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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。
OBJECTIVE: To understand the effect of Ling Jia Fang on prevention and treatment of liver fibrosis in animal models of blood stasis-induced liver fibrosis and to explore its mechanism of prevention and treatment of liver fibrosis from the aspects of protein expression of OPN and PAI-1. Methods: A total of 70 Wistar rats were randomly divided into 4 groups: normal control group, Ling Jia Fang group, Fufang Biejia Ruangan Tablet group, blood stasis group. Except for the normal control group, the other three groups were injected with DMN via tail vein and bovine serum albumin and norepinephrine were injected into the other three groups. The rat model of hepatic fibrosis with blood stasis was duplicated for 4 weeks. After 4 weeks Lingjiafang group Lingling prescription gavage once a day for a total of 8 weeks. Fufang Biejia Ruangan Tablet 4 weeks after the control group Fufang Biejia Ruangan Tablet gavage once a day for a total of 8 weeks. At the end of the 8th week, blood was collected to detect the levels of ALT, AST, Alb and Glb. The liver tissues were harvested for protein expression of OPN and PAI-1 by Western blot. Results: Compared with the blood stasis group, the ALT of Ling Jia Fang and Fufang Biejia Ruangan Pian in rats decreased significantly (P <0.05), and the AST in Lingjia Recipe decreased significantly (P <0.01) ; Compared with blood stasis group, the body weight of rats treated with Ling Jia Fang increased significantly (P <0.01); the body weight of rats treated with Fufang Biejia Ruangan Pian also increased (P <0.05); Compared with that of blood stasis group (P <0.01), and the ratio of spleen to body weight decreased (P <0.05), and the ratio of liver to body in Ling Jia Fang was significantly higher than that of Fufang Biejia Ruangan Pian in treatment group Increased more significantly (P <0.05). The expression of OPN and PAI-1 in Ling Jia Fang was significantly lower than that in blood stasis group (P <0.01), and Ling Jia Fang was more effective than Fufang Biejia Ruangan Tablet in inhibiting OPN expression Significance (P <0.05). CONCLUSION: Ling Jia Recipe may indirectly decrease PAI-1 protein expression by inhibiting the expression of OPN, so it appears that the decrease of PAI-1 protein expression is greater than that of OPN, so that inhibition of extracellular matrix degradation by PAI-1 Weakened role, increase the degradation of extracellular matrix, exert anti-fibrosis.