[目的]研究氯化锰对人骨髓神经母细胞瘤细胞株(SH-SY5Y)线粒体损伤、氧化应激、多巴胺分泌及PARK2表达的影响。[方法]0、100、300、500μmol/L浓度氯化锰染毒SH-SY5Y细胞24 h后,用MTT法测细胞抑制率(反映线粒体损伤情况),石墨炉原子吸...[目的]研究氯化锰对人骨髓神经母细胞瘤细胞株(SH-SY5Y)线粒体损伤、氧化应激、多巴胺分泌及PARK2表达的影响。[方法]0、100、300、500μmol/L浓度氯化锰染毒SH-SY5Y细胞24 h后,用MTT法测细胞抑制率(反映线粒体损伤情况),石墨炉原子吸收光谱法测定细胞内锰浓度,高度水溶性四唑盐(WST-1)法测定细胞内超氧化物歧化酶(SOD)活性,硫代巴比妥酸法测定细胞内丙二醛(MDA)含量,反相高效液相色谱-荧光法测定细胞内多巴胺(DA)含量,实时荧光定量-PCR检测PARK2 m RNA表达,蛋白免疫印迹法检测Parkin蛋白表达。[结果]与对照组比较,MnCl2浓度为300、500μmol/L时,细胞抑制率(线粒体损伤)增高(P<0.01)。与对照组比较,染锰组细胞内锰浓度升高(P<0.05或P<0.01)。与对照组比较,MnCl2浓度为300、500μmol/L时,SOD活性和DA含量降低(P<0.01),MDA含量升高(P<0.01);细胞PARK2 m RNA表达和Parkin蛋白表达降低(P<0.01)。相关性分析显示,PARK2 m RNA表达与细胞抑制率(线粒体损伤)、细胞内锰浓度及MDA含量呈负相关,r值分别为-0.872、-0.880、-0.862(均P<0.01);PARK2 m RNA表达与SOD活性、DA含量以及Parkin蛋白表达呈正相关,r值分别为0.879、0.859、0.809(均P<0.01)。[结论]氯化锰暴露可引起SH-SY5Y细胞的线粒体损伤、氧化应激、DA分泌减少和PARK2表达下降。展开更多
The PARK2 gene is a common disease gene in Han Chinese patients with Parkinson＇s disease.The detection of mutations in the PARK2 gene remains low.To investigate the role PARK2 gene mutations play in the pathogenesis ...The PARK2 gene is a common disease gene in Han Chinese patients with Parkinson＇s disease.The detection of mutations in the PARK2 gene remains low.To investigate the role PARK2 gene mutations play in the pathogenesis of Parkinson＇s disease,30 Han Chinese patients with early-onset Parkinson＇s disease and 38 normal controls were studied to determine the sequence changes of 1,4,6 and 7 exon sections.In the 30 patients with Parkinson＇s disease,a heterozygous intron mutation（nt 119,G→G/A）in exon 1 was detected in one case;a homozygous intron mutation（nt 526500,T→C）between intron 3 and exon 4 in fourteen cases was found;a heterozygous intron mutation（nt 526607,G→G/A）between intron 3 and exon 4 was observed in eight cases;an exon 6missense mutation（nt 754317,C→C/T;codon 193,CGG→CGG/TGG;aa 193,Arg→Arg/Trp）in three cases was seen;and an exon 7 missense mutation（nt 941943,C→A/C;codon 272,CTC→CTC/ATC;aa 272,Leu→Leu/lle）was found in one case.These changes were not found in the normal population.The results indicated that the PARK2 exons 6 and 7 mutations are possibly pathogenic mutations,along with the intron 3-exert 4 and exon 1 mutations.PARK2 gene mutations are possible factors leading to the onset of Parkinson＇s disease.展开更多
文摘[目的]研究氯化锰对人骨髓神经母细胞瘤细胞株(SH-SY5Y)线粒体损伤、氧化应激、多巴胺分泌及PARK2表达的影响。[方法]0、100、300、500μmol/L浓度氯化锰染毒SH-SY5Y细胞24 h后,用MTT法测细胞抑制率(反映线粒体损伤情况),石墨炉原子吸收光谱法测定细胞内锰浓度,高度水溶性四唑盐(WST-1)法测定细胞内超氧化物歧化酶(SOD)活性,硫代巴比妥酸法测定细胞内丙二醛(MDA)含量,反相高效液相色谱-荧光法测定细胞内多巴胺(DA)含量,实时荧光定量-PCR检测PARK2 m RNA表达,蛋白免疫印迹法检测Parkin蛋白表达。[结果]与对照组比较,MnCl2浓度为300、500μmol/L时,细胞抑制率(线粒体损伤)增高(P<0.01)。与对照组比较,染锰组细胞内锰浓度升高(P<0.05或P<0.01)。与对照组比较,MnCl2浓度为300、500μmol/L时,SOD活性和DA含量降低(P<0.01),MDA含量升高(P<0.01);细胞PARK2 m RNA表达和Parkin蛋白表达降低(P<0.01)。相关性分析显示,PARK2 m RNA表达与细胞抑制率(线粒体损伤)、细胞内锰浓度及MDA含量呈负相关,r值分别为-0.872、-0.880、-0.862(均P<0.01);PARK2 m RNA表达与SOD活性、DA含量以及Parkin蛋白表达呈正相关,r值分别为0.879、0.859、0.809(均P<0.01)。[结论]氯化锰暴露可引起SH-SY5Y细胞的线粒体损伤、氧化应激、DA分泌减少和PARK2表达下降。
文摘The PARK2 gene is a common disease gene in Han Chinese patients with Parkinson＇s disease.The detection of mutations in the PARK2 gene remains low.To investigate the role PARK2 gene mutations play in the pathogenesis of Parkinson＇s disease,30 Han Chinese patients with early-onset Parkinson＇s disease and 38 normal controls were studied to determine the sequence changes of 1,4,6 and 7 exon sections.In the 30 patients with Parkinson＇s disease,a heterozygous intron mutation（nt 119,G→G/A）in exon 1 was detected in one case;a homozygous intron mutation（nt 526500,T→C）between intron 3 and exon 4 in fourteen cases was found;a heterozygous intron mutation（nt 526607,G→G/A）between intron 3 and exon 4 was observed in eight cases;an exon 6missense mutation（nt 754317,C→C/T;codon 193,CGG→CGG/TGG;aa 193,Arg→Arg/Trp）in three cases was seen;and an exon 7 missense mutation（nt 941943,C→A/C;codon 272,CTC→CTC/ATC;aa 272,Leu→Leu/lle）was found in one case.These changes were not found in the normal population.The results indicated that the PARK2 exons 6 and 7 mutations are possibly pathogenic mutations,along with the intron 3-exert 4 and exon 1 mutations.PARK2 gene mutations are possible factors leading to the onset of Parkinson＇s disease.
