健脾疏肝方对小鼠非酒精性脂肪性肝炎的干预作用

Intervention of Jianpi Shugan Decoction on non-alcoholic steatohepatitis in mice

目的:研究疏肝健脾方对非酒精性脂肪性肝炎(NASH)的干预作用及其机制。方法:40只雄性C57/BL6小鼠随机分为随机分为胆碱蛋氨酸充足(MCS)组、胆碱蛋氨酸缺乏(MCD)组、胆碱蛋氨酸缺含(MCD)组、健脾疏肝方(JP)组及复方蛋氨酸胆碱片(CMCB)组,每组10只。除MCS组外,采用MCD饮食诱导小鼠NASH模型。造模同时JP组小鼠予以疏肝健脾方中药煎剂灌胃给药2周(21.63g/kg),CMCB组予以CMCB溶液灌胃2周(274.6mg/kg),每日1次;MCS组和MCD组予以二甲基亚砜(DMSO)灌胃2周(1m L/10g),每日1次。2周后,处死动物,取血清和肝组织。观察肝组织病理学变化,检测肝功能、炎性因子、脂代谢、氧化应激及脂质过氧化相关指标,并进行相关因子的蛋白水平检测。结果:与MCD组比较,JP组肝组织病理变化明显改善,NAS病理评分显著降低(P=0.000);血清丙氨酸氨基转酶(ALT)活性、肿瘤坏死因子α(TNF-α)及白细胞介素6(IL-6)水平显著下降(P=0.000),肝组织甘油三酯(TG)(P=0.000)、丙二醛(MDA)水平(P=0.006)及核因子κB(NF-κB)表达(P=0.047)显著下降,肝组织肝性脂肪酸结合蛋白(L-FABP)表达显著升高(P=0.016)。CMCB组在降低血清ALT、TNF-α水平方面优于JP组(P<0.05),但在降低肝组织TG、血清TC及LDL方面效果不如JP组(P<0.05,P<0.01)。结论:健脾疏肝方通过多条途径对NASH起到正向干预作用。健脾疏肝方可抑制NF-κB的表达,抑制炎性因子的生成,调控炎性信号传导通路;并可通过增加L-FABP表达调控肝组织脂质转运及代谢,减少氧化应激反应,抑制脂质过氧化。

Objective: To investigate the regulation mechanism of Jianpi Shugan Decoction on murine non-alcoholic steatohepatitis. Methods: Forty mice were randomly divided into four groups. Experimental NASH was induced in mouse by the methionine-choline-deficient diet. JP group was treated with Jianpi Shugan Decoction solution of curcumin(21.63g/kg) via intragastric administration once a day for 2 weeks. CMCB group was treated with DMSO solution of compound methionine and choline bitartrate tablets(274.6mg/kg). MCS group and MCD group were treated with DMSO. After 2 weeks mice were killed to obtain blood samples and liver tissues. Hepatic pathological changes were observed; ALT, AST, TG, TC, HDL, LDL were assessed by biochemical measurements; TNF-α, IL-6, IL-4 was determined by radioimmunoassay method; NF-κB, L-LABP protein expression was estimated by Western blot. Results: Compared with MCD the pathological changes of liver tissue in group JP was obviously improved, the pathological score decreased significantly(P=0.000); the activity of serum ALT, TNF-α and IL-6 levels decreased significantly(P=0.000); liver tissue TG(P=0.000), MDA levels(P<0.006) and the expression of NF-kappa B(P=0.047) decreased significantly; the expression of L-FABP levels were significantly higher(P=0.016). CMCB group was better than JP group in reducing serum ALT and TNF-αlevel(P<0.05), but the effects to the liver tissue TG and the serum LDL were lower than that of JP group(P<0.05, P<0.01). Conclusion: Jianpi Shugan Decoction has positive effect on NASH through multiple ways. Jianpishugan decoction can inhibit the expression of NF-κB, suppress the generation of inflammatory factors, play a role in regulation of inflammatory signal transduction pathway; and by increasing the expression of L-FABP and the role in the regulation of lipid transport and metabolism of liver tissue expression; in addition Jianpishugan decoction can reduce oxidative stress, inhibition of lipid peroxidation.