双虎清肝颗粒对四氯化碳诱发肝纤维化大鼠血液TNF-α和肝脏组织学的影响

Effects of Shuanghu Qinggan Granule on Hepatic Expression of TNF-α and Pathological Changes of Hepatic Fibrosis Induced by Carbon Tetrachloride in Rats

目的:探讨双虎清肝颗粒对四氯化碳诱发肝纤维化大鼠血液TNF-α和肝脏组织学的影响。方法:清洁级Sprague-Dawley大鼠60只,随机分成6组,A组(正常对照组)、B组(模型组)、C1组(双虎清肝颗粒小剂量组)、C2组(双虎清肝颗粒中剂量组)、C3组(双虎清肝颗粒大剂量组)、D组(水飞蓟素对照组)。除正常对照组外,均经皮下注射400mL/L四氯化碳8周,后各治疗组给予不同剂量双虎清肝颗粒干预8周,阳性对照组给予水飞蓟素干预8周,ELISA法检测肿瘤坏死因子(TNF-α),常规HE及Masson染色,光镜观察肝组织的脂肪变、炎症及纤维化程度。结果:各组大鼠肝脏病理表现:A组肝细胞排列整齐,未见脂质空泡形成,汇管区未见炎症,肝小叶结构清晰可见,未见肝纤维化及假小叶形成;B组可见大量脂质空泡形成,汇管区可见大量炎性细胞浸润,成纤维细胞大量增生,肝细胞广泛变性呈片状坏死。汇管区可见大量假小叶形成,肝小叶间隔明显增宽和大量胶原纤维沉积;C1、C2、C3、D组:C1组同B组相比,未见明显差异,可见大量脂质空泡形成,汇管区见大量炎性细胞浸润、成纤维细胞增生,肝细胞广泛变性及点状、片状坏死及较多假小叶形成。C2组同模型组相比,脂质空泡有所减少,肝细胞坏死程度有所减轻,假小叶形成较少,仍明显可见、较多胶原纤维沉积。C3、D组同B组相比,脂质空泡明显减少,肝细胞坏死程度明显减轻,假小叶形成、胶原纤维沉积明显减少(见图1)。各组大鼠肝纤维化病变半定量结果:A组大鼠纤维化程度为0,B组大鼠纤维化程度为16.1±5.5,C1组大鼠纤维化程度为12.6±4.6a,C2组大鼠纤维化程度为10.7±3.9a,C2组大鼠纤维化程度为6.4±3.4a,D组大鼠纤维化程度为6.5±3.3a,以上数据为半定量计数结果,计算公式:P+L+2×(PN+BN);与B组比较,aP<0.05。各组大鼠肝脏TNF-α表达结果:A组大鼠8.01±3.45b,B组大鼠19.15±4.86,C1组大鼠11.28±3.43a,C2组大鼠9.93±5.31a,C3组大鼠9.37±5.02a,D组大鼠10.86±5.28a。与B组比较,aP<0.05,与B组比较,bP<0.01。结论:双虎清肝颗粒能够显著改善四氯化碳所诱发的肝纤维化大鼠肝脏组织学变化及TNF-α的表达,并且量效关系明显。

Objective： To investigate the effects of Shuanghu qinggan granule on hepatic expression of TNF-α and pathological changes of hepatic fibrosis induced by carbon tetrachloride in rats.Methods ：60 Sprague-Dawley rats were divided into 6 groups,namely normal control group（group A）,model group（group B）,Shuanghu qinggan granule small,middle,large dose groups（groupC1,C2,C3） and silymarin contrast group（group D）. The rats group B,group C1,C2,C3 and group D were subcutaneously injected with carbon tetrachloride （3mL per kilogram body weight）twice a week for 8 weeks. Then the rats of groupC1,C2,C3 were administered with Shuanghu qinggan granule at different dose for 8 weeks,including 1.2 grams per kilogram body weight （groupC1）,2.5 grams per kilogram body weight （groupC2）,5 grams per kilogram body weight（groupC3）.The rats of group D were then administered with silymarin at the dose of 50 milligram per kilogram body weight. The levels of serum TNF-α of all rats were measured respectively under th...更多e same condition and at the same time by elisa. The pathological changes of hepatic fibrosis and the density of collagen in liver specimens of all rats were assessed through HE and Masson staining.Results ：Compared with group B, lipid vacuoles and liver necrosis were significantly reduced, and false lobular formation of collagen deposition was significantly decreased in group C3 and D. There was fibrosis difference between group C1, C2, C3 and group D （12.6 ± 4.6, 10.7 ±3.9, 6.4 ± 3.4, 6.5 ± 3.3 vs 16.1±5.5, all P 〈 0.05）. Serum TNF-α levels were lower in group C1,C2,C3, D than in group B （19.15±4.86,11.28±3.43, 9.93±5.31,9.37±5.02,10.86±5.28 vs 8.01±3.45, P〈0.001 or 0.05）. Conclusion ：Shuanghu qinggan granule has obvious effects on hepatic expression of TNF-α and pathological changes of hepatic fibrosis induced by carbon tetrachloride in rats. Significant difference was found in fibrosis values,and the higher dosage of Shuanghu qinggan granule was more effective.