| 摘要: |
| 目的研究IL-22对ox-LDL诱导的CRL-1730细胞(人脐静脉内皮细胞)凋亡的影响及其机制。 方法100 μg/ml ox-LDL刺激CRL-1730细胞,不同时间点经实时荧光定量RT-PCR检测CRL-1730细胞中IL-22受体R1(IL-22R1)mRNA的表达状况。然后在ox-LDL诱导的CRL-1730细胞凋亡模型中加入20 ng/ml外源性IL-22,通过Annexin Ⅴ-PI凋亡试剂盒检测其对细胞凋亡的影响,经实时荧光定量反转录PCR检测Bcl-2、Bcl-xl、Bcl-w、caspase 3及STAT3 mRNA的表达,采用免疫印迹法分析STAT3磷酸化和Bcl-2蛋白的水平。结果 IL-22R1 mRNA在ox-LDL刺激后表达明显增高(P<0.05),且于12 h达峰值(P<0.01)；加入IL-22可明显抑制ox-LDL刺激诱导的内皮细胞凋亡(P<0.01),促进抗凋亡基因Bcl-2、Bcl-xl和Bcl-w表达,而降低caspase 3 mRNA表达(P<0.01)。此外,IL-22作用1~2 h STAT3发生磷酸化, 4 h后Bcl-2蛋白表达升高(P<0.01)。 结论IL-22可抑制ox-LDL诱导的CRL-1730细胞凋亡,这可能与其促使STAT3磷酸化,上调抗凋亡分子Bcl-2、Bcl-xl和Bcl-w的表达,抑制促凋亡分子caspase 3的表达有关。 |
| 关键词: 白细胞介素 人脐静脉内皮细胞 氧化LDL 细胞凋亡 Bcl-2基因 STAT3蛋白 |
| DOI：10.3724/SP.J.1008.2011.0233 |
| 投稿时间：2010-12-31修订日期：2011-01-20 |
| 基金项目:国家自然科学基金(30770997, 30972730, 81072479), 上海市科委基金(09JC1405400, 08QH14001). |
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| IL-22 inhibits ox-LDL-induced apoptosis and increases Bcl-2 expression in CRL-1730 cells |
| SUN Yi, HU Zhi-de, HUANG Yuan-lan, XIONG Yi-song, DENG An-mei*, ZHONG Ren-qian* |
| (Department of Laboratory Medicine, Changzheng Hospital, Second Military Medical University, Clinical Immunology Center of PLA, ; Key Lab of PLA Clinical Immunology, Shanghai 200003, China) |
| Abstract: |
| ObjectiveTo investigate the effect of interleukin-22 (IL-22) on apoptosis of CRL-1730 cells(human umbilical vein endothelial cell line) induced by ox-LDL and the possible mechanism. MethodsExpression of IL-22 receptor R1 (IL-22R1) mRNA in CRL-1730 cells stimulated with ox-LDL (100 μg/ml) was detected by fluorescence quantitative RT-PCR. Flow cytometry was applied to examine the effect of exogenous IL-22 (20 ng/ml) on the ox-LDL-induced apoptosis of CRL-1730 cells. The expression of Bcl-2, Bcl-xl, Bcl-w, caspase 3, and STAT3 mRNA in CRL-1730 cells treated or untreated with IL-22 and ox-LDL were measured by fluorescence quantitative RT-PCR; the phosphorylation status of STAT3 and the Bcl-2 protein level were examined by Western blotting analysis. ResultsThe expression of IL-22R1 mRNA increased in a time-dependent manner (P<0.05) and peaked at 12 h after ox-LDL stimulation. Treatment with IL-22 significantly inhibited the apoptosis of CRL-1730 cells induced by ox-LDL(P<0.01), increased the expression of anti-apoptosis genes Bcl-2, Bcl-xl, and Bcl-w, and decreased expression of caspase 3 mRNA(P<0.01). STAT3 phosphorylation occurred 1-2 h after IL-22 treatment and Bcl-2 protein expression increased 4 h after IL-22 treatment in ox-LDL stimulated CRL-1730 cells. ConclusionIL-22 can inhibit the pro-apoptosis effect of ox-LDL in the CRL-1730 cells, which might be associated with the increased expression of Bcl-2, Bcl-xl, and Bcl-w mRNA, decreased expression of caspase 3, and induced STAT3 phosphorylation. |
| Key words: IL-22 CRL1730 human umbilical vein endothelial cell ox-LDL apoptosis Bcl-2 Caspase3 STAT3 |
