Delta阿片受体激活通过调节JAK2/STAT3通路减轻肝硬化大鼠缺血再灌注损伤  

Protective effect of delta opioid receptor activation on hepatic ischemia reperfusion injury via JAK2/STAT3 pathway in rats with cirrhosis

作　　者：刘珏莹[1] 潘钱玲 殷文渊[1] 王苑[1] 俞卫锋[1] LIU Jueying;PAN Qianling;YIN Wenyuan;WANG Yuan;YU Weif?eng(Department of Anesthesiology,Renji Hospital Affiliated to School of Medicine of Shanghai Jiaotong University,Shanghai 200127,China)

机构地区：[1]上海交通大学医学院附属仁济医院麻醉科

出　　处：《实用医学杂志》2019年第22期3441-3446,共6页The Journal of Practical Medicine

基　　金：国家自然科学基金青年项目(编号:81400644)

摘　　要：目的研究Delta阿片受体激活对肝硬化大鼠肝脏缺血再灌注(ischemia reperfusion,IR)损伤的保护作用以及相关机制。方法选择雄性健康Sprague-Dawley大鼠,建立肝硬化模型。随后32只大鼠随机分为4组(n=8),分别为假手术组(SHAM组)、对照组(CON组)、Delta阿片受体激动剂预处理组(DADLE组)、Delta阿片受体拮抗剂组(NTD组)。再灌注6 h后,通过H&E染色评分,测血清丙氨酸氨基转移酶(alanine transaminase,ALT)和天冬氨酸氨基转移酶(aspartate transaminase,AST)水平。使用酶联免疫吸附试验检测血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)以及白介素-1β(interleukin-1β,IL-1β)的含量。采用荧光定量反转录-聚合酶链反应以及蛋白印迹法检测Janus激酶2(Janus kinase 2,JAK2)和信号转导及转录激活因子-3(signal transducer and activators of transcription 3,STAT3)的表达。结果与CON组相比,DADLE明显降低血清ALT[(138±33.369)U/L vs.(267±106.173)U/L,P <0.05]和AST[(210±102.666)U/L vs.(374±64.021)U/L,P <0.05]。DADLE组H&E染色评分明显低于CON组(4.83±1.169 vs.7.67±0.816,P <0.05)。与CON组相比,DADLE明显降低了TNF-α[(15.561±5.12)pg/mL vs.(32.289±9.23)pg/mL,P <0.05]和IL-1β的含量[(46.444±7.777)pg/mL vs.(68.841±9.225)pg/mL,P <0.05]。使用DADLE后,JAK2和STAT3的mRNA表达增高(P <0.05),磷酸化蛋白含量也明显升高。然而NTD组与CON组相比差异无统计学意义(P> 0.05)。结论 Delta阿片受体激活能减轻硬化肝脏IR损伤,其机制可能与调节JAK2/STAT3通路有关。Objective To investigate the effect of DALTA opioid receptor activation on cirrhotic rats′ livers ischemia reperfusion(IR)injury and related mechanism. Methods Male Sprague-Dawley rats were established liver cirrhosis model. 32 rats with liver cirrhosis were randomly divided into 4 groups(n = 8):SHAM group;CON group;Delta opioid receptor agonist(DADLE)group;Delta opioid receptor antagonist(NTD)group.After 6 h reperfusion,liver tissues histological changes were observed by haematoxylin-eosin(H&E)staining and scored by SUZUKI’s score. Blood serum samples were collected for testing alanine transaminase(ALT)and aspartate transaminase(AST)level. The content of tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) in serum was tested by enzyme linked immunosorbent assay(ELISA). The expression of Janus Kinase(JAK)2 and signal transducer and activators of transcription(STAT)3 mRNA was determined by real-time reverse transcriptionpolymerase chain reaction(RT-PCR). And the JAK and STAT3 proteins were analyzed by Western Blot. Results Compared with the CON group,Delta opioid activation with DADLE significantly reduced serum ALT[(138 ±33.369)U/L vs.(267 ± 106.173)U/L,P < 0.05]and AST[(210 ± 102.666)U/L vs.(374 ± 64.021)U/L,P <0.05]levels. Besides,the SUZUKI′s score of DADLE pretreatment was significantly lower than the CON group(4.83 ± 1.169 vs. 7.67 ± 0.816,P < 0.05). The level of serum TNF-α[(15.561 ± 5.12)pg/mL vs.(32.289 ± 9.23)pg/mL,P < 0.05]and IL-1β[(46.444 ± 7.777)pg/mL vs.(68.841 ± 9.225)pg/mL,P < 0.05]were decreased by DADLE,comparing with the CON group. Moreover,the expression of JAK and STAT3 mRNA was much higher in DADLE group than in the CON group(P < 0.05). The expression of JAK2 and STAT3 protein were also up regulated by DADLE,comparing with the CON group. However,Delta opioid receptor antagonist natrindole reversed these changes. Conclusion Delta opioid receptor activation attenuated hepatic IR injury in rats with cirrhosis,which might associate with up regulating JAK2/STAT3 pathway

关 键 词：Delta阿片受体 缺血再灌注损伤 肝硬化 JAK2/STAT3通路 大鼠 

分 类 号：R73[医药卫生—肿瘤]