大黄赤芍注射液对急性肝衰竭大鼠治疗作用及机制探讨

Therapeutic Effect and Mechanism of Dahuang Chishao Injection(大黄赤芍注射液) on Rats with Acute Liver Failure

目的观察大黄赤芍注射液对肝衰竭大鼠炎症因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)]及抑制肝细胞凋亡因子[C-X3-C趋化因子配体1(CX3CL1)、B细胞淋巴瘤/白血病基因-2(BCL-2)、细胞周期蛋白D1(CyclinD1)]表达变化的影响。方法将2月龄大鼠随机分为空白组,造模阳性对照组,大黄赤芍注射液低、中、高剂量组及促肝细胞生长素组。除空白组外,其余各组采用D-氨基半乳糖(D-Gal)800 mg/kg,脂多糖(LPS)8μg/kg腹腔注射,制造急性肝衰竭模型。造模前3 d至实验结束大黄赤芍注射液低、中、高剂量组分别以大黄赤芍注射液低、中、高剂量[0.25、0.5、0.75 mL/(100 g·d)]尾静脉注射给药(1次/d);空白组及造模阳性对照组均予0.9%氯化钠注射液1 mL/(100 g·d);促肝细胞生长素组参照文献给予促肝细胞生长素10 mL/(kg·d)。造模48 h后处死大鼠取肝组织病理标本,HE染色后在光镜下观察肝组织的病理改变,采用qPCR、Western Blot技术验证不同组间的TNF-α、IL-6、CX3CL1、BCL-2、CyclinD1是否存在表达差异。结果药物干预组可缓解肝组织病理损伤,同时降低了TNF-α、IL-6表达量,提高了CX3CL1、BCL-2、CyclinD1表达量,与对照组比较差异具有统计学意义(P<0.05)。结论大黄赤芍注射液对肝衰竭的疗效作用机制可能与减轻炎性反应、抗肝细胞凋亡、促进肝细胞再生相关。

Objective To observe the effects of Dahuang Chishao Injection(大黄赤芍注射液)on inflammatory factors(TNF-α,IL-6)and inhibiting hepatocyte apoptosis factors(CX3CL1,BCL-2,CyclinD1)expression changes in liver failure rats.Methods Two month old rats were randomly divided into blank group,positive control group,Dahuang Chishao Injection low,medium and high dose groups and hepatocyte growth promoting hormone group.Except for the blank group,acute liver failure model was established by intraperitoneal injection of D-Gal 800 mg/kg and LPS 8μg/kg.From 3 days before modeling to the end of the experiment,low,medium and high doses of Dahuang Chishao Injection[0.25,0.5,0.75 m L/(100 g·d)]were injected into the tail vein(once a day).The blank group and positive control group were given 0.9% normal saline 1 m L/(100 g·d),and the hepatocellular auxin promoting group was given 10 m L/(kg·d)according to the literature.48 h after modeling,the liver tissue was sacrificed and pathological samples were collected.After HE staining,the pathological changes of liver tissue were observed under light microscope.qPCR and Western Blot were used to verify whether there were differences in the expressions of TNF-α,IL-6,CX3 CL1,BCL-2 and Cyclin D1 between different groups.Results The pathological injury of liver tissue was alleviated in the drug intervention group,while the expression levels of TNF-α and IL-6 were decreased,and the expression levels of CX3 CL1,BCL-2 and Cyclin D1 were increased in the drug intervention group compared with the control group,and the differences were statistically significant(P<0.05).Conclusion The mechanism of Dahuang Chishao Injection on liver failure may be related to reducing inflammatory reaction,resisting hepatocyte apoptosis and promoting hepatocyte regeneration.