壮肝逐瘀煎对纤维化大鼠肝脏微循环障碍的调控

Microcirculatorydisorder is the lifeblood of pathological basis and condition of hepatic fibrosis in chronic liver disease of various progress,portal hypertension and even cancer"zhuang gan Zhu yu" Decoction has been confirmed to regulate the secretion of vasoactive substances and the hemodynamics of portal hypertension of liver cirrhosis,The hepatofibrosis rats model was induced by CCL4 compound factors,and treated with "zhuang gan Zhu yu"Decoction and the different combinations.Hepatic microcirculation,microvascular blood flow velocity and blood flow was assayed by contrast-enhanced ultrasoundgraphy.The ultrastructure of hepatic tissue was assayed by electron microscope,Vasoactive substance（ET-1/NO/iNOS）were assayed by ELISA and the method of nitric acid reductase.The mRNAs of promoting/inhibiting angiogenesis factor(TSP-1/VEGF/ PDGF/ Ang-1,2/HIF-1a) and receptors(ET-A/BR/Tie-2/VEGFR-1,2/PDGFR) were quantified by real-time RT-PCR.The number of MVD and arteriole of liver tissue were assayed by Immunohisto-chemical staining,The microcirculatory regulation revealed anti-hepatic fibrosis action mechanism and rules of "zhuang gan Zhu yu" Decoction and different treatment/prescription,griped the lifeblood of fibrosis progression to cirrhosis,portal hypertension,the progress of the disease and cancer,and provided the reference for anti-hepatic fibrosis of the liver and other organ,the research of improve microcirculation.

微循环障碍是各种慢性肝病肝纤维化的病理基础及病情进展、门脉高压发生甚至癌变的"命脉"。临床证实壮肝逐瘀煎能调节血管活性物质分泌及改善肝硬化门脉高压血流动力学，本研究通过复制CCL4复合因素大鼠肝纤维化模型，给予壮肝逐瘀煎及不同组合药液干预，应用超声微泡造影评估肝脏微循环、微血管血流状态和速度，采用电镜观察肝组织超微结构、ELISA及硝酸还原酶法检测血管活性物质（ET-1、NO、iNOS）活性、RT-PCR及免疫组化染色分别检测促/抑血管新生因子（TSP-1、VEGF、PDGF、Ang-1，2、HIF-1a）基因表达及受体（ET-A，BR、Tie-2、VEGFR-1，2、PDGFR）分布差异、肝组织MVD和小动脉数量，围绕微循环调控揭示壮肝逐瘀煎及不同治法/组方抗肝纤维化的作用机制及规律，扼住纤维化病情进展、门脉高压发生甚至癌变的"命脉"，为肝脏及其它脏器纤维化防治、改善微循环研究提供参考。

肝纤维化是各种慢性肝病向肝硬化发展的必经途径，微循环障碍是各种慢性肝病肝纤维化的病理基础及病情进展、门脉高压发生甚至癌变的“命脉”。临床证实壮肝逐瘀煎能调节血管活性物质分泌及改善肝硬化门脉高压血流动力学，本研究通过复制CCL4复合因素大鼠肝纤维化模型，给予壮肝逐瘀煎及不同组合药液干预，应用超声微泡造影评估肝脏微循环、微血管血流状态和速度，采用电镜观察肝组织超微结构，ELISA及硝酸还原酶法检测血管活性物质（ET-1、NO、iNOS）活性，RT-PCR及免疫组化染色分别检测促/抑血管新生因子（TSP-1、VEGF、PDGF、Ang-1，2、HIF-1a）基因表达及受体（ET-A，BR、Tie-2、VEGFR-1，2、PDGFR）分布差异、肝组织MVD和小动脉数量，围绕微循环调控揭示壮肝逐瘀煎及不同治法/组方抗肝纤维化的作用机制及规律，扼住纤维化病情进展、门脉高压发生甚至癌变的"命脉"，为肝脏及其它脏器纤维化防治、改善微循环研究提供参考。通过实验发现：①C组及不同组合分组和G组均能不同程度改善肝纤维化大鼠肝脏病理组织、减轻大鼠肝纤维化的程度，与G组相比，有统计学意义；②肝纤维化大鼠造模成功后，ET-1和NO血清含量B组较A组高（P＜0.05）；药物治疗后，C、D、E、F、G组与B组比较，血清中的ET-1和NO血清含量明显降低（P＜0.05），其中C组和G组比较，壮肝逐瘀煎全方组对肝纤维改善较为明显（P＜0.05），表明壮肝逐瘀煎能够降低血清中ET-1和NO的浓度，阻止肝纤维化的进展。③壮肝逐瘀煎可降低VEGE及PDGF的基因表达量，减少肝纤维化过程中肝脏组织微血管的再生速度及再生程度，改善微循环，达到抗肝纤维化的作用。④C组及不同组合分组均能够改善造影剂到达肝脏中的各项指标，但壮肝逐瘀煎全方组效果更显著，提示壮肝逐瘀煎可改善纤维化大鼠肝脏微循环。根据实验结果，可以推测壮肝逐瘀煎可以改善肝脏微循环而起到抗肝纤维化作用。本研究以肝脏微循环为切入点，探讨了壮肝逐瘀煎及不同治法/组方抗肝纤维化的作用机制及规律，为临床上延缓甚至逆转肝纤维化提供依据。