健脾软肝方对CCL_4诱导的肝纤维化大鼠肝窦内皮细胞失窗孔的影响  

The Effect of Jianpi Ruan'gan Decoction (健脾软肝方) on the Fenestra of Liver Hepatic Sinusoidal Endothelial Cells in Rats with Hepatic Fibrosis Induced by CCL4

作　　者：陈文玲[1] 陈文慧[1] 杜良 朱晓松[1] 姚政[1] 石安华[1] CHEN Wen-ling;CHEN Wen-hui;DU Ling;ZHU Xiao-song;YAO Zheng;SHI An-hua(Yunnan University of Traditional Chinese Medicine,Kunming Yunnan 650011,China;Ninth People＇s Hospital of Benxi,Benxi Liaoning 117000,China)

机构地区：[1]云南中医学院,云南昆明650011 [2]本溪市第九人民医院,辽宁本溪117000

出　　处：《中医药导报》2018年第17期13-18,30,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：云南省应用基础研究计划(2013FZ091)

摘　　要：目的:探讨健脾软肝方对CCL_4诱导的肝纤维化大鼠肝窦内皮细胞失窗孔的影响。方法:60只雄性Wistar大鼠随机分为6组,采用2 m L/kg的30%CCL_4与70%橄榄油混合液腹腔注射,2次/周,复制肝纤维化模型。造模同时给予药物干预,正常组和模型组均给予生理盐水灌胃,阳性对照组用扶正化瘀方灌胃,健脾软肝方高、中、低剂量组分别给予不同剂量的健脾软肝方灌胃。4周后,采用HE染色和Mallory染色观察肝组织纤维化程度,透射电镜观察肝窦内皮细胞窗孔变化,ELISA法检测血清HA含量、Real-time PCR及免疫组化ABC法检测CD31、CD44mRNA及蛋白表达。结果:健脾软肝方干预后,光镜显示肝纤维化程度比模型组减轻;电镜显示大鼠肝窦内皮细胞窗孔直径变大、数目增多;HA含量降低(P<0.01),且高、中、低剂量组血清HA的降低随剂量的升高而变化,表现为一定的剂量依赖关系;CD44阳性及mRNA表达增高(P<0.05),CD31阳性及mRNA表达降低(P<0.05),且表达量随剂量的升高而逐渐降低,呈一定的剂量依赖关系。结论:健脾软肝方可减轻肝纤维化大鼠LSEC失窗孔的作用。Objective： To investigate the intervention effect of Jianpi Ruan＇gan Decoction on the fenestra of liver sinusoidal endothelial cells in rats with hepatic fibrosis induced by CCL4. Methods： A total of 60 male Wistar rats were randomly divided into 6 groups by random number table method. Experimental model of hepatic fibrosis was induced by injected with intraperitoneal injection of 30% CCL4 and 70% olive oil mixture, 2 m L/kg, 2 times per week. Drug intervention was taken at the same time. The model group and the normal group were given normal saline, the positive control group with Fuzheng Huayu Decoction（扶正化瘀方）, and the Jianpi Ruan＇gan Decoction of high, medium and low dose group were given different doses of Jianpi Ruan＇gan Decoction orally. After 4 weeks, t the fibrosis degree of liver tissue was observed by HE staining and Mallory staining. The changes of the fenestra of liver hepatic sinusoidal endothelial cells were observed by transmission electron microscope. The serum HA content, Real-time PCR and immunohistochemical ABC method were used to detect CD31, CD44 mRNA and protein expression by ELISA method. Results： After the intervention of Jianpi Ruan＇gan Decoction, the degree of hepatic fibrosis was lower than that of the model group by light microscope,the diameter and number of the fenestra of liver sinusoidal endothelial cells were increased. The content of HA was decreased（P〈0.01）, and the decrease of serum HA in high, middle and low dose groups changed with the increase of dose, which showed a dose-dependent relationship. The expression of CD44 positive and mRNA was increased（P〈0.05）, the expression of CD31 positive and mRNA was decreased（P0.05）, and the expression level gradually decreased with the increase of dose, which showed a dose-dependent relationship. Conclusion： Jianpi Ruan＇gan Decoction can reduce the function of the fenestra of liver sinusoidal endothelial cells in rat liver fibrosis.

关 键 词：肝纤维化 健脾软肝方 肝窦内皮细胞 窗孔 

分 类 号：R285.5[医药卫生—中药学]