂饲料;脾虚高脂血症和脾虚高脂血症治疗组采用劳倦过度和饮食不节进行造模,单日除饲喂高脂饲料外,予精炼猪油灌胃,每次3 m L,每日2次,双日喂食甘蓝不限量,造模时间30天。高脂血症治疗组和脾虚高脂血症治疗组灌胃化瘀祛痰方煎剂(20 m L/kg),1次/天,连续30天。全自动生化分析仪检测血清TC、TG、LDL-C、HDL-C、血清淀粉酶(AMY)水平,间苯三酚法测定D-木糖排泄率,HE染色观察肝脏形态变化,油红O染色观察肝脏脂质沉积,实时定量反转录-聚合酶链反应(Real time PCR)及Western blotting技术检测HMGCR、CYP7A1、LDL-R、SREBP-2 mRNA和蛋白表达。结果与空白对照组比较,高脂血症组及脾虚型高脂血症组血清TC(mmol/L,1.84±0.19,2.23±0.43)、LDL-C水平(mmol/L,0.99±0.24;1.13±0.56)显著升高(P<0.05),HDL-C水平(mmol/L,0.41±0.66;0.41±0.11)及AMY活性(mmol/L,351±45;153±30)显著降低(P<0.05),尿D-木糖排泄率(ng/mL,26.9±2.1;15.0±1.7)显著降低(P<0.05),肝细胞形成脂质沉积,胞浆内多出现脂肪空泡,肝脏CYP7A1、HMGCR、LDL-R、SREBP-2 mRNA及蛋白表达显著降低(P<0.01)。与高脂血症组比较,脾虚高脂血症大鼠血清TC、LDL-C水平显著升高(P<0.05),AMY及尿D-木糖排泄率显著降低(P<0.01),肝脏形成大量脂质沉积,肝细胞萎缩明显,肝脏CYP7A1、LDL-R、SREBP-2 mRNA及蛋白表达降低(P<0.01,P<0.05);高脂血症治疗组TC、LDL-C含量降低(P<0.05),HDL-C含量、AMY活性及D-木糖排泄率升高(P<0.05),CYP7A1、LDL-R、SREBP-2 mRNA及蛋白表达升高(P<0.01)。�Objective To explore the intervention of Huayu Qutan Recipe （HQR） on liver SREBP-2 signal pathway of hyperlipidemia rats of Pi deficiency syndrome （PDS）. Methods Totally 100 SPF grade SD rats were randomly divided into the blank control group, the hyperlipidemia group, the hy- perlipidemia treatment group, the PDS hyperlipidemia group, and the PDS hyperlipidemia treatment group, 20 in each group. Common granular forage was fed to rats in the blank control group. High fat for- age was fed to rats in the hyperlipidemia group and the hyperlipidemia treatment group. Rats in the PDS hyperlipidemia group and the PDS hyperlipidemia treatment group were treated with excessive labor and improper diet for modeling. They were administered refined lard by gastrogavage （3 mL each time, twice per day） and fed with high fat forage on the odd days, and fed with wild cabbage freely on even days. The modeling lasted for 30 days. Rats in the hyperlipidemia treatment group and PDS hyperlipidemia treatment group were administered with Huayu Qutan Recipe （20 mL/kg） by gastrogavage, once a day, for 30 suc- cessive days. Levels of serum cholesterol （TC）, triglyceride （TG）, low density lipoprotein cholesterol （LDL-C）, high density lipoprotein cholesterol （HDL-C）, and serum amylase （AMY） were detected by automatic biochemical analyzer. D-xylose excretion rate was determined using phloroglucinol method. Morphological changes of liver and the lipid deposition in liver were observed using HE stain and oil red O stain respectively, mRNA and protein expression levels of 3-hydroxy-3-methylglutaryl coenzyme A reduc- tase （HMGCR）, cholesterol 7α-hydroxylase 1 （CYP7A1）, LDL-R, and sterol regulatory element binding protein-2 （SREBP-2） were detected using real time RT-PCR and Western blotting. Results Compared with the blank control group, serum levels of TC （1.84 ±0.19 mmol/L,2.23 ±0.43 mmol/L） and LDL-C （0.99±0.24 mmol/L, 1.13 ±0.56 mmol/L） were higher in the hyperlipidemia group and the

关 键 词：脾虚 高脂血症 固醇反应原件结合蛋白2 化瘀祛痰方 胆固醇 

分 类 号：R285.5[医药卫生—中药学]