SIRT3对大鼠糖尿病神经性疼痛的影响及机制研究-东南大学学报医学版

SIRT3对大鼠糖尿病神经性疼痛的影响及机制研究

单位：1. 南京医科大学康达学院附属涟水县人民医院, 江苏淮安 223400;
2. 江苏护理职业职业学院, 江苏淮安 223400;
3. 南京医科大学药学院, 江苏南京 211166

分类号：R966

出版年·卷·期（页码）：2025·44·第二期（263-271）

摘要：

目的:探讨沉默调节蛋白3(SIRT3)在糖尿病神经性疼痛(DNP)发生发展中的作用及其相关机制。方法:取80只大鼠,随机分为对照组、DNP组、SIRT3过表达组和SIRT3敲低组。SIRT3过表达组鞘内注射SIRT3慢病毒(LV-SIRT3),10 μl·d^-1;SIRT3敲低组鞘内注射SIRT3-siRNA慢病毒(LV-SIRT3-siRNA),10 μl·d^-1,两组均连续给药5 d。利用Von Frey和Hargreaves法进行大鼠行为学测定;免疫荧光双标染色法检测SIRT3细胞定位;Western blot、RT-qPCR检测SIRT3、锰超氧化物歧化酶(MnSOD)和过氧化氢酶(CAT)表达水平;相应试剂盒检测大鼠氧化应激水平,包括超氧化物歧化酶(SOD)、CAT、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-PX)和丙二醛(MDA)。结果:SIRT3主要定位于脊髓背角神经元。与对照组相比,DNP组SIRT3蛋白、mRNA表达及活性,大鼠机械缩足阈值(MWT)和热缩足潜伏期(TWL)明显降低(均P<0.01)。与DNP组相比,SIRT3过表达组SIRT3蛋白、mRNA表达及活性,MWT和TWL显著升高(均P<0.01)。与对照组相比,DNP组MnSOD、CAT蛋白和mRNA表达水平明显降低(P<0.01或P<0.05),氧化应激水平明显升高(P<0.01)。与DNP组相比,SIRT3过表达组MnSOD、CAT蛋白和mRNA表达水平显著升高(P<0.05或P<0.01),氧化应激水平明显降低(P<0.01)。与对照组相比,SIRT3敲低组大鼠SIRT3蛋白、mRNA表达及活性,MWT和TWL明显降低(P<0.01或P<0.05),MnSOD、CAT蛋白和mRNA表达水平均明显降低(均P<0.01),氧化应激水平明显升高(P<0.01)。结论:SIRT3可能通过调控抗氧化基因,改变机体氧化应激状态,参与DNP的发生发展,激活SIRT3可能作为一种潜在的DNP的治疗策略。

Objective: To investigate the role of silent regulatory protein 3(SIRT3) in the development of diabetic neuropathic pain(DNP) and its related mechanisms. Methods: Eighty rats were taken and randomly divided into control, DNP, SIRT3 overexpression and SIRT3 knockdown groups. The SIRT3 overexpression group was injected intrathecally with SIRT3 lentivirus vector(LV-SIRT3) 10 μl·d^-1, and the SIRT3 knockdown group was injected intrathecally with SIRT3-siRNA lentivirus(LV-SIRT3-siRNA) 10 μl·d^-1, and both groups were administered continuously for 5 d. Behavioral assays in rats were performed using the Von Frey and Hargreaves method; immunofluorescence double-label staining was used to detect the localization of SIRT3 cells; Western blot and RT-qPCR were used to detect the expression levels of SIRT3, manganese superoxide dismutase(MnSOD) and catalase(CAT); and the corresponding kits were used to detect the levels of oxidative stress in rats,such as SOD, CAT, glutathione(GSH), glutathione peroxidase(GSH-PX) and malondialdehyde(MDA). Results: SIRT3 was mainly localized in spinal dorsal horn neurons. Compared with the control group, SIRT3 protein, mRNA expression levels and activity, mechanical withdrawal threshold(MWT) and thermal withdrawal latency(TWL) were significantly decreased in the DNP group(all P<0.01). SIRT3 protein, mRNA expression levels and activity, MWT and TWL were significantly higher in the SIRT3 overexpression group compared with the DNP group(all P<0.01). MnSOD and CAT protein and mRNA expression levels were significantly lower(P<0.01 or P<0.05) and oxidative stress levels were significantly higher(P<0.01) in the DNP group compared with the control group. Compared with the DNP group, MnSOD and CAT protein and mRNA expression levels were significantly higher in the SIRT3 overexpression group(P<0.05 or P<0.01), and the level of oxidative stress was significantly lower(P<0.01). Compared with the control group, SIRT3 protein, mRNA expression levels and activity, MWT and TWL were significantly decreased in the SIRT3 knockdown group of rats(P<0.05 or P<0.01), and the levels of MnSOD and CAT protein and mRNA expression were significantly decreased(all P<0.01), the levels of oxidative stress were significantly elevated, and the difference was statistically significant(P<0.01). Conclusion: SIRT3 may be involved in the development of DNP by modulating antioxidant genes and altering the oxidative stress state of the body, and activation of SIRT3 may serve as a potential therapeutic strategy for DNP.

参考文献：

[1] XU D,MA X,SUN C,et al.Emerging roles of circular RNAs in neuropathic pain[J].Cell Prolif,2021,54(12):E13139.
[2] 蔡妙国,邵卫,俞慧君,等.SNI小鼠中调控ROS对疼痛和脊髓Pink1蛋白的影响及机制研究[J].中国临床药理学与治疗学,2019,24(11):1263-1268.
[3] TRECARICHI A,FLATTERS S.Mitochondrial dysfunction in the pathogenesis of chemotherapy-induced peripheral neuropathy[J].Int Rev Neurobiol,2019,145:83-126.
[4] ZHAO H,LUO Y,CHEN L,et al.Sirt3 inhibits cerebral ischemia-reperfusion injury through normalizing Wnt/β-catenin pathway and blocking mitochondrial fission[J].Cell Stress Chaperones,2018,23(5):1079-1092.
[5] 杨静,李民.SIRT3介导的线粒体功能和代谢调控机制在2型糖尿病中作用的研究进展[J].基础医学与临床,2021,41(12):1828-1832.
[6] 李鑫,李春喜.Sirtuins在2型糖尿病发病中的作用[J].生理科学进展,2020,51(5):377-384.
[7] 李永男,车守梅,张祁,等.米诺环素对神经病理性疼痛大鼠海马促炎性细胞因子表达的影响[J].医学研究杂志,2019,48(7):54-58.
[8] TIAN Y G,ZHANG J.Protective effect of SIRT3 on acute lung injury by increasing manganese superoxide dismutase-mediated antioxidation[J].Mol Med Rep,2018,17(4):57-65.
[9] DAI S H,CHEN T,WANG Y H,et al.SIRT3 attenuates hydrogen peroxide-induced oxidative stress through the preservation of mitochondrial function in HT22 cells[J].Int J Mol Med,2014,34:1159-1168.
[10] KITADA M,KUME S,KANASAKI K,et al.Sirtuins as possible drug targets in type 2 diabetes[J].Curr Drug Trgets,2013,14(6):622-636.
[11] ZHOU C H,ZHANG M X,ZHOU S S,et al.SIRT1 attenuates neuropathic pain by epigenetic regulation of mGluR1/5 expressions in type 2 diabetic rats[J].Pain,2017,158(1):130-139.
[12] 邓梦秋.Sirt3通过调控酮体代谢抑制小胶质细胞炎症反应参与神经病理性疼痛的机制研究[D].上海:中国人民解放军海军军医大学,2024.
[13] SHEN Y,WU Q,SHI J,et al.Regulation of SIRT3 on mitochondrial functions and oxidative stress in Parkinson's disease[J].Biomed Pharmacother,2020,132:110928.
[14] RODRIGUEZ R H,GARCIA G A,PICKETT C,et al.Compartmentalized oxidative stress in dopaminergic cell death induced by pesticides and complex I inhibitors:distinct roles of superoxide anion and superoxide dismutases[J].Free Radic Biol Med,2013,61:370-383.
[15] ZHANG H,ZHAO Z,PANG X,et al.MiR-34a/sirtuin-1/foxo3a is involved in genistein protecting against ox-LDL-induced oxidative damage in HUVECs[J].Toxicol Lett,2017,277:115-122.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录