贝前列素钠对急性重症胰腺炎大鼠c-JNK信号通路的影响

Effect of beraprost sodium on c-JNK signal pathway in rats with severe acute pancreatitis

目的:观察贝前列素钠(beraprost sodium)对急性重症胰腺炎(severe acute pancreatitis,SAP)大鼠c-Jun氨基末端激酶(c-Jun N-terminalkinase,c-JNK)信号通路的影响.方法:48只♂SD大鼠,随机分为假手术组(正常对照组,NC组)、SAP组(S组)和贝前列腺素钠组(BS组).S组和BS组大鼠腹腔注射盐酸氯胺酮麻醉下逆行胰胆管内加压注射5%牛磺胆酸钠建立SAP模型.S组大鼠给予腹腔注射生理盐水,BS组大鼠腹腔注射贝前列素钠20μg/kg干预组.最终纳入实验各组分别为13、12及11只.在实验进行的第1、3、6、12 h时间点分别处死相应大鼠并采集胰腺标本,运用酶联免疫吸附实验(ELISA)法测定胰腺组织的肿瘤坏死因子α(tumor necrosisf a c t o rα,T N F-α)、白介素-6(i n t e r l e u k i n-6,IL-6)的含量;反转录聚合酶链式反应(reversetranscription-polymerase chain reaction,RTP C R)法检测胰腺组织中c-J N K m R N A的变化;免疫印迹(Western blot)法检测各组大鼠胰腺组织中c-JNK蛋白及磷酸化c-JNK(p-cJ N K)蛋白活性.采用定量资料3×4析因设计的方差分析,运用SPSS17.0统计软件包进行统计分析.结果:与NC组比较,S组胰腺组织TNF-α(458.7p g/m L±36.9 p g/m L v s 197.1 p g/m L±5.3p g/m L)、I L-6(525.6 p g/m L±12.6 p g/m Lv s 2 11.4 p g/m L±3.5 p g/m L)、c-J N Km R N A(2.60±0.09 v s 0.91±0.59)水平增高;c-JNK蛋白(1.52±0.41 vs 0.98±0.11)及p-cJ N K蛋白(1.53±0.45 v s 0.98±0.08)活性增强.与S组相比,BS组TNF-α(211.7 pg/m L±3.4 pg/m L vs 458.7 pg/m L±36.9 pg/m L)、I L-6(468.7 p g/m L±34.9 p g/m L v s 525.6pg/m L±12.6 pg/m L)、c-JNK m RNA(1.93±0.14 vs 2.60±0.09)、c-JNK蛋白(1.32±0.35vs 1.52±0.41)及p-c-JNK蛋白(1.22±0.37 vs 1.53±0.45)水平下降,差异均有统计学意义(P<0.05).结论:c-JNK通路在SAP大鼠胰腺组织损伤中起一定作用;贝前列腺素钠治疗SAP可能是通过c-JNK通路实现的,为贝前列腺素钠用于治疗SAP提供一定的理论依据.

AIM： To investigate the effect of beraprost sodium （BS） on the c-Jun N-terminal kinase （c-JNK） signal pathway in rats with severe acute pancreatitis （SAP）. METHODS： Forty-eight SD rats were randomly divided into a sham operation group （normal control）, an SAP group and a BS group. SAP was induced by retrograde injection of 5% sodium tauroeholate. BS was given at a dose of 20 μg/kg at the time of pancreatitis induction in the BS group. At 1, 3, 6, and 12 h after SAP was successfully induced, the animals were killed. Levels of tumor necrosis factor α （TNF-α） and interleukin-6 （IL-6） in pancreatic tissue were detected by ELISA, c-JNK mRNA expression was detected by RT-PCR, and c-JNK and phosporylated c-JNK （p-c-JNK） protein in pancreatic tissue was examined by Western blot. RESULTS： Compared with the normal control group, the levels of TNF-α, IL-6, c-JNK mRNA, c-JNK protein and p-c-JNK protein in pancreatic tissues were significantly enhanced in the SAP group （TNF-α： 458.7 pg/mL ± 36.9 pg/mL vs 197.1 pg/mL ± 5.3 pg/mL; IL-6： 525.6 pg/mL ± 12.6 pg/mL vs 211.4 pg/mL ± 3.5 pg/mL; c-JNK mRNA： 2.60 ± 0.09 vs 0.91 ± 0.59; c-JNK protein： 1.52 ± 0.41 vs 0.98 ± 0.11; p-c-JNK protein： 1.53 ± 0.45 vs 0.98 ± 0.08, P 〈 0.05 for all）. Compared with the SAP group, the levels of TNF-α, IL-6, c-JNK mRNA, c-JNK protein and p-c-JNK protein were significantly decreased in the BS group （TNF-α： 211.7 pg/mL ± 3.4 pg/mL vs 458.7 pg/m L ± 36.9 pg/mL; IL-6： 468.7 pg/mL ± 34.9 pg/mL vs 525.6 pg/mL± 12.6 pg/mL; c-JNK mRNA： 1.93 ± 0.14 vs 2.60 ± 0.09; c-JNK protein： 1.32 ± 0.35 vs 1.52 ± 0.41; p-c-JNK protein： 1.22 ± 0.37 vs 1.53 ± 0.45, P 〈 0.05 for all）. CONCLUSION： BS can alleviate the inflammatory response in SAP rats by down-regulating c-JNK signaling.