香砂六君子汤通过调控miR-182/FOXO1对脾虚高脂血症大鼠肝脏脂质沉积的影响及可能的机制

Effects of Xiangsha Liujunzi Decoction on Liver Lipid Deposition in Rats with Spleen Deficiency and Hyperlipidemia by Regulating mi R-182/FOXO1 and Its Possible Mechanism

目的观察miR-182及叉头转录因子-1(FOXO1)介导的高密度脂蛋白(HDL)代谢在脾虚高脂血症大鼠肝脏中及给予香砂六君子汤后大鼠肝脏中的变化,探讨香砂六君子汤干预肝脏HDL代谢进而影响脂代谢的机制。方法SD大鼠28只随机分为四组:分别为正常组、高脂血症对照组、脾虚高脂血症组、香砂六君子汤组。从造模开始起饲喂14周,常规饲料饲喂正常组,高脂饲料饲喂剩余其他各组;饲喂开始前15天,脾虚高脂血症组、香砂六君子汤组建立常规脾虚模型(饮食不节加劳倦过度复合方法);饲喂10周后,开始进行灌胃给药:香砂六君子汤11.34 g/kg/天,其他各组生理盐水(同体积)。14周后进行取材检测:血脂水平常规件检测;病理学技术检测肝脏病理变化;ELISA法进行载脂蛋白A1(APOA1)、载脂蛋白B(APOB)检测;肝脏miR-182、FOXO1、B族Ⅰ型清道夫受体(SR-BI)、肝脂酶(HL)、固醇调节元件结合蛋白2(SREBP-2)mRNA含量变化通过实时荧光定量PCR法进行检测。结果高脂血症对照组及脾虚高脂血症组大鼠血脂及ApoA-Ⅰ、ApoB水平变化显著;总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、ApoB含量上升显著(P<0.01),高密度脂蛋白胆固醇(HDL-C)、ApoA-Ⅰ含量降低显著(P<0.01);肝脏出现油脂沉积及空泡;FOXO1 mRNA含量及蛋白表达增加;miR-182 mRNA含量显著下降(P<0.01),miR-182/FOXO1介导的HDL代谢下游基因HL、SR-BI、SREBP-2mRNA表达降低(P<0.01);脾虚引起以上指标变化程度加深(P<0.05,P<0.01);干预给药后,各指标得到不同缓解(P<0.01)。结论脾运化功能失常可能与miR-182/FOXO1介导HDL代谢途径异常有关,香砂六君子汤干预机制可能与上述途径相关。

Objective To observe the high-density lipoprotein(HDL)metabolism mediated by miR-182 and forkhead transcription factor-1(FOXO1)in the liver of spleen-deficiency hyperlipidemia rats and the liver of rats after Xiangsha Liujunzi Decoction was administered,and to explore the mechanism of Xiangsha Liujunzi Decoction interfering with liver HDL metabolism and then affecting lipid metabolism.Methods 28 SD rats were randomly divided into four groups:normal group,hyperlipidemia control group,spleen deficiency hyperlipidemia group,Xiangsha Liujunzi Decoction group.Feeding for 14 weeks from the start of model building,the normal group was fed with regular feed,and the remaining groups were fed with high-fat feed;15 days before the start of feeding,the spleen deficiency and hyperlipidemia group and the Xiangsha Liujunzi Decoction group established routine spleen deficiency Model(compound method of unregulated diet and excessive fatigue);after feeding for 10 weeks,start gavage:Xiangsha Liujunzi Decoction 11.34 g/(kg·d),other groups of physiological saline(same volume).After 14 weeks,the samples were collected and tested:routine blood lipid level detection;pathological techniques to detect liver pathological changes;ELISA method for apolipoprotein A1(APOA1)and apolipoprotein B(APOB)detection;liver miR-182,FOXO1,type B scavenger receptor(SR-BI),liver lipase(HL),sterol regulatory element binding protein 2(SREBP-2)mRNA content changes were quantified by real-time fluorescence PCR method for detection.Results The blood lipids,ApoA-Ⅰand ApoB levels of the hyperlipidemia control group and the spleen-deficiency hyperlipidemia group changed significantly:total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),ApoB content increased significantly(P<0.01),high-density lipoprotein cholesterol(HDL-C),ApoA-Ⅰcontent decreased significantly(P<0.01);lipid deposition and vacuoles appeared in the liver;FOXO1 mRNA content And protein expression increased;miR-182 mRNA content was significantly decreased(P<0.01),miR-182/FOXO1 mediated HDL metabolism downstream genes HL,SR-BI,SREBP-2mRNA expression decreased(P<0.01);spleen deficiency caused the deepened change of above index(P<0.05,P<0.01);after intervention administration,each index was relieved differently(P<0.01).Conclusion The dysfunction of spleen transport and chemistry may be related to the abnormal pathway of HDL metabolism mediated by miR-182/FOXO1,and the intervention mechanism of Xiangsha Liujunzi Decoction may be related to the above pathway.