柔肝抑纤饮联合二至丸逆转DMN大鼠肝窦毛细血管化的对比研究

The Experimental Study on anti Sinusoid Capillarization of Rougan Yixianyin Decoction Combined with Erzhi Pill

目的:探讨中药复方柔肝抑纤饮联合二至丸在抗大鼠肝窦毛细血管化方面与单独使用柔肝抑纤饮有无差别,明确联合二至丸后对原方有无增效、协同作用,并阐明其机制。方法:建立大鼠DMN肝纤维化模型,随机分4组:A组(正常组)、B组(模型组)、C组(柔肝抑纤饮组)、D组(柔肝抑纤饮+二至丸组),分别灌服相应药物。检测血清HA、col-Ⅳ,行胶原染色(M asson),免疫组化染色观察并测定肝组织CD44、vWF、MMP-2、MMP-9的表达,电镜观察窦周隙超微结构。结果:D组与C组在降低HA含量、M asson染色百分比以及改善CD44和MMP-9免疫组化染色阳性面积均值等方面均明显优于B组(P<0.05,P<0.01),但D组与C组比较未见显著性差异(P>0.05)。D组在降低col-Ⅳ、MMP-2和vWF免疫组化染色阳性面积均值等方面,作用明显优于C组(P<0.05);电镜观察发现,D组肝窦内皮失窗孔和内皮下连续性基底膜的形成等病变也相对较轻。结论:联合二至丸能使柔肝抑纤饮抗肝窦毛细血管化的作用得以增强,提高疗效。

Objective ： Try to find the difference of the effects in preventing the form of sinusoid capillartzation between Rougan Yixianyin Decoction only and the Decoction combined with Erzhi Pill, thus reveal the mechanism and the synergistic action. Methods ： The earlier stage of rat hepatic fibrosis model was made by DMN with peritoneal injection, 48 rats were divided into 4 groups： group A （Normal control group）, group B （Model group）, group C （Rougan Yixianyin group）, group E （ Rougan Yixianyin ＋ Erzhi - pill group）, Rats of each group were infected with DMN except group A. After 4 weeks of injection, group C were fed with Rougan Yixianyin Decoction, group D with Decoction of Rougan Yixia- nyin ＋ Erzhi pill, and group A, B were fed with physiological saline respectively for 14 days. Rats of each group were put into death, tested the contents of HA and col - 1V in serum. The histopathology of hepatic tissue was observed too （by Masson dye, electron microscope, and immunohistochemical methods）, The expression of CD64, vWF, MMP - 2, MMP - 9 were measured by quantitative analysis. Results ： In terms of lower HA in serum and the lower masculine area of collagen （masson）, CD44 and MMP -9, there were no significant difference between Rougan Yixianyin group and Rougan Yixianyin ＋ Erzhi pill group（P 〉0.05）, but there were more significant difference than that in model group（ P 〈 0.05, P 〈0.01 ）. In terms of lower content of col - IV in serum,lower masculine area of mmp -2 and vWF in hepatic tissue, there were significant difference between Rougan Yixianyin group and Rougan Yixianyin ＋ Erzhi pill group（ P 〈0.05）, also the damage of sinusoid - lining endothelium cells and basal lamina in Rougan Yixianyin ＋ Erzhi pill group were no more obviously than that in Rougan Yixianyin group, so the whole curative effect of Rougan Yixianyin ＋ Erzhi pill was better than Rougan Yixianyin. Conclusions： Rougan Yixianyin combined with Erzhi Pill has synergistic action to resist he- patic fibrosis and sinusoid capillartzation, so more high curative effect can be elevated.