Inhibition of NLRP3 Protects Human Lens Epithelial Cells against Oxidative Stress-Induced Apoptosis by NF-κB Signaling

Abstract

<b><i>Backgrounds/Aims:</i></b> To explore whether NLRP3 is involved in the development of cataract and to study the effect of NLRP3 on hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>)-induced injury in human lens epithelial cells. <b><i>Methods:</i></b> Oxidative stress-induced apoptosis model was constructed by treating HLEB3 cells with 50 µM H<sub>2</sub>O<sub>2</sub> at different times (6 h, 12 h) and was confirmed by flow cytometry and Western blot. HLEB3 were divided into NC, NC+H<sub>2</sub>O<sub>2</sub>, shNLRP3, and shNLRP3+H<sub>2</sub>O<sub>2</sub> groups. Quantitative real-time polymerase chain reaction and Western blot were employed to detect mRNA and protein expressions, DCFH-DA to measure reactive oxygen species production, and Annexin V-FITC/PI staining to determine cell apoptosis. <b><i>Results:</i></b> NLRP3 expression significantly increased in H<sub>2</sub>O<sub>2</sub>-induced HLEB3 cells. shRNA interference of NLRP3 inflammasome protects HLEB3 cells against oxidative stress-induced apoptosis by decreasing the expression levels of caspase-3 and Bax and increasing Bcl-2 expression. shNLRP3 was able to effectively suppress H<sub>2</sub>O<sub>2</sub>-induced apoptosis via inhibition of NF-κB signaling. <b><i>Conclusion:</i></b> NLRP3 might be involved in the apoptosis of lens epithelial cells. The inhibition of NLRP3 obviously attenuated H<sub>2</sub>O<sub>2</sub>-induced oxidative stress injury of human lens epithelial cells via NF-κB signaling.

RNA sequencing and bioinformatics analysis of human lens epithelial cells in age-related cataract

“…Yan et al found that the increased expression of laminin α4 in the anterior lens capsule is the main mechanism leading to the occurrence of ARC [16], which is consistent with our transcriptome sequencing results. Moreover, the inhibition of NLRP3 was shown to reduce oxidative stress-induced damage to human lens epithelial cells, while NLRP3 protein expression was significantly increased in lens epithelial cells under oxidative stress [17], which was also consistent with our verification results. However, more than 80% of the genes identified by transcriptome sequencing in this study have not previously been associated with ARC, so should be explored in future in-depth studies.…”

Echinatin mitigates H2O2-induced oxidative damage and apoptosis in lens epithelial cells via the Nrf2/HO-1 pathway

“…[3][4][5] Studies have confirmed that cataracts are directly related to the apoptosis of LECs caused by oxygen free radicals. 6,7 Thus, it is important to explore antioxidant drugs that can prevent the formation and development of cataracts.…”

Astragaloside IV alleviates oxidative stress‑related damage via inhibiting NLRP3 inflammasome in a MAPK signaling dependent pathway in human lens epithelial cells

“…AS‐IV possesses many pharmacological activities, such as antioxidative stress, anti‐inflammation via regulating various signaling pathways (L. Li et al, 2017). For instance, AS‐IV inhibited inflammatory response and oxidative stress via inhibition of MAPK signaling pathway (Hsieh et al, 2020), and AS‐IV also exhibited inhibitory effect on NLRP3 inflammasome (Leng et al, 2019; Xie et al, 2020), which protected HLECs from H 2 O 2 ‐induced apoptosis and inflammatory response via inhibition of NF‐κB signaling pathway (Zou et al, 2020).…”

“…AS‐IV protected against retinal iron overload toxicity through regulation of iron activity and inhibition of NF‐κB and MAPKs signaling activation (Song et al, 2021). Various studies have shown that AS‐IV exhibited suppressive effect on NLRP3 inflammasome (Leng et al, 2019; Xie et al, 2020), and inhibition of NLRP3 was able to protect HLECs from H 2 O 2 ‐induced apoptosis and inflammatory response by inhibiting the NF‐κB signaling pathway (Zou et al, 2020).…”