香砂六君子汤对脾虚高脂血症大鼠内质网应激PERK信号通路的影响

Effect of Xiangsha Liujunzi Decoction(香砂六君子汤)on PERK Signaling Pathway of Endoplasmic Reticulum Stress in Hyperlipidemia Rats with Spleen Deficiency

目的探讨香砂六君子汤对脾虚高脂血症大鼠内质网应激蛋白激酶R样内质网激酶(protein kinase R-like ER kinase,PERK)信号通路的影响。方法SD大鼠随机分为4组,即正常对照组、脾虚高脂组、香砂六君子汤组和辛伐他汀组,后3组采用饮食不节加力竭游泳的复合方法造模15 d后,正常组喂饲基础饲料,其余3组喂饲高脂饲料。12周后,分别给予相对剂量药物和生理盐水。4周后,检测各组大鼠血脂含量、D-木糖排泄率,油红O染色观察肝脏脂质沉积情况,实时荧光定量PCR及Western blot法检测大鼠肝脏葡萄糖调节蛋白78(glucose regulated protein 78 kD,GRP78)、PERK和活化转录因子4(the activating transcription factor 6,ATF4)基因及蛋白表达。结果与正常对照组比较,脾虚高脂组大鼠血清脂质异常,表现为总胆固醇(cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇酯(low density lipoprotein-cholesterol,LDL-C)明显升高,高密度脂蛋白胆固醇酯(high density lipoprotein-cholesterol,HDL-C)明显降低,尿D-木糖排泄率明显降低;肝脏脂质沉积显著,可见明显脂滴;肝脏GRP78、PERK和ATF4 mRNA水平及蛋白表达水平显著升高。与脾虚高脂组比较,香砂六君子汤组和辛伐他汀组大鼠血清TC、TG、LDL-C明显降低,HDL-C明显升高,尿D-木糖排泄率明显升高;其肝脏脂质沉积情况有明显改善,脂滴不同程度减轻;肝脏GRP78、PERK和ATF4 mRNA水平及蛋白表达水平明显降低。结论香砂六君子汤可能通过调控内网应激PERK信号通路来纠正脾虚脂质紊乱的状态。

Chinese Medicine Viscera State Theory and Applications,Liaoning University of Traditional Chinese Medicine;Objective To explore the effect of Xiangsha Liujunzi Decoction(香砂六君子汤)on protein kinase R-like ER kinase(PERK)signaling pathway of endoplasmic reticulum stress in hyperlipidemia rats with spleen deficiency.Methods SD rats were randomly divided into normal control group,hyperlipidemia group with spleen deficiency,Xiangsha Liujunzi Decoction group and simvastatin group.After 15 days of modeling by the compound method of diet and exhaustive swimming in the latter three groups,the normal group was fed with basal feed,and the other three groups were fed with high-fat feed.After 12 weeks,the Xiangsha Liujunzi Decoction group and the simvastatin group were given the relative doses of drugs,and the normal group and the hyperlipidemia group with spleen deficiency were given the corresponding volume of normal saline.After 4 weeks,the contents of total cholesterol(TC),triglyceride(TG),low density lipoprotein-cholesterol(LDL-C)and high density lipoprotein-cholesterol(HDL-C)were detected by automatic biochemical analyzer,the excretion rate of D-xylose was detected by the phloroglucinol method and the lipid deposition in the liver was observed by oil red O staining.The gene and protein expressions of glucose regulated protein 78 kD(GRP78),PERK and the activating transcription factor 4(ATF4)in rat liver were detected by real-time RT-PCR and Western blot.Results Compared with those of the normal control group,serum levels of TC,TG and LDL-C were significantly increased,the level of HDL-C was significantly decreased,and urinary D-xylose excretion rate was significantly decreased.GRP78,PERK and ATF4 mRNA and protein expression levels were significantly increased in the hyperlipidemia group with spleen deficiency.Compared with the hyperlipidemia group with spleen deficiency,the Xiangsha Liujunzi Decoction group and the simvastatin group significantly decreased serum levels of TC,TG and LDL-C,significantly increased HDL-C and significantly increased urinary D-xylose excretion rate.The lipid deposition was significantly improved,and lipid droplets were alleviated to varying degrees.The mRNA and protein expression levels of GRP78,PERK and ATF4 in the liver were significantly reduced.Conclusion Xiangsha Liujunzi Decoction may correct spleen deficiency and lipid disorder by regulating intranet stress PERK signaling pathway.