Li Hua, Xu Lingxiao, Song Hui
Tianjin Eye Hospital, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin Eye Institute, Clinical College of Ophthalmology Tianjin Medical University, Tianjin, China.
Tianjin Eye Hospital, Nankai University Affiliated Eye Hospital, Tianjin, China.
Curr Eye Res. 2021 Sep;46(9):1325-1332. doi: 10.1080/02713683.2021.1887272. Epub 2021 Feb 21.
: This study was designed to reveal the role of miR-29a in high glucose-induced cellular injury through the modulation of IL-6/STAT3 in diabetic cataracts.: The expression of miR-29a and STAT3 in the lens capsules of patients with or without diabetes was determined by RT-PCR. The levels of the IL-6 proinflammatory cytokine in the aqueous humor were detected by ELISA. HLE B-3 cells were cultured in normal glucose (NG; 5 mM) or high glucose (HG; 40 mM). After transfection with miR-29a, si-STAT3, or a negative control vector, the levels of IL-6 and STAT3 were detected. A CCK-8 assay was used to determine cell viability. We used flow cytometry to assess changes in reactive oxygen species (ROS) production, mitochondrial membrane potential (MMP), and apoptosis induced by oxidative stress. Western blotting was used to determine the expression of the oxidative injury markers superoxide dismutase (SOD) and malondialdehyde (MDA) and the apoptosis markers Bcl-2 and Bax.: Reduced miR-29a, increased STAT3 expression, and IL-6 release were demonstrated in the lens capsules and aqueous humor of patients with diabetes. The stimulation of apoptosis and the loss of MMP induced by HG were attenuated by transfection with a miR-29a mimic and si-STAT3. ROS production, increased MDA content, decreased SOD activity, and upregulation of the apoptotic proteins Bcl-2/Bax were also partially alleviated by miR-29a overexpression, which shows their roles in oxidative injury. Furthermore, transfection with a STAT3 overexpression vector reversed the effects of miR-29a.: In conclusion, miR-29a mitigated HG-induced oxidative injury and exerted protective effects via IL-6/STAT3 signaling. Thus, miR-29a may be a potential therapeutic agent for diabetic cataracts.
本研究旨在通过调节糖尿病性白内障中白细胞介素-6（IL-6）/信号转导与转录激活因子3（STAT3）来揭示微小RNA-29a（miR-29a）在高糖诱导的细胞损伤中的作用。通过逆转录聚合酶链反应（RT-PCR）测定糖尿病患者和非糖尿病患者晶状体囊膜中miR-29a和STAT3的表达。采用酶联免疫吸附测定法（ELISA）检测房水中促炎细胞因子IL-6的水平。人晶状体上皮B-3（HLE B-3）细胞在正常葡萄糖（NG；5 mM）或高糖（HG；40 mM）条件下培养。用miR-29a、小干扰RNA-STAT3（si-STAT3）或阴性对照载体转染后，检测IL-6和STAT3的水平。采用细胞计数试剂盒-8（CCK-8）法测定细胞活力。我们使用流式细胞术评估活性氧（ROS）产生、线粒体膜电位（MMP）以及氧化应激诱导的细胞凋亡的变化。采用蛋白质免疫印迹法检测氧化损伤标志物超氧化物歧化酶（SOD）和丙二醛（MDA）以及凋亡标志物Bcl-2和Bax的表达。糖尿病患者的晶状体囊膜和房水中miR-29a表达降低、STAT3表达增加以及IL-6释放增加。用miR-29a模拟物和si-STAT3转染可减轻HG诱导的细胞凋亡和MMP丧失。miR-29a过表达也部分减轻了ROS产生、MDA含量增加、SOD活性降低以及凋亡蛋白Bcl-2/Bax上调，这表明它们在氧化损伤中的作用。此外，用STAT3过表达载体转染可逆转miR-29a的作用。总之，miR-29a减轻了HG诱导的氧化损伤，并通过IL-6/STAT3信号发挥保护作用。因此，miR-29a可能是糖尿病性白内障的一种潜在治疗药物。
