截断逆挽方对慢加急性肝衰竭模型大鼠血清TNF-α、肝组织p-JNK及c-Jun的影响  

Effects of Jieduan Niwan Formula on the c-Jun,p-JNK of liver tissue in Acute-on-Chronic liver failure rats

作　　者：穆凌云[1] 李金霞[1] 张秋云[1] 陈煜[2] 高连印[1] 杜宇琼[1] Mu Lingyun Li Jinxia Zhang Qiuyun Chen Yu Gao Lianyin Du Yuqiong(Traditional Chinese Medicine Academy of Capital Medicine University,Beijing Key Laboratory of Traditional Chinese Medicine Collateral Disease Research ,Beijing 100069, China 3. Artificial Liver Center, Beijing Yonan Hospital, Capital Medical University, Beijing 100069, Chin)

机构地区：[1]首都医科大学中医药学院中医络病研究北京市重点实验室,北京100069 [2]首都医科大学附属北京佑安医院人工肝中心,北京100069

出　　处：《首都医科大学学报》2017年第2期282-288,共7页Journal of Capital Medical University

基　　金：首都中医药研究专项(14ZY01)~~

摘　　要：目的观察截断逆挽方对慢加急性肝衰竭(acute-on-chronic liver failure,ACLF)大鼠血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量、肝组织磷酸化c-Jun氨基末端激酶(phosphorylated c-Jun N-terminal kinase,p-JNK)及转录因子cJun表达的影响,探讨该方干预ACLF大鼠的部分作用机制。方法选择SPF级Wistar雄性大鼠70只,采用数字表法随机分为正常对照组、模型组、截断逆挽方组和JNK抑制剂SP600125(anthrapyrazolone)组,予猪血清免疫诱导大鼠形成肝纤维化模型,再联合D-氨基半乳糖/脂多糖(D-galactosamine/lipopolysaccharide,D-Gal N/LPS)急性攻击,建立ACLF大鼠模型。截断逆挽方组在急性攻击前给予截断逆挽方连续灌胃3 d,SP600125组在急性攻击前0.5 h腹腔注射SP600125。各组大鼠分别在急性攻击后4、8、12 h取材。采用酶联免疫吸附法(enzyme-linked immune sorbent assay,ELISA)检测血清TNF-α含量,蛋白印迹法(Western blotting,WB)检测c-Jun蛋白表达量,免疫组织化学法检测肝组织中p-JNK蛋白表达量,HE染色观察肝脏结构病理变化。结果与正常组相比,模型组各时间点血清TNF-α浓度均升高,差异具有统计学意义(P<0.01);与对应时间点模型组相比,截断逆挽方组及SP600125组4、8 h血清TNF-α含量明显降低(P<0.01),12 h有所升高;与正常组相比,模型组各时间点c-Jun、p-JNK表达量升高(P<0.01);与模型组相比,截断逆挽方组、SP600125组在4 h及8 h c-Jun、p-JNK表达减少(P<0.05),12 h表达增多。截断逆挽方组及SP600125组两两比较差异无统计学意义(P>0.05)。结论截断逆挽方在一定程度上可以减轻ACLF大鼠肝细胞的损伤,其作用机制可能是通过影响JNK信号通路介导的细胞凋亡,从而保护肝细胞,抑制肝衰竭。Objective To observe the effect of the Jieduan-Niwan Formula （JDNW）on the serum tumor necrosis factor alpha（TNF-α）content,the expression of p-JNK and transcription factor c-Jun in acute-on-chronic liver failure（ACLF）rats,to discuss part of the action mechanism of intervention of in ACLF.Methods SPF 70 male Wistar rats were randomly divided into normal control group,model group,JDNW group and JNK inhibitor SP600125 group,to inject pig serum to induce rat liver fibrosis model,and then acute attack combined with D-GalN/LPS,ACLF model was established.Rats in JDNW group were orally given JDNW formula for 3 days before acute attack.Rats in SP600125 group giving intraperitoneal injection half an hour before acute attack.Rats were sacrificed,respectively at 4,8 and 12 h after model established.The expression of TNF-α,c-Jun,p-JNK were detected by enzyme-linked immune sorbent assay（ELISA）,western blot method and immunohistochemical analysis,respectively.Results Compared with normal group,the expression of TNF-α,p-JNK,c-Jun of model group were gradually increased;compared with the model group,the expression of TNF-α,p-JNK,c-Jun of JDNW and SP600125 group in 4h and 8h was less（P〈0.05）,and the expression of 12h increased;the different expression of SP600125 group was in accordance with the JDNW group.Conclusion The prescription of JDNWcan reduce the damage of rat liver cell ACLF in a certain extent,its mechanism may be through cell apoptosis mediated by JNK signaling pathway,thereby protecting liver cells,inhibiting liver failure.

关 键 词：慢加急性肝衰竭 截断逆挽方 细胞凋亡 JNK信号通路 

分 类 号：R285.6[医药卫生—中药学]