四逆散对代谢相关脂肪性肝病微循环障碍的影响及机制研究

Study on the effect and mechanism of Sini powder on the microcirculation disturbance of metabolic associated fatty liver disease

目的:探讨四逆散对代谢相关脂肪性肝病(MAFLD)微循环障碍的影响及作用机制。方法:将40只SD大鼠分为正常对照组10只和模型组30只,采用蛋氨酸和胆碱缺乏(MCD)饮食建立MAFLD模型。在造模成功之后,将模型组大鼠按照随机原则分为三组,分别为模型组、四逆散治疗组和水飞蓟素阳性药组,之后对正常对照组和模型组给予等体积0.9%氯化钠溶液灌胃,对四逆散治疗组和水飞蓟素阳性药组进行对应药物干预,连续灌胃4周后取材。分别对谷丙转氨酶(ALT)、谷草转氨酶(AST)等肝功能指标、超氧化物歧化酶(SOD)、丙二醛(MDA)等肝脏指标、内皮素1(ET-1)、一氧化氮(NO)等血清指标进行检测,HE染色和Masson染色观察大鼠肝脏组织病理学表现,超声造影观测肝脏微血管血液流速和状态,检测肠系膜微循环指标(微血流速度、微动静脉管径测量)和血液流变学指标(全血黏度、血浆黏度值变化、红细胞压积、红细胞流变学变化)。Westernblot法检测肝脏组织细胞间黏附分子-1(ICAM-1)、血管内皮细胞黏附分子-1(VCAM-1)和血管内皮生长因子(VEGF)蛋白表达。结果:与正常对照组相比,模型组大鼠血清中ALT、AST、ET-1、SOD水平显著提高,NO水平有所下降,MDA水平明显降低(P<0.05),造影剂到达肝动脉时间(HAAT)、造影剂到达肝静脉时间(HVAT)均有不同程度缩短(P<0.01),达峰时间(TP)提前,灌注参数峰值强度(PI)表现减弱,蛋白表达增加(均P<0.01)。与模型组比较,四逆散治疗组大鼠血清ALT、AST、ET-1水平有所下降(P<0.01),NO、SOD、MDA水平接近正常对照组(P<0.01),HAAT、HVAT有上升趋势(P<0.01),TP出现时间延长,PI表现增强(均P<0.01),蛋白表达水平降低。结论:四逆散可在一定程度上改善MAFLD的微循环障碍,其机制可能与抑制VCAM-1、ICAM-1和VEGF蛋白表达以降低血液黏稠度,从而改善血液淤滞情况有关。

Objective:To study the effect and mechanism of Sini powder on the microcirculation disturbance of metabolic associated fatty liver disease(MAFLD).Methods:A total of 40 SD rats were randomized divide into two groups,with 10 rats in the control group and 30 rats in the model group.The model group were induced MAFLD by methionine choline deficient(MCD)diet.After successfully establishing the models,the rats were randomly divided into Chinese herbal medicine group,positive drug group and model group of 8 each.The Chinese herbal medicine group was given Sini powder solution.The positive drug group was given the Silymarin powder suspension.The control group and the model group were given an equal volume of sodium chloride aqueous solution.Four weeks later,the rats were given anesthetic death for sampling.The pathological changes of liver in each group were observed by HE and Masson staining.The blood was collected to detect the liver function indicators[alanine aminotransferase(ALT),aspartate aminotransferase(AST)],liver biochemical indicators[superoxide dismutase(SOD),malondialdehyde(MDA)],serum index[endothelin-1(ET-1)],nitrous oxide(NO),contrast-enhanced ultrasound imaging was used to observe the blood flow rate and status of liver microvessels,mesenteric microcirculation indexes were detected(blood flow velocity arteriolar,blood flow velocity venules,arteriolar diameter,venule diameter)and hemorheology indexes(whole blood viscosity,plasmic viscosity,hematocrit,red blood cell rheology).Protein expression in liver tissues were detected by Western blot method[intercellular cell adhesion molecule-1(ICAM-1)、vascular cell adhesion molecule-1(VCAM-1)and vascular endothelial growth factor(VEGF)].Results:Compared with the control group,the serum levels of ALT,AST,ET-1 and SOD in the model group rats were significantly increased(P<0.01),NO and MDA levels were slightly decreased(P<0.05),HAAT and HVAT were shortened to varying degrees(P<0.01),TP appeared earlier,PI performance was weakened(P<0.01),and protein expression increased.Compared with the model group,the serum ALT,AST and ET-1 levels of the traditional Chinese medicine group rats decreased(P<0.01),NO and SOD with MDA levels were close to the control group,HAAT and HVAT showed an upward trend(P<0.01),TP appeared longer,PI expression increased(P<0.01),and protein expression levels decreased.Conclusion:Sini powder can improve microcirculation disorders of MAFLD to a certain extent,and its mechanism may be related to inhibiting the expression of VCAM-1,ICAM-1 and VEGF proteins,reducing blood viscosity,and thus clearing blood stasis.