甲状旁腺素对人肾小管上皮细胞凋亡的影响

Effects of parathyroid hormone on apoptosis in cultured renal tubular epithelial cells

目的研究甲状旁腺素(parathyroid hormone,PTH)对体外培养的人肾小管上皮细胞(HK-2)凋亡的影响及其机制。方法不同浓度的PTH刺激HK-2细胞后,应用Hoechst33258染色观察PTH对HK-2细胞形态学的改变,Annexin V/PI双染流式细胞术检测PTH对HK-2细胞凋亡的影响。Western blot和半定量RT-PCR法检测PTH对HK-2细胞Bax和Bcl-2蛋白和基因表达的影响。结果Hoechst33258染色结果表明,PTH作用于HK-2细胞48h,细胞出现核固缩、核碎裂,随着PTH浓度的升高,凋亡细胞数逐渐增多,且具有一定的浓度依赖性。流式细胞术结果表明,随着PTH浓度的升高,凋亡细胞百分率逐渐增多,与对照组比较差异有统计学意义(P<0.05)。Western blot和半定量RT-PCR结果表明,随着PTH浓度的升高和作用时间的延长,Bax蛋白和基因表达量逐渐增加,Bcl-2蛋白和基因表达量逐渐减少,Bax/Bcl-2比值增加。结论PTH可能通过调节凋亡基因Bax和Bcl-2的表达促进肾小管上皮细胞凋亡,进而促进肾小管间质纤维化。

Objective To investigate the effects of parathyroid hormone (PTH) on apoptosis in cultured renal tubular epithelial cells (HK-2) and its mechanism.Methods Cultured HK-2 was treated with PTH of different concentrations. The morphologic changes of HK-2 nuclei were observed by Hoechst 33258 staining,while the apoptosis of HK-2 was detected by flow cytometry via Annexin V/PI staining. Western blot was used to measure the Bax and Bcl-2 protein expressions; RT-PCR was used to measure the Bax and Bcl-2 gene expres...