坏死细胞释放IL-1对血管平滑肌细胞增殖及炎性反应的影响

目的 研究坏死血管平滑肌细胞对周围正常细胞增殖及炎性因子分泌的影响,并探讨其可能的作用机制.方法 无血清低糖低氧条件下培养细胞,建立细胞坏死模型,收集坏死细胞培养上清液,用于干预正常血管平滑肌细胞.实验设坏死上清组、IL-1组、坏死上清+IL-1RA组和对照组.MTT法检测各组细胞增殖情况；RT-PCR检测各组MCP-1、IL-6和IL-8 mRNA的表达；ELISA检测各组MCP-1、IL-6和IL-8的分泌；Western blot检测各组NF-kB的激活情况.结果 NCS组和IL-1组细胞增殖能力均显著高于对照组(P＜0.01),NCS+IL-1 β组细胞增殖能力显著低于NCS组和IL-1组(P＜0.01)；NCS组和IL-1 β组MCP-1、IL-6、IL-8 mRNA的表达均显著高于对照组(P＜0.05),NCS+IL-1RA组MCP-1、IL-6、IL-8 mRNA的表达均显著低于NCS组和IL-1组(P＜0.05)；NCS组和IL-1 β组的MCP-1、IL-6及IL-8蛋白的分泌均显著高于对照组(P＜0.05),NCS+IL-1RA组MCP-1、IL-6和IL-8蛋白的分泌显著低于NCS组及IL-1组(P＜0.05)；NCS组和IL-1组NF-kB的活性显著高于对照组(P＜0.05),与NCS组和IL-1组相比NCS+IL-1RA组NF-kB的活性显著降低(P＜0.05).结论 坏死血管平滑肌细胞能够促进正常细胞的增殖,诱导NF-kB的激活以及炎性因子的释放,并且这种促进作用可能与IL-1的大量释放有关.

Influence of IL-1 Released by Necrotic Cells on Vascular Smooth Muscle Cell Proliferation and Inflammatory Reaction

Objective To study the the necrotic vascular smooth muscle cells, and to explore its possible mechanism of action of the surrounding normal cell proliferation and inflammatory cytokine secretion. Methods Cells were cultured in low oxygen serum free and glucose deprivation medium to establish cell necrosis model. Necrotic cell culture supernatant was collected to interfere normal vascular smooth muscle cell. Experiment groups were set as necrosis supematant group, IL-1 group, necrosis supernatant of IL-1RA group and control group, MTI＂ assay of cell proliferation; RT-PCR was used to detect the groups of MCP-1, IL-6, IL-8 mRNA expression; ELISA to detect the MCP-1, IL-6, IL-8 secretion; Western blot to detect the activation of NF-kB. Results NCS group and IL-1 cell proliferation were significantly higher than that in the control group （P〈0.01）, NCS Group IL-1 beta cell proliferation was significantly lower than that of the NCS group and IL-1 group （P〈0.01）; NCS group and IL-1 3, MCP-1, II~6, IL-8 mRNA expression were significantly higher than those in the control group （P〈0.05） the NCS of IL- IRA groups MCP-1, IL-6, IL-8 mRNA expression was significantly lower than the NCS group and IL-1 group （P〈0.05）; NCS group and IL-1 β MCP-1, IL-6, IL-8 protein secretion, significantly higher than the control group （P〈0.05）, the NCS of IL-1RA groups MCP-1, IL-6, IL-8 protein secretion was significantly lower than the NCS group and IL-1 group （P〈0.05）, the NCS group and IL -1 NF-kB activity was significantly higher than that of the control group （P〈0.05）, compared with the NCS group and IL- 1 group NCS IL-1RA group of NF-kB activity was significantly decreased （P〈0.05）. Conclusion The necrosis vascular smooth muscle cells can promote the proliferation of normal cells, induction of NF-kB activation and inflammatory cytokine release, and this role may be related to the massive release of IL-1.