灵甲方对血瘀型肝纤维化大鼠骨桥蛋白的干预研究

To Study the Effect of Intervention of LingJiaFang on Expression of Osteopontin in Blood Stasis Type of Hepatic Fibrosis

目的:了解灵甲方对血瘀型肝纤维化动物模型的防治肝纤维化作用,从OPN、PAI-1的蛋白表达层面探索其防治肝纤维化的机理。方法:将Wistar大鼠共70只,随机分为4组:正常对照组、灵甲方组、复方鳖甲软肝片组、血瘀组。除正常对照组外,其余3组均采用腹腔注射DMN同时尾静脉注射小牛血清白蛋白和去甲肾上腺素的方法,复制血瘀型肝纤维化大鼠模型共4周。4周后灵甲方组以灵甲方灌胃,每天1次,共治疗8周。复方鳖甲软肝片对照组4周后以复方鳖甲软肝片灌胃,每天1次,共治疗8周。8周结束后,取血检测ALT、AST、Alb、Glb;取肝组织,用Western blot检测OPN与PAI-1的蛋白表达。结果:与血瘀组比,灵甲方及复方鳖甲软肝片治疗后大鼠的ALT明显降低(P<0.05),且灵甲方组的AST较血瘀组降低明显(P<0.01);与血瘀组比,灵甲方治疗后大鼠的体重明显增加(P<0.01),复方鳖甲软肝片治疗后大鼠的体重亦有增加(P<0.05);与血瘀组比,灵甲方治疗后肝体比比值显著增高(P<0.01),脾体比比值降低(P<0.05),且灵甲方组较复方鳖甲软肝片治疗组在治疗后肝体比比值增加更明显(P<0.05)。灵甲方治疗后OPN及PAI-1的蛋白表达水平均较血瘀组显著降低(P<0.01),灵甲方较复方鳖甲软肝片对OPN表达的抑制作用更强,差别有统计学意义(P<0.05)。结论:灵甲方可能通过抑制OPN的表达间接降低PAI-1的蛋白表达水平,所以表现为对PAI-1蛋白表达水平的降低幅度较OPN大,从而使PAI-1对细胞外基质降解的抑制作用减弱,增加细胞外基质的降解,发挥抗纤维化作用。

Objectives：To investigate the effect of intervention of LingJiaFang in hepatic fibrosis through the liver of a rat model of blood stasis type hepatic fibrosis,and its influence on protein expression of OPN,PAI-1.Methods：The rat model of blood stasis type hepatic fibrosis was induced by DMN BSA and NE.Fifty male Wistar rats were randomly divided into 4 groups：normal group,XueYu,LingJiaFang and FuFang BieJia RuanGan Pian.After 8 weeks consecutive treatment,all rats were sacrificed and blood and liver tissue specimens were obtained.Liver functions（ALT AST）were determined.Histochemical staining of formalin-fixed liver sections with HE.Collagen deposition in liver was studied by Sirius red staining.Protein expression of osteopontin（OPN）and PAI-1 in liver tissue were tested with Western blot and all the dates were analyzed with SPSS11.0 statistical software.Results：Compared with the XueYu group,the serum levels of ALT in the treated groups were significantly decreased（P0.05）,and the serum levels of AST in LinJiaFang group were obviously decreased（P0.01）.The growing weight was also higher in LinJiaFang group than that in FuFangBieJiaRuanGanPian group（P0.01）.Compared with XueYu group,the expression of OPN and PAI-1 was greatly decreased by LingJiaFang（P0.01）,while the expression of OPN and PAI-1 in FuFangBieJiaRuanGanPian group was also decreased（P0.05）.But the inhibition effect of LinJiaFang on the expression of OPN stronger than FuFangBieJiaRuanGanPian（P0.05）.Conclusion：LingJiaFang can decrease the expression of OPN and PAI-1,maybe inhibit the former mainly then fulfill the function of anti-hepaticfibrosis.