美沙拉嗪通过调节钙粘蛋白11对克罗恩大鼠肠上皮细胞凋亡和炎症反应的影响

Effect of mesalazine on apoptosis and inflammatory response of intestinal epithelial cells in crohn rats through regulation of calmodulin 11

目的探究美沙拉嗪对克罗恩大鼠肠上皮细胞凋亡和炎症反应的影响及机制。方法用2,4,6-三硝基苯磺酸(TNBS)3 mL·kg^(-1)灌肠的方法构建大鼠克罗恩病模型。将30只雄性SD大鼠分为对照组、模型组和实验组,每组10只。模型组和实验组大鼠构建克罗恩病模型,对照组大鼠给予0.9%氯化钠3 mL·kg^(-1)灌肠。随后模型组和对照组大鼠灌胃0.9%氯化钠溶液2 mL·kg^(-1),实验组大鼠灌胃美沙拉嗪2 mL·kg^(-1)。(1)组织实验:用酶联免疫吸附法(ELISA)测定大鼠结肠组织白细胞介素8(IL-8)、IL-1β、IL-4和IL^(-1)0的含量;用蛋白免疫印迹法检测大鼠结肠组织内钙粘蛋白11(CDH11)的表达水平。(2)细胞实验:用CCK-8实验检测各组细胞存活率;用Transwell法检测各组细胞侵袭能力;用原位末端标记检测法检测各组细胞凋亡率。结果(1)组织实验结果:对照组、模型组和实验组大鼠结肠组织内IL-8含量分别为(378.21±2.64),(614.55±2.17)和(416.35±2.24)pg·mL^(-1);这3组CDH11蛋白的表达水平分别为1.00±0.41,2.53±1.25,1.67±0.12。(2)细胞实验结果:对照组、模型组和实验组大鼠结肠上皮细胞在72 h细胞存活率分别为(100.00±0.00)%,(57.45±4.06)%和(64.65±6.07)%;这3组细胞侵袭数目分别为(85.45±9.47),(100.45±9.35)和(91.12±8.47)个;这3组细胞凋亡率分别为(9.12±1.13)%,(27.71±2.53)%和(15.57±1.86)%。上述指标,对照组与模型组比较,差异均有统计学意义(均P<0.01);模型组和实验组比较,差异均有统计学意义(均P<0.01)。结论美沙拉嗪可通过抑制CDH11的表达,改善TNBS诱导的克罗恩大鼠炎症反应和肠上皮细胞损伤。

Objective To investigate the effect and mechanism of mesalazine on apoptosis and inflammatory response of intestinal epithelial cells in Crohn rats.Methods A rat Crohn’s disease model was constructed by using 2,4,6-trinitrobenzenesulfonic acid (TNBS) 3m L·kg;enema.Thirty male SD rats were randomly divided into control group,model group and test group,10 rats in each group,in which the model and test group rats were constructed as Crohn disease models,and the control group rats were given 0.9%sodium chloride 3m L·kg;enema.Subsequently,rats in the model and control groups were gavaged with 0.9%NaCl solution 2 m L·kg;,and rats in the test group were gavaged with mesalazine 2 mL·kg;.(1) Tissue experiment:rat colon tissue interleukin 8 (IL-8),IL-1β,IL-4,IL-10 content were measured by enzymelinked immunosorbent assay;The expression level of calmodulin 11 (CDH11) protein in rat colon tissue was detected by protein immunoblotting (Western blot).(2) Cellular experiment:The cell survival rate of each group was detected by CCK-8 assay;the invasion of each group was detected by Transwell assay;the apoptosis rate of each group was detected by in situ end-labeling assay.Results (1) Results of tissue experiments:The levels of IL-8 in the colon tissue of rats in the control group,model group and test group were (378.21±2.64),(614.55±2.17),(416.35±2.24) pg·m L;,respectively;the expression level of CDH11 protein were 1.00±0.41,2.53±1.25,1.67±0.12,respectively.(2) Cellular experiment results:the cell survival rates of colonic epithelial cells in the control group,model group and test group at the 72 h were (100.00±0.00)%,(57.45±4.06)%,(64.65±6.07)%;the number of cell invasion were (85.45±9.47),(100.45±9.35),(91.12±8.47);the apoptosis rate were(9.12±1.13)%,(27.71±2.53)%,(15.57±1.86)%.For the above indicators,comparison between control group and model group,the difference was significantly (P<0.01);Comparison between model group and test group,the difference was significantly (P<0.01).Conclusion Mesalazine ameliorates TNBS-induced inflammatory response and intestinal epithelial cell injury in Crohn’s rats by inhibiting the expression of CDH11.