慢性病理性神经痛导致心肌线粒体功能障碍  

ALDH2 confers protection against chronic neuropathic pain-induced myocardial mitochondrial dysfunction

作　　者：杨京润 李晨[2] 薛涵 王一石 杨铮[2] 殷玥[1,2] 石曌玲[1,2,3] 余璐[4] 马恒[1,2] 

机构地区：[1]第四军医大学基础医学院生理学教研室,陕西西安710032 [2]第四军医大学基础医学院病理生理学教研室,陕西西安710032 [3]第四军医大学西京医院儿科,陕西西安710032 [4]第四军医大学基础医学院病理学教研室,陕西西安710032

出　　处：《心脏杂志》2017年第5期501-505,543,共6页Chinese Heart Journal

基　　金：国家自然科学基金项目资助(31671424;31571413;81322004)

摘　　要：目的探讨慢性神经性病理痛对心肌线粒体功能的影响。方法用10只C57雄性小鼠通过背根神经节(QRG)压迫3周方法建立慢性疼痛模型(Chronic compression of DRG,CCD)组;用10只小鼠做无痛对照(Con)组;另20只1月龄小鼠经尾静脉分别注射以9型腺相关病毒为载体的重组病毒,其中10只注射AAV9-CMV-ALDH2[3×1011viral genomes(vg)/只,病毒基因组]上调乙醛脱氢酶(ALDH)2的蛋白表达,剩余10只注射AAV9-CMVGFP(绿色荧光蛋白)为病毒对照。4周后,ALDH2稳定过表达之后,再建立CCD模型(CCD+ALDH2)组。慢性疼痛3周后提取心肌组织,用荧光探针检测心肌内活性氧自由基(ROS)的水平;用美国Sigma公司试剂盒测定丙二醛(MDA)的含量;用免疫共沉淀方法(Co-IP)测定心肌中锰超氧化物歧化酶(Mn SOD)的乙酰化水平和去乙酰化酶(SIRT)3的羰基化水平;用美国Cayman公司试剂盒检测Mn SOD的活性。结果慢性疼痛导致小鼠心肌氧化应激水平和脂质过氧化损伤显著增加,线粒体膜电位较对照组降低(P<0.05);慢性痛小鼠心肌中线粒体抗氧化酶Mn SOD乙酰化程度较对照组增加,活性减退(均P<0.05)。上述结果提示:慢性痛可导致心肌线粒体功能受损。进一步分析机制发现,慢性痛小鼠血浆中的活性羰基类物质4-羟基壬烯醛(4-HNE)较对照组显著增加(P<0.05),导致心肌线粒体去乙酰化酶SIRT3被羰基化修饰。上调ALDH2表达可有效的抑制慢性痛导致的心肌蛋白质羰基应激,减轻疼痛,有效保护心肌线粒体功能。结论慢性疼痛可诱发心肌羰基应激导致线粒体功能障碍,上调ALDH2可有效抑制疼痛并保护心肌线粒体功能。AIM To investigate the effects of chronic neuropathic pain （CNP） on the function of myocardial mitochondria. METHODS Forty male C57 mice were used in the present study. Ten mice were used to establish chronic neuropathic pain model using chronic compression of dorsal root ganglion （CCD）. Ten mice were regardedas control group （Con）. Ten mice of one month were injected with AAV9-CMV-ALDH2 （3×10^11 vg per mice） via vena caudalis. Ten mice of one month injected with AAV9-CMV-GFP via vena caudalis were regarded as the negative control. After 4 weeks, ALDH2 was over-expressed stably, and CCD model was established （CCD＋ALDH2）. After 3 weeks of CCD, the myocardial tissues were extracted. Detected the levels of reactive oxygen species （ROS）, malondialdehyde （MDA）, the activity and acetylation of MnSOD, and the activity and carbonylation of SIRT3. RESULTS CNP dramatically increased mitochondrial oxidative stress and lipid peroxidation, and decreased mitochondrial membrane potential in mice myocardium. The acetylation of MnSOD increased significantly in CNP mice （P〈0.05）, leading to the decreased activity （P〈0.05）. The results mentioned above suggested that CNP triggered mitochondrial dysfunction in the myocardium. Furthermore, the serum level of 4-hydroxynonenal （4-HNE） increased markedly （P〈0.05）, resulting in the carbonylation of SIRT3. The up-regulation of ALDH2 significantly attenuated CNP-induced carbonyl stress and pain （P〈0.05）, and maintained mitochondrial function. CONCLUSION CNP induces myocardial carbonyl stress and mitochondrial dysfunction. The up-regulation of ALDH2 effectively mitigates pain and myocardial mitochondrial dysfunction.

关 键 词：慢性病理性神经痛 乙醛脱氢酶2 去乙酰化酶3 线粒体 

分 类 号：Q463[生物学—生理学]