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MRL/lpr 狼疮小鼠线粒体细胞色素 C 氧化酶亚基 I 的表达特征。

The expression characteristics of cytochrome C oxidase subunit I in mitochondrial of MRL/lpr lupus mice.

机构信息

Department of Dermatology, The First Affiliated Hospital of Guangzhou Medical University, China.

Department of Dermatology, The First Affiliated Hospital of Sun Yat-sen University, China.

出版信息

Clin Exp Rheumatol. 2021 Jan-Feb;39(1):44-51. doi: 10.55563/clinexprheumatol/qd9bag. Epub 2020 Mar 28.

Abstract

OBJECTIVES

We sought to analyse the expression characteristics of cytochrome C oxidase subunit I in mitochondrial of MRL/lpr lupus mice.

METHODS

The whole blood of MRL/lpr lupus mice was detected for whole mitochondrial genome sequencing performed by Illumina HiSeq PE150 instrument, compared with house mouse (NC_005089.1) and screened for the maximum difference gene, MT-CO1. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot were used to detect the mRNA and protein expression of MT-CO1 in lupus mice and control mice. The total antioxidant capacities of lupus mice and control mice were measured using the rapid 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) method.

RESULTS

The mitochondrial genome sequencing showed that five mitochondrial genes had base differences and MT-CO1 was the maximum difference gene, 31 in total. Among the 31 base difference sites, 2 were missense mutations and 29 were synonymous_variant. qRT-PCR test results showed that the MT-CO1 expression in lupus mouse blood was statistically lower than that in control mice blood (t=4.333; p=0.0003). Western blot test results revealed that the expression of MT-CO1 was lower in the lupus mice compared with the control mice at the protein level. Serum total antioxidant capacity testing showed that: the serum total antioxidant capacity of lupus mice was statistically lower than that of the control mice (t=9.957; p<0.0001).

CONCLUSIONS

High mutation rate and decreased expression of MT-CO1 in MRL/lpr lupus mice accompanied the decrease of antioxidant capacity, which indicated that abnormal MT-CO1 might be involved in the pathogenesis of SLE and the production of anti-dsDNA antibodies.

摘要

目的

分析 MRL/lpr 狼疮小鼠线粒体细胞色素 C 氧化酶亚基 I 的表达特征。

方法

采用 Illumina HiSeq PE150 测序仪对 MRL/lpr 狼疮小鼠全血进行全线粒体基因组测序,与小家鼠(NC_005089.1)进行比较,筛选出最大差异基因 MT-CO1。采用实时荧光定量聚合酶链反应(qRT-PCR)和蛋白质印迹法检测狼疮小鼠和对照小鼠 MT-CO1 的 mRNA 和蛋白表达。采用快速 2,2'-偶氮双(3-乙基苯并噻唑啉-6-磺酸)(ABTS)法测定狼疮小鼠和对照小鼠的总抗氧化能力。

结果

线粒体基因组测序显示,有 5 个线粒体基因碱基存在差异,其中 MT-CO1 是最大差异基因,共有 31 个。在 31 个碱基差异位点中,2 个为错义突变,29 个为同义变异。qRT-PCR 检测结果显示,狼疮小鼠血液中 MT-CO1 的表达明显低于对照小鼠(t=4.333;p=0.0003)。Western blot 检测结果显示,狼疮小鼠 MT-CO1 的表达水平明显低于对照小鼠。血清总抗氧化能力检测显示:狼疮小鼠的血清总抗氧化能力明显低于对照小鼠(t=9.957;p<0.0001)。

结论

MRL/lpr 狼疮小鼠 MT-CO1 突变率高、表达降低,伴随抗氧化能力降低,提示异常 MT-CO1 可能参与 SLE 的发病机制和抗 dsDNA 抗体的产生。

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