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  

Role of parathyroid hormone-related peptide in transdifferentiation of NRK-52E cells induced by high glucose

作　　者：梁华晟 钟宇华 

机构地区：[1]广西北海市内分泌代谢病研究所,广西北海536000

出　　处：《第三军医大学学报》2010年第11期1165-1168,共4页Journal of Third Military Medical University

基　　金：广西科技厅自然科学基金(0991294);广西北海市科学研究与技术开发计划项目(200901059)~~

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5mmol/L葡萄糖DMEM培养基培养)组及高浓度葡萄糖组(含16.7mmol/L葡萄糖DMEM培养基培养)中甲状旁腺素相关肽(parathyroid hormone-related peptide,PTHrP)及其受体1在大鼠肾小管导管上皮细胞株NRK-52E表达。10pmol/L PTHrP及16.7mmol/L葡萄糖分别干预72h,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(Type1receptor 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。Objective To study the role of parathyroid hormone-related peptide （ PTHrP） in transdif-ferentiation of NRK-52E cells induced by high glucose. Methods Expression of PTHrP and its receptor 1 in NRK-52E cells incubated with 0 （ control group） ,5 （ normal glucose group） and 16. 7mmol/L glucose （ high glucose group） ,respectively,was detected by RT-PCR and Western blotting,respectively. Expression of TGF-β1,MMP2,type Ⅰ collagen and α-SMA in NRK-52E cells of normal and high glucose groups was detected by Western blotting and ROS was detected by CM2H2DCFDA after 72 h. Results The expression levels of PTHrP,PTHrP receptor 1 （ PTH1R） ,and PTHrP mRNA were higher in high glucose group than in control and normal glucose groups （ P 0. 05）. The expression of TGF-β1,type Ⅰ collagen,and α-SMA after interfered with PTHrP was significantly higher in normal and high glucose groups than in control group （ P 0. 05）. The expression of MMP2 was up-regulated after interference with PTHrP and high glucose level （ P 0. 01）. The PTHrP and high glucose level increased the ROS in NRK-52E cells （ P 0. 01）. Conclusion High glucose level can influence the transdifferentiation of NRK-52E cells by regulating PTHrP and its receptor level.

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 

分 类 号：R322.61[医药卫生—人体解剖和组织胚胎学] R394.1[医药卫生—基础医学]