MiR-29a Alleviates High Glucose-induced Inflammation and Mitochondrial Dysfunction via Modulation of IL-6/STAT3 in Diabetic Cataracts

Abstract

ABSTRACT Background: This in vitro 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. Methods: 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. Results: 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. Conclusions: 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.