甲状旁腺激素相关肽在高糖诱导NRK-52E细胞转分化中的作用.docVIP

甲状旁腺激素相关肽在高糖诱导NRK-52E细胞转分化中的作用[ [基金项目] 广西科技厅自然科学基金资助项目（0991294）；广西北海市科学研究与技术开发计划项目（200901059） [通信作者] 梁华晟，电话: （0779） 2022492, E-Mail:flowchaos@ 梁华晟，钟宇华 （536000 广西 北海，广西北海市内分泌代谢病研究所）（广西医科大学第九附属医院内分泌科日前已经正式更名为广西北海市内分泌代谢病研究所） [摘要] 目的 探讨甲状旁腺素相关肽在高糖诱导大鼠肾小管导管上皮细胞株NRK-52E转分化作用。方法 应用RT-PCR及Western blot检测无糖组（无糖培养基培养），正常葡萄糖组（含5 mmol/ L葡萄糖DMEM培养基培养）组及高浓度葡萄糖组（含16.7 mmol/L葡萄糖DMEM培养基培养）中甲状旁腺素相关肽（parathyroid hormone-related protein ，PTHrP）及其受体1在大鼠肾小管导管上皮细胞株NRK-52E表达。10 pmol/L PTHrP及16.7 mmol/L葡萄糖分别干预72 h，应用Western blot检测转化因子β、金属蛋白酶2、Ⅰ型胶原及α平滑肌肌动蛋白表达情况。ROS含量应用CM2H2DCFDA试剂盒检测。结果 与空白组和正常葡萄糖组相比，高浓度葡萄糖能显著上调PTHrP(P＜0.05, P＜0.01)和甲状旁腺激素受体1(Type 1 receptor parathyroid hormone-related protein，PTH1R) (P＜0.01, P＜0.05)蛋白及 PTHrP mRNA(P＜0.01)表达，PTH1R mRNA在高浓度及正常葡萄糖浓度组的表达高于空白组(P＜0.01)。对比空白组，PTHrP能显著上调NRK-52E细胞转化因子β1(P＜0.01)、Ⅰ型胶原(P＜0.01)及α平滑肌肌动蛋白(P＜0.01)表达，而在高浓度葡萄糖状态下，其表达上调更显著(P＜0.05, P＜0.01)。金属蛋白酶2在PTHrP及高浓度葡萄糖共同干预下表达上调(P＜0.01)。PTHrP及高血糖均可以升高NRK-53E细胞ROS含量(P＜0.01, P＜0.01)。结论 高浓度葡萄糖可能通过调控PTHrP/PTH1R的水平从而影响肾小管导管上皮转分化。 [关键词] 甲状旁腺激素相关蛋白质；细胞转分化；NRK-52E细胞；葡萄糖；体外研究 [中图法分类号] [文献标志码] A Effect of PTHrP on hyperglycemia induced transdifferentiation in NRK-52E Liang Huasheng, Zhong Yuhua （Beihai Institute of Endocrine and Metabolic Diseases, Beihai, Guangxi, 536000, China) [Abstract] Objective To study whether PTHrP and PTH1R had affected hyperglycemia to induce transdifferentiation in NRK-52E. Methods PTHrP and PTH1R were detected by RT-PCR and Western blot in control group(0 mmol/L glucose) , normal glucose group(5 mmol/L glucose), high glucose group(16.7mmol/L glucose). Then the expression of TGF-β1, MMP2, type Ⅰ collagen and α-SMA in NRK-52E cells treated by 10pmol/L and/or 16.7mmol/L glucose were detected by Western blot and ROS were detected by CM2H2DCFDA after 72h Result Contrast with control and normal glucose group, the expression of PTHrP (P＜0.05, P＜0.01)，PTH1R (P＜0.01, P＜0.05) and PTHrP mRNA (P＜0.01, P＜0.01) were up-regulated by hyperglycemia, which of PTH1R mRNA in control group