丹参川芎嗪注射液对脓毒症幼鼠急性肝损伤保护作用的研究

Protective Effect of Danshen Chuangxiongqin Injection on Acute Liver Injury in Septic Young Rats Based on Nrf2/HO-1 Pathway

目的探讨丹参川芎嗪注射液对脓毒症幼鼠急性肝损伤的保护作用及其可能机制。方法将36只SD幼鼠随机分为3组:对照组、模型组、丹参川芎嗪注射液组,各12只。采用腹腔注射脂多糖(LPS)10mg/kg建立脓毒症幼鼠模型,丹参川芎嗪注射液组1h后给予腹腔注射丹参川芎嗪注射液5mL,分别于造模后6、12h各取6只幼鼠采集血样和肝组织样品,光镜下观察肝组织病理改变,检测血清谷草转氨酶(AST)、谷丙转氨酶(GOT)和乳酸脱氢酶(LDH)水平和肝组织中丙二醛(MDA)、谷胱甘肽(GSH)水平及超氧化合物歧化酶(SOD)、谷胱甘肽硫转移酶(GST)活性,采用qRT-PCR和Western blotting检测肝组织中核转录因子E2相关因子2(Nrf2)和血红素氧化酶1(HO-1)mRNA和蛋白表达水平。结果对照组肝细胞排列整齐,形态正常,结构完整无病理改变;模型组幼鼠6h时肝细胞出现水肿、部分坏死,12h时水肿加重,部分肝细胞颗粒变性,细胞排列紊乱;丹参川芎嗪注射液组6、12h肝细胞水肿、坏死、颗粒变性与模型组比较明显减轻;与对照组比较,模型组6、12h血清AST、GOT和LDH水平及肝组织MDA水平均升高(P<0.05),肝组织GSH水平及SOD、GST活性均降低(P<0.05);与模型组比较,丹参川芎嗪注射液组6、12h血清AST、GOT和LDH水平及肝组织MDA水平均降低(P<0.05),肝组织GSH水平及SOD、GST活性均升高(P<0.05);与对照组比较,模型组肝组织中Nrf2、HO-1mRNA和蛋白均升高(P<0.05);与模型组比较,丹参川芎嗪注射液组肝组织中Nrf2、HO-1mRNA和蛋白均升高(P<0.05)。结论丹参川芎嗪注射液对脓毒症致急性肝损伤具有保护作用,其保护机制可能与激活Nrf2/HO-1通路,增强肝组织抗氧化能力有关。

Objective:To investigate the protective effect of Danshen Chuangxiongqin Injection on acute liver injury in sepsis young rats and its possible mechanism.Methods:36 SD young rats were randomly divided into three groups:the control group,the model group and Danshen Chuangxiongqin Injection group,12 rats in each. The sepsis young rats model was established by intraperitoneal injection of LPS 10 mg/kg.Blood samples and liver tissue samples were collected from 6 young rats at 6 and 12 hours after the establishment of the model respectively;the pathological changes of liver tissues were observed under light microscope;the levels of serum AST,GOT, LDH,MDA,GSH,SOD and GST in liver tissues were measured’the expression levels of Nrf2 and HO-1 in liver tissues were detected by qRT-PCR and Western blot.Results:The hepatocytes in the control group were well arranged,normal in shape and intact in structure,without pathological changes;hepatocytes in the model group appeared edema and necrosis at 6 h,edema aggravated at 12 h,some hepatocyte granules showed degeneration and arranged disorderly;hepatocytes in the Danshen Chuangxiongqin Injection appeared edema and necrosis at 6 h and 12 h,the granule denaturation was significantly reduced compared with the model group;compared with the control group,the levels of AST,GOT,and LDH in serum and MDA in liver tissues of the model group increased at 6 h and 12 h(P < 0.05),GSH level and SOD,GST activities in liver tissues were decreased(P < 0.05);compared with the model group,the levels of AST,GOT,and LDH in serum and MDA in liver tissues of Danshen Chuangxiongqin Injection group decreased at 6 h and 12 h(P < 0.05),the levels of GSH and activities of SOD and GST in liver tissues were increased(P < 0.05);compared with the control group,the expressions of Nrf2 and HO-1 mRNAs and proteins in the liver tissues of the model group increased(P < 0.05);compared with model group,the expressions of Nrf2 and HO-1 mRNAs and proteins in liver tissues of Danshen Chuangxiongqin Injection group increased(P < 0.05).Conclusion:Danshen Chuangxiongqin Injection has a protective effect on acute liver injury induced by sepsis,and the protective mechanism may be related to activating Nrf2/HO-1 pathway and enhancing the antioxidant capacity of liver tissues.