过表达NHE1通过上调钙蛋白酶活性降低RAW264.7细胞ABCA1蛋白表达

发布时间：2018-04-28 08:56

本文选题：钠氢交换体(NHE) + 钙蛋白酶(calpain)　；参考：《细胞与分子免疫学杂志》2017年01期

【摘要】：目的探讨过表达钠氢交换体1(NHE1)对RAW264.7细胞三磷酸腺苷结合盒转运体A1(ABCA1)蛋白表达的影响。方法采用Ad NHE1腺病毒感染RAW264.7细胞,Western blot法检测NHE1-EGFP融合蛋白的表达,激光共聚焦显微镜技术检测NHE1-EGFP融合蛋白细胞内定位,酸负载p H回复检测NHE1活性,Western blot法检测NHE1-EGFP融合蛋白对RAW264.7细胞ABCA1蛋白水平及钙蛋白酶(calpain)活性的影响,加入calpain抑制剂N-乙酰基-L-亮氨酰-L-亮氨酰-L-正亮氨酸(ALLN),Western blot法检测肝脏X受体激动剂TO-901317诱导ABCA1蛋白表达水平的影响。结果 Ad NHE1腺病毒感染RAW264.7细胞后,高表达NHE1-EGFP融合蛋白,定位在细胞质及细胞膜。NHE1-EGFP融合蛋白可降低ABCA1蛋白的表达水平并提高calpain活性,而calpain抑制剂ALLN能阻断ABCA1蛋白表达水平降低。结论 NHE1过表达通过上调calpain活性而降低ABCA1蛋白表达水平。
[Abstract]:Aim to investigate the effect of overexpression of sodium hydrogen exchanger 1 (NHE1) on the expression of adenosine triphosphate binding cassette transporter A1 (A1) ABCA1 protein in RAW264.7 cells. Methods Ad NHE1 adenovirus infected RAW264.7 cells were used to detect the expression of NHE1-EGFP fusion protein by Western blot, and laser confocal microscopy was used to detect the intracellular localization of NHE1-EGFP fusion protein. The effect of NHE1-EGFP fusion protein on the level of ABCA1 protein and calpain activity in RAW264.7 cells was detected by Western blot assay. The effect of calpain inhibitor, N-acetyl-L-leucyl-L-leucine-L-leucine, on the expression of ABCA1 protein induced by TO-901317, a liver X receptor agonist, was detected by Western blot. Results after Ad NHE1 adenovirus infected RAW264.7 cells, the high expression of NHE1-EGFP fusion protein was found, which was located in cytoplasm and cell membrane. NHE1-EGFP fusion protein could decrease the expression level of ABCA1 protein and increase calpain activity, while calpain inhibitor ALLN could block the decrease of ABCA1 protein expression level. Conclusion overexpression of NHE1 can decrease the expression of ABCA1 protein by up-regulating the activity of calpain.
【作者单位】：贵州医科大学附属医院综合病房;贵州医科大学贵州省医学分子生物学重点实验室;贵州医科大学免疫学教研室;
【基金】：国家自然科学基金(31260250)
【分类号】：R363

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