不同剂量γ射线对肾小管上皮细胞增殖与功能的影响

Effects of Different Dose of Gamma Rays on Renal Tubule Epithelial Cells: Proliferation and Function.

目的研究受不同剂量γ射线照射后,原代培养肾小管上皮细胞增殖和功能的改变情况。方法机械过滤加酶消化法分离培养肾小管上皮细胞,以01、、51、0 Gyγ射线照射细胞,MTT法测定细胞的增殖,RT-PCR法分析基因mRNA表达,Western Blot法检测相关蛋白的表达。结果肾小管上皮细胞受1 Gyγ射线照射后对数生长期延迟,受5 Gy1、0 Gyγ射线照射后不出现对数生长期。受1 Gy、5 Gy1、0 Gyγ射线照射的肾小管上皮细胞Na+,K+-ATP酶、1α羟化酶、24羟化酶mRNA表达明显降低,TGF-βmRNA和蛋白质表达明显升高。受1Gy射线照射后OPN蛋白量提高,而受5 Gy、10 Gy照射后OPN蛋白量降低。结论不同剂量γ射线可明显抑制肾小管上皮细胞的增殖,促使肾小管上皮细胞向成纤维细胞转化,并通过增加TGF-β的分泌促进纤维蛋白等基质蛋白在细胞外的堆积,这种作用存在明显的剂量依赖关系。

Objective To study changes of proliferation and function of primary renal cells after being exposexl to different dose of gamma rays. Methods Primary renal cells were cultivated by means of mechanical separation and enzyme digestion. Cells were subjected to 0, 1, 5, and 10 Gy gamma rays, and proliferation of cells was detected by MTT, expression of mRNAs determined by RT - PCT, expression of proteins by Western Blot. Results Logarithmic growth phase of cells was delayed by 1 Gy gamma rays, and restrained by 5Gy and 10 Gy gamma rays. The expression of mRNA of Na^＋ , K^＋ - ATP enzyme, 1α - hydroxylase and 24 - hydroxylase was significantly decreased by 1Gy, 5Gy and 10Gy gamma rays. The expression of mRNA and protein of TGF - βwas significantly promoted by 1Gy, 5Gy and 10Gy gamma rays. The expression of protein of OPN was promoted by 1 Gy gamma rays, and inhibited by 5Gy and 10Gy gamma rays. Conclusion Irradiatied renal cells with different dose of gamma rays can significantly suppress its proliferation, promoting it turn into fibroblast, and resulting in accumulation of matrix proteins by way of increasing TGF -β secretion. This effect is dose-dependent.