大黄灵仙颗粒调控IL-6、EGR-1表达治疗胆石症的实验研究

Dahuang Lingxian Granule Treating Cholelithiasis by Down-regulating IL-6 and EGR-1

目的:明确大黄灵仙颗粒对慢性肝损伤胆石病兔模型的疗效,研究中药复方对关键致石因素IL-6、EGR-1表达的调控作用。方法:(1)将50只雄性新西兰大白兔随机分为3组:正常对照组、肝纤维化胆石病组、肝硬化胆石病组,其中正常对照组10只,其余两组各20只,在金博的慢性肝损伤兔模型构建方法的基础上,同时予高热量、高胆固醇致石饲料,建立慢性肝损伤胆石病兔模型。(2)造模12周后,病理明确诊断肝损伤形成后,将各组剩余实验兔分别分为对照组、治疗组,其中治疗组每天灌胃中药制剂大黄灵仙颗粒0.4 g/(kg·d),其余各组灌胃等量生理盐水,各组大白兔均正常饲料饲养。用药第13周,麻醉后取肝脏组织,分别使用实时荧光定量PCR、免疫组化等方法进行检测。结果:(1)常规病理:与对照组相比,用药各组光镜下可见遭到破坏的肝小叶逐渐修复,部分肝细胞形态恢复正常,炎性细胞及纤维组织减少,区域呈点状、片状坏死明显减少;(2)免疫组化:与对照组相比,喂食大黄灵仙颗粒剂的各组肝窦、汇管区的肝细胞胞浆或胞膜上棕褐色颗粒沉积表达明显减少,小胆管胆汁淤积减少,说明用药后肝损伤得到改善;(3)q PCR检测结果:与对照各组相比,用药各组的IL-6、EGR-1 mRNA的表达量均有不同程度降低(P<0.05),说明该药可有效减轻肝损伤。结论:本研究结果表明大黄灵仙颗粒对慢性肝损伤性胆石病动物模型具有较明显的改善和防治作用。

Objective：To validate the effects of Dahuang Lingxian Granule on liver fibrosis cholelithiasis in rabbit model and elucidate the molecular mechanism of Dahuang Lingxian Granule regulating the expression levels of IL-6 and EGR-1.Methods：（1）Fifty male New Zealand rabbits were divided into 3 groups randomly：liver fibrosis group（20 rabbits）,liver fibrosis cholelithiasis group（20 rabbits） and normal control group（10 rabbits）.The rabbit model with liver fibrosis cholelithiasis was established as previously described.（2）After the existence of the hepatic fibrosis was confirmed,the remaining rabbits in each group were divided into the control group and the treatment group.The treatment group were treated with 0.4 g/（kg·d） Dahuang Lingxian Granules by gastric irrigation and the other groups were treated with equal doses of normal saline and all rabbits were on normal diet.In the 13 th week,liver tissues of all the rabbits were tested by PCR and immunochemistry staining.Results：（1）General pathology：Compared with the control group,the destruction liver tissue was repaired gradually and the arrangement of liver cells changed slightly.Inflammatory cells and fibrous tissue reduced.Less spotty or patchy necrosis were observed in treatment group.（2） Immunochemistry staining：In comparison with control group,less brown particles were observed in the cytoplasm and membrane of live cells in sinusoids and hepatic duct area and cholestasia in small bile ducts less frequently happened,suggesting the positive effects of Dahuang Lingxian Granule on liver damage.（3） Results of PCR：The mRNA levels of IL-6 and EGR-1 in groups treated with Dahuang Lingxian Granule were decreased than those of control group（P〈0.05）,which indicates that this drug could effectively reduce the live damage.Conclusion：This study showed that Dahuang Lingxian Granule could attenuate liver fibrosis cholelithiasis in rabbit model.