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A Study of the Mitochondrial Function of mtDNA Haplogroup D4 and B4a Cells and the Mechanism of These Haplogroups Effects on the Occurrence of Aging
Zhou Chao1,2, Gao Jing1, Xiong Jingting1, Qiu Ruyi1, Shen Lijun1, Lü Jianxin1,3*
1Attardi Institute of Mitochondrial Biomedicine, Zhejiang Provincial Key Laboratory of Medical Genetics, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou 325035, China; 2Children’s Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China; 3Hangzhou Medical College, Hangzhou 310053, China
Abstract: The aim of this study was to analysis the mitochondrial function in mitochondrial DNA (mtDNA) haplogroup D4 and B4a cell lines and to investigate the possible mechanism of aging. Haplogroup D4 was found to be significantly related to resistance to aging and haplogroup B4a was found to be a risk factor for aging. The mtDNA haplogroup D4 and B4a transmitochondrial cybrids with the same nuclear background were obtained bycytoplasmic hybrid. RT-PCR， clark electrode and gradient blue native polyacrylamide gelelectrophoresis (BN-PAGE)， TMRM and DCFH-DA detained， were carried out to examine mtDNA content and mtDNA transcription level， the oxygen consumption rate and mitochondrial complex expression， the mitochondrial membrane potential and reactive oxygen species (ROS) generation， in those cells. Results showed that the mitochondrial oxygen consumption rate， mitochondrial membrane potential and mtDNA (ND1， 7S RNA) transcription level of haplogroup D4 cybrids cell were significantly higher than those of haplogroup B4a cybrids cell， but ROS generation level was significantly lower than that of haplogroup B4a cybrids cell， although there are no significant on mtDNA content level between the two cells. In addition， the mitochondrial function was improved and oxidative damage was decreased in haplogroup D4 cybrids cell by promoting the expression of mtDNA and mitochondrial respiratory complex， so as to delay the occurrence of aging.
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