α-硫辛酸通过抑制小鼠miR-29b表达抑制氧化应激并促进糖尿病伤口愈合的作用

[EFFECT OF α-LIPOIC ACID IN INHIBITING OXIDATIVE STRESS AND PROMOTING DIABETIC WOUND HEALING BY SUPPRESSING EXPRESSION OF miR-29b IN MICE].

作者信息

Wu Jun, Tang Huiqin, Liu Qun, Gan Dingyun, Zhou Man

机构信息

Department of Endocrinology, the Third Hospital of Wuhan, Wuhan Hubei, 430060, P. R. China.

Department of Medical Administration, the Health and Family Planning Commission of Hubei Province.

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2016 Aug 8;30(8):1034-1037. doi: 10.7507/1002-1892.20160207.

DOI:10.7507/1002-1892.20160207

PMID:29786237

Abstract

OBJECTIVE

To investigate the effect of α-lipoic acid on the oxidative stress of wound tissues and diabetic wound healing in mice with diabetic feet.

METHODS

Sixty male C57BL/6J mice weighting 200-300 g were randomly divided into model group (control group, =15), α-lipoic acid-treated model group (=15), miR-29b mimic group (=15), and miR-29b mimic negative control group (NC group, =15). All animals received intraperitoneal injection of streptozocin to establish the diabetic model. Then, a full thickness wound of 5 mm×2 mm in size was created at 4 weeks after modeling. All mice were administrated with high-sugar-fat-diet. At the same day after modeling, α-lipoic acid-treated model group was continuously given intravenous injection of 100 mg/(kg·d) α-lipoic acid for 14 days; miR-29b mimic group and NC group received the tail intravenous injection of lentiviral vector for miR-29b mimic and miR-29b mimic negative control (a total of 2×10 TU), respectively, with the treatment of α-lipoic acid. The wound healing was observed and wound area was measured at 7 and 14 days. The wound tissues were harvested to detect the levels of superoxide dismutase (SOD) and glutathione (GSH) using xanthine oxidase method and 5, 5-dithiobis-2-nitrobenzoic acid staining method at 14 days. At the same day, 7, and 14 days after modeling, the relative miR-29b expression in wound tissues from control and α-lipoic acid-treated model groups was detected by real-time fluorescence quantitative PCR.

RESULTS

All mice survived to the experiment end. The wound healing was faster in α-lipoic acid-treated group than control group. At 7 and 14 days, the relative wound area and miR-29b expression level were significantly lower, while the contents of SOD and GSH were significantly higher in α-lipoic acid-treated group than control group (<0.05). In addition, miR-29b mimic group had significantly increased relative wound area and significantly decreased the contents of SOD and GSH when compared with NC group at 7 and 14 days (<0.05).

CONCLUSIONS

α-lipoic acid could inhibit oxidative stress and promote diabetic wound healing by suppressing expression of miR-29b in mice.

摘要

目的

探讨α-硫辛酸对糖尿病足小鼠伤口组织氧化应激及糖尿病伤口愈合的影响。

方法

将60只体重200 - 300 g的雄性C57BL/6J小鼠随机分为模型组（对照组，n = 15）、α-硫辛酸处理模型组（n = 15）、miR-29b模拟物组（n = 15）和miR-29b模拟物阴性对照组（NC组，n = 15）。所有动物均腹腔注射链脲佐菌素建立糖尿病模型。然后，在建模后4周制作一个大小为5 mm×2 mm的全层伤口。所有小鼠均给予高糖高脂饮食。建模后当天，α-硫辛酸处理模型组连续14天静脉注射100 mg/(kg·d)α-硫辛酸；miR-29b模拟物组和NC组分别尾静脉注射miR-29b模拟物和miR-29b模拟物阴性对照的慢病毒载体（共2×10 TU），并给予α-硫辛酸治疗。在第7天和第14天观察伤口愈合情况并测量伤口面积。在第14天收集伤口组织，采用黄嘌呤氧化酶法和5,5-二硫代双-2-硝基苯甲酸染色法检测超氧化物歧化酶（SOD）和谷胱甘肽（GSH）水平。在建模后当天、第7天和第14天，采用实时荧光定量PCR检测对照组和α-硫辛酸处理模型组伤口组织中miR-29b的相对表达。

结果

所有小鼠均存活至实验结束。α-硫辛酸处理组伤口愈合较对照组快。在第7天和第14天，α-硫辛酸处理组的相对伤口面积和miR-29b表达水平显著低于对照组，而SOD和GSH含量显著高于对照组（P < 0.05）。此外，在第7天和第14天，与NC组相比，miR-29b模拟物组的相对伤口面积显著增加，SOD和GSH含量显著降低（P < 0.05）。