PC12细胞铝染毒后代谢型谷氨酸受体1在PKC和NMDAR表达中的调控作用  

Regulation of mGluR1 on expression of PKC and NMDAR in PC12 exposed to aluminum maltolate

作　　者：王翡 张婷 李立荣 薛星莉 牛侨 WANG Fei;ZHANG Ting;LI Li-rong;XUE Xing-li;NIU Qiao(School of Public Health,Shanxi Medical University,Taiyuan,Shanxi 030001,China)

机构地区：[1]山西医科大学公共卫生学院劳动卫生学教研室

出　　处：《环境与职业医学》2018年第11期1025-1030,共6页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金重点项目(编号:81430078)

摘　　要：[目的]探讨激动和抑制代谢型谷氨酸受体1(mGluR1)蛋白表达对麦芽酚铝染毒的大鼠肾上腺髓质嗜铬瘤分化细胞株(PC12)蛋白激酶C(PKC)及N-甲基-D-天冬氨酸受体(NMDAR)表达的影响。[方法]取对数生长期PC12细胞,分为空白对照组(正常PC12细胞)、激动剂组(100μmol/L mGluR1激动剂)、抑制剂组(100μmol/L mGluR1抑制剂)、麦芽酚铝染毒组(200μmol/L麦芽酚铝)、染铝+激动剂组(100μmol/L mGluR1激动剂+200μmol/L麦芽酚铝)、染铝+抑制剂组(100μmol/L mGluR1抑制剂+200μmol/L麦芽酚铝),染毒时间为24 h。采用实时荧光定量PCR法测定各组PC12细胞中PKC及NMDAR亚基的mR NA表达水平;采用Western blot法测定各组PC12细胞中PKC及NMDAR亚基的蛋白表达水平;采用ELISA法测定各组PC12细胞的PKC酶活性。[结果]与空白对照组相比,麦芽酚铝染毒组的PKC、NMDAR1、NMDAR2A和NMDAR2B mRNA表达水平分别下降39%、21%、38%和26%,差异有统计学意义(P <0.05),其蛋白表达水平分别下降39%、31%、41%和46%,差异有统计学意义(P <0.05)。染铝+抑制剂组的NMDAR1 mRNA和蛋白表达与麦芽酚铝染毒组相比分别上调32%和36%(P <0.05);与麦芽酚铝染毒组相比,染铝+抑制剂组NMDAR2B mRNA表达下调46%,染铝+激动剂组NMDAR2B蛋白表达上调95%,差异有统计学意义(P <0.05)。与对照组相比,麦芽酚铝染毒组PKC酶活性下降56%,而染铝+激动剂组PKC酶活性相对于麦芽酚铝染毒组上调116%(P <0.05)。[结论]麦芽酚铝可以抑制PC12细胞的PKC和NMDAR各亚基的mRNA和蛋白表达;麦芽酚铝可通过mGluR1调节NMDAR表达和PKC酶活性;mGluR1对NMDAR的调节主要作用于NMDAR1和NMDAR2B。[Objective]To investigate the effects of agitating and inhibiting mGluR1on protein kinase C (PKC)and N-methyl-D-aspartate receptor (NMDAR)expressions in rat adrenal-derived pheoehromoeytoma cells (PC12)induced by aluminum maltolate. [Methods ]PC12 cells at logarithmic growth phase were divided into a control group (normal PC12),an agonist group (100μmol/L mGluR1agonist),an inhibitor group (100μmol/L mGluR1 inhibitor),an aluminum maltolate exposure group (200μmol/L aluminum maholate),an aluminum maltolate +agonist group (200μmol/L aluminum maltolate +100μmol/L mGluR1 agonist),and an aluminum maltolate +inhibitor group (200μmol/L aluminum maltolate +100μmol/L mGluR1 inhibitor),and all groups were treated for 24h. The mRNA expression levels of PKC and subunits of NMDAR in PC12 were measured by real-time fluorescence quantitative PCR, the protein expression levels of PKC and subunits of NMDAR by Western blot,and the PKC enzyme activity of PC12 by ELISA. [Results]Compared with the control group,the mRNA expression levels ofPKC,N.MDAR1,NMDAR2A,and NMDAR2B of the aluminum maltolate exposure group were decreased by 39%,21%,38%,and 26%,respectively (P<0.05),and the protein expression levels decreased by 39%,31%,41%,and 46%,respectively (P <0.05).The NMDAR1 mRNA and protein expression levels of the aluminum maltolate +inhibitor group were increased by 32% and 36%,respectively,compared with the aluminum maholate exposure group (P <0.05).The NMDAR2B mRNA expression levels of the aluminum maholate +inhibitor.group was decreased by 46%,and the NMDAR2B protein expression levels of the aluminum maltolate +agonist group was increased by 95%, compared with the aluminum maltolate exposure group (P<0.05).Compared with the control group,the PKC enzyme activity of the aluminum maltolate exposure group decreased by 56%,while the PKC activity of the aluminum maltolate+agonist group was upregulated by 116% compared with the maholate exposure group (P <0.05). [Conclusion ]The PKC and subunits of NMDAR mRNA and protein expressions i

关 键 词：麦芽酚铝 mGluR1激动 mGluR1抑制 PKC NMDAR 

分 类 号：R135[医药卫生—劳动卫生]