类胰蛋白酶通过激活PAR-2促进肠黏膜上皮IEC-6细胞损伤  

Role of PAR-2 in tryptase mediated IEC-6 cell injury

作　　者：李顺[1] 黄品婕[1] 葛缅[1] 甘小亮[2] 

机构地区：[1]中山大学附属第三医院麻醉科,广东广州510630 [2]中山大学中山眼科中心麻醉科,广东广州510060

出　　处：《中国病理生理杂志》2015年第3期530-533,共4页Chinese Journal of Pathophysiology

基　　金：广州市科技计划(No.2014J4100172);中山大学青年教师培育项目(No.12ykpy37)

摘　　要：目的:观察类胰蛋白酶(tryptase)对小肠黏膜上皮细胞IEC-6的作用及机制。方法:体外培养IEC-6细胞,随机分为对照(control)组、tryptase处理组、蛋白酶激活受体2(PAR-2)抑制剂(FSLLRY-NH2,FS)处理组以及tryptase+FS处理组。采用MTT法检测细胞生存率;测定分泌的乳酸脱氢酶(LDH)活性;Western blotting测定PAR-2和cleaved-caspase 3的表达。结果:与对照组比,100μg/L和1 000μg/L类胰蛋白酶导致细胞生存率明显降低(P<0.05)。与对照组相比,10μg/L到1 000μg/L类胰蛋白酶导致LDH的活性明显增加(P<0.05)。与对照组相比,100μg/L和1 000μg/L类胰蛋白酶导致细胞的PAR-2及cleaved-caspase 3表达明显升高(P<0.05)。与1 000μg/L tryptase处理组比较,FS能够明显增加细胞生存率、降低LDH活性及cleaved-caspase 3表达(P<0.05)。结论:类胰蛋白酶通过激活PAR-2促进IEC-6细胞损伤,且这种损伤呈浓度依赖性。AIM：To investigate the role of protease activated receptor-2 （ PAR-2 ） in the process of tryptase mediated IEC-6 cell injury.METHODS：The rat intestinal epithelial cell line IEC-6 was treated with tryptase at different concentrations （1 μg/L, 10 μg/L, 100μg/L and 1 000μg/L） in the presence or absence of PAR-2 antagonist FSLLRY-NH2 for 12 h respectively.The cell survival rate was detected by MTT assay.The protein levels of PAR-2 and cleaved-caspase 3 were determined by Western blotting.The LDH activity was also measured.RESULTS：Compared with control group, the cell survival rates were significantly decreased in 100 μg/L and 1 000 μg/L tryptase treated groups, the LDH activities were significantly increased in 10 μg/L to 1 000 μg/L tryptase treated groups, and the protein levels of PAR-2 and cleaved caspase 3 were significantly increased in 100μg/L and 1 000μg/L tryptase treated groups （P〈0.05）.Com-pared with 1 000 μg/L tryptase treated group, the LDH activity and cleaved caspase 3 protein level were dramatically de-creased while the survival rate was significantly increased in the presence of PAR-2 antagonist FSLLRY-NH2 （P〈0.05）. CONCLUSION：Tryptase induces IEC-6 cell injury in a dose-dependent manner by activating PAR-2.

关 键 词：类胰蛋白酶 蛋白酶激活受体2 IEC-6细胞 乳酸脱氢酶 Caspase 3 

分 类 号：R363.2[医药卫生—病理学]