Rho激酶抑制剂Y-27632对MRL/lpr狼疮小鼠的保护作用

Protection of Rho-associated protein kinase inhibitor Y-27632 in MRL/lpr mice

目的探讨Rho激酶抑制剂Y-27632对MRL/lpr狼疮小鼠的保护作用机制。方法 MRL/lpr狼疮小鼠20只随机分为MRL/lpr对照组、5 mg/kg Y-27632处理组,每组10只;野生型对照组C57BL/6小鼠10只。采用ELISA检测各组小鼠血清、脾脏组织中超氧化物歧化酶(SOD)、丙二醛(MDA)水平,ELISA检测血清核因子κB(NF-κB)相关炎症因子白细胞介素6(IL-6)、IL-1β、肿瘤坏死因子α(TNF-α)含量;采用Western blot法检测各组小鼠脾脏组织中硫氧还蛋白结合蛋白(Txnip)/硫氧还蛋白(Trx)、丝裂原激活蛋白激酶(MAPK)相关蛋白胞外信号调节激酶(ERK)、c-Jun氨基末端激酶(JNK)和p38MAPK以及NF-κB的水平;Western blot法检测各组小鼠脾脏T淋巴细胞Txnip、p38MAPK、NF-κB蛋白水平;ELISA检测T淋巴细胞上清液IL-6、IL-1β、TNF-α水平。结果 Y-27632提高MRL/lpr狼疮小鼠血清及脾脏组织SOD水平;降低血清及脾脏组织MDA水平;降低血清、脾脏组织和脾脏T淋巴细胞上清液IL-6、IL-1β、TNF-α水平;抑制脾脏和脾脏T淋巴细胞Txnip、MAPK相关蛋白ERK、JNK和p38MAPK以及NF-κB表达,增加Trx含量。结论 Rho激酶抑制剂Y-27632对MRL/lpr狼疮小鼠有保护作用。

Objective To investigate the immunological effect of Rho-associated protein kinase inhibitor Y-27632 on MRL/lpr mice and explore its underlying mechanisms. Methods The study collected 10 wild-type C57BL/6 mice as normal controls and 20 MRL/lpr mice that were randomly assigned to MRL/lpr group and 5 mg/kg Y-27632-treated MRL/lpr group,with 10 mice in each group. Superoxide dismutase（ SOD）,methane dicarboxylic aldehyde（ MDA）,interleukin 6（ IL-6）,IL-1β and tumor necrosis factor α（ TNF-α） in the serum or spleen were detected by ELISA. The protein levels of thioredoxininteracting protein/thioredoxin（ Txnip/Trx）,MAPK-associated protein ERK,JNK,p38 and NF-κB were detected by Western blot analysis. T lymphocytes were isolated and cultured from the groups,and then subjected to Western blotting for evaluating the levels of Txnip/Trx,MAPK,NF-κB proteins and to ELISA for determining the levels of IL-6,IL-1β,TNF-α in the supernatants.Results Compared with the MRL/lpr group,the level of SOD increased and MDA decreased in the serum and spleen; the levels of IL-6,IL-1β,TNF-α were reduced in the serum,spleen and the supernatant of T cells from the spleen; the protein levels of Txnip/Trx,MAPK,NF-κB from the spleen were inhibited and the level of Trx in the spleen was raised in MRL/lpr mice treated with Y-27632. Conclusion Ros kinase inhibitor Y-27632 has protective effects on MRL/lpr lupus mice.