重组人白介素-11对脂多糖致新生大鼠空肠上皮细胞系-6损伤的影响

Effects of recombinant human interleukin-11 on LPS-induced intestinal epithelial cell injury in rats

摘要:

目的 探讨重组人白介素-11（rhIL-11）对新生大鼠空肠上皮细胞系-6（IEC-6）增殖和凋亡的影响。方法 通过脂多糖（LPS）作用于正常IEC-6细胞，建立NEC体外模型，为LPS处理组；IL-11治疗组在LPS作用后给予100 ng/mL rhIL-11处理；空白对照组在实验过程中加入等剂量的生理盐水替代。采用MTT比色法选择LPS作用的最佳浓度（5~200 μg/mL）及最佳时间（1~24 h）；MTT 比色法检测rhIL-11 加入后3、6、9和12 h各组细胞的增殖活力；流式细胞术检测各组的细胞凋亡率和坏死率。结果 LPS可降低IEC-6细胞的增殖活力，且根据不同浓度LPS作用于IEC-6细胞不同时间结果，LPS的最佳作用浓度为100 μg/mL，最佳作用时间为3 h。100 ng/mL rhIL-11作用于LPS 处理过的细胞9 h后，IL-11治疗组细胞增殖活力较LPS组显著提高（P<0.05），与空白对照组比较差异无统计学意义（P>0.05）；且IL-11治疗组的总凋亡和坏死率较LPS处理组降低（P<0.01），但仍高于空白对照组（P<0.05）。结论 rhIL-11可促进由LPS致IEC-6细胞损伤后的细胞增殖，有助于IEC-6细胞增殖活力的恢复，降低凋亡和坏死率，提示rhIL-11对LPS致IEC-6细胞损伤具有保护作用。

Abstract:

Objective To study the effects of recombinant human interleukin-11 (rhIL-11) on the proliferation and apoptosis of rat intestinal epithelial cell line (IEC-6). Methods IEC-6 cells were treated with LPS to establish necrotizing enterocolitis (NEC) model in vitro. rhIL-11 (100 ng/mL) was administered following LPS treatment and these cells were used as the IL-11 treatment group. The cells treated with normal saline only served as the control group. MTT assay was used to determine an optimal concentration (5-200 μg/mL) and time (1-24 h). MTT assay was used to measure the proliferation of IEC-6 cells at 3, 6, 9 and 12 hours after rhIL-11 treatment. Flow cytometry was used to evaluate the apoptosis of IEC-6 cells. Results IEC-6 cells treated with various concentrations of LPS at various time points showed a lower proliferation than the control group (P<0.05). After 9 hours of rhIL-11 treatment, the proliferation activity of IEC-6 cells in the IL-11 treatment group significantly increased compared with the NEC model group without rhIL-11 treatment (P<0.05), reaching to the level of the control group. The total apoptotic and necrotic rate of IEC-6 cells in the IL-11 treatment group decreased significantly compared with the NEC model group without rhIL-11 treatment (P<0.01), but were still higher than the control group (P<0.05). Conclusions rhIL-11 can promote proliferation and reduce apoptotic and necrotic rates of IEC-6 cells treated with LPS.