摘要
目的探讨去脂牛血清白蛋白(d-BSA)超载大鼠肾小管上皮细胞(NRK-52E)对内质网应激(ERS)经典标志物葡萄糖调节蛋白78(GRP78)、氧调节蛋白150(ORP150)及细胞凋亡率的影响。方法以不同浓度(1、5、10、20、50mg/ml)的d-BSA超载NRK-52E细胞为实验组,以正常培养的NRK-52E细胞为对照组,观察各组细胞GRP78、ORP150和细胞凋亡率的情况;观察作用时间(6、12、24h)对d-BSA(20mg/ml)超载的NRK-52E细胞GRP78、ORP150和细胞凋亡率的影响情况。通过免疫细胞化学法测定NRK-52E细胞GRP78的表达量,Westernblot法测定ORP150的表达量,流式细胞术检测NRK-52E细胞的凋亡率情况。结果 (1)浓度为1mg/ml的d-BSA孵育NRK-52E细胞24h后GRP78、ORP150的表达量及细胞凋亡率与对照组相比差异无统计学意义(P>0.05)。(2)浓度为5mg/ml的d-BSA孵育NRK-52E细胞24h后,细胞内GRP78、ORP150表达量与对照组相比差异有统计学意义(P<0.05),且浓度为10、20、50mg/ml的d-BSA组的表达量显著增加(P<0.01)。(3)浓度为5mg/ml的d-BSA孵育NRK-52E细胞12h后,细胞凋亡率升高,与对照组相比差异有统计学意义(P<0.05),且浓度为10、20、50mg/ml的d-BSA组细胞凋亡率显著增加(P<0.01)。(4)浓度为20mg/ml的d-BSA孵育NRK-52E细胞6h后,细胞内GRP78、ORP150表达量即有增加(P<0.05);12、24h后,两者表达量与对照组相比,差异有统计学意义(P<0.01)。(5)细胞凋亡率在浓度20mg/ml的d-BSA孵育6h后即升高,但与对照组相比无统计学差异;12、24h后,细胞凋亡率与对照组相比差异有统计学意义(P<0.01)。结论 d-BSA超载NRK-52E细胞可以促进细胞内GRP78、ORP150的表达;当d-BSA浓度过大或者孵育时间过久,细胞凋亡率升高。
Objective To investigate the effects of d-BSA-induced on GRP78, ORP150 and apoptosis of NRK-52E cells. Methods NRK-52E cells were induced by different concentrations of d-BSA （1, 5, 10,20,50 mg/ ml）, the control group was the normal rat kidney proximal tubular epithelial ceil line（NRK-52E）. The expressions of GRP78, ORP150 and the apoptosis rate of the NRK-52E cells of all the groups were observed. The induced time （6, 12,24 h）whether influenced on expressions of GRP78, ORP150 and the apoptosis rate of the NRK-52E cells or not were observed when induced by d-BSA（20 mg/ml）. GRP78 was determined by immunocytochemistry and ORP150 by Western blot. The apoptosis was detected by flow cytometry （ FCM ）. Results （ 1 ） After NRK-52E cells were induced by 1 mg/ml d-BSA after 24 h, the expressions of GRP78, ORPIS0 and apoptosis had no statistical significance compared with normal control group（P 〉 0. 05 ）. （2） The expressions of GRP78 and ORP150 of NRK- 52E cells were upregulated after induced by 5 mg/ml d-BSA-induced for 6 h compared with the normal control group （P 〈0. 05） ;10,20,50 mg/nd d-BSA groups were upregulated significantly compared with the normal control group （P 〈 0. 01 ）. （3）The apoptosis rate of NRK-52E cells was increased by 5 mg/ml d-BSA-induced for 12 h compared with normal control group （ P 〈 0. 05 ） ; 10,20,50 mg/ml d-BSA groups increased significantly （ P 〈 0. 01 ）. （4） After NRK-52E cells were induced by 20 mg/ml d-BSA for 6 h,the expressions of GRP78 and ORP150 were increased compared with control group （P 〈 0. 05） ;for 12,24 h, the expressions of GRP78 and ORP150 were increased significantly compared with control group（ P 〈 0. 01 ）. （5）After NRK-52E cells were induced by 20 mg/ml d-BSA for 6 h,the apoptosis rate was increased compared with the normal control group, but had no significant difference ;for 12 and 24 h, there were significant difference（P 〈 0. 01 ）. Conclusions D-BSA may increase the expressions of GRP78 and ORP150 of NRK-52E cells;ERS may subsequently lead to apoptosis.
出处
《中华临床医师杂志（电子版）》
CAS
2012年第10期106-109,共4页
Chinese Journal of Clinicians(Electronic Edition)
基金
江苏省重点医学人才基金(RC2007115)
