LRRK2干扰对MPP+诱导PD细胞模型线粒体功能的影响

目的 探讨富亮氨酸重复激酶2(LRRK2)干扰对1-甲基-4-苯基吡啶离子(MPP+)诱导的帕金森病(PD)细胞模型线粒体功能及CaMKK-β/AMPK通路的影响.方法 采用MPP+诱导传代培养的SH-SY5Y细胞构建PD细胞模型.将造模的SH-SY5Y细胞随机分PD模型组、空载转染组(空载转染PD模型细胞)和siRN...

The effect of LRRK2 interference on mitochondrial function of PD cell model induced by MPP+

Objective To explore the regulatory effect of leucine-rich repeat kinase 2(LRRK2)on mitochondrial function in MPP+induced SH-SY5Y cell.Methods The Parkinson's disease(PD)cell model was constructed by MPP+induced SH-SY5Y cells,and randomly divided into a PD model group,a null carrier transfection group(PD model cell transfected with null carrier),and a siRNA transfection group(PD model cell transfected with siRNA-LRRK2).Untreated SH-SY5Y cell was set as the normal group.Each group had three replicates.The mRNA expression of LRRK2 was detected by qRT-PCR.The protein expression of LRRK2 was detected by Western blot.JC-1 probe was employed to measure mitochondrial membrane potential.The detection kits were used to detect ATP content and mitochondrial complex I(complex I)activity.Western blot was used to evaluate the expression of CaMKK-β/AMPK pathway related proteins.Results Compared with the normal group,the PD model group showed higher protein expression of LRRK2(P<0.05),lower mitochondrial membrane potential,reduced ATP content and complex I activity,and higher expression levels of CaMKK-β,AMPK and p-AMPKα(Thr 172)in CaMKK-β/AMPK pathway(P<0.05).Compared with the PD model group,while LRRK2 was disturbed by the transfection of siRNA-LRRK2-1,the mitochondrial membrane potential was elevated,the impaired mitochondrial function was partially recovered,the expressions of CaMKK-β/AMPK pathway related proteins were lowered significantly(P<0.05),and the phosphorylation level of AMPK was reduced significantly(P<0.05).Conclusions LRRK2 interference can regulate the mitochondrial functions of SH-SY5Y cells induced by MPP+,and inhibit the CaMKK-β/AMPK signal pathway.