Abstract:

Objective To investigate the regulatory effect of Dendrobium nobile Lindl alkaloids（DNLA） on the proliferation， migration， and epithelial-mesenchymal transition （EMT） of HLEB3 cells induced by TGF-β2. Methods HLEB3 cells were cultivated in vitro and classified into the control group （DZ）， the model group （TGF-β2）， and the treatment group （TGF-β2 + DNLA）. TGF-β2 induced the EMT process of HLEB3 cells. Changes in cell morphology were observed through an inverted microscope. Cell proliferation， apoptosis， and migration were detected by CCK-8 assay， scratch test， and Transwell assay. The total RNAs of the samples were extracted for transcriptome analysis. Bioinformatics processing was employed to obtain relevant information on gene expression differences at the transcriptional level， biological processes， and related signaling pathways. The Western blot （WB） technique was utilized to detect EMT-related proteins to elucidate the mechanism of action of DNLA on lens epithelial cells. Results The study revealed that 10 μg/ml DNLA was suitable for the growth of HLEB3 cells. After 48 hours of the scratch test， the migration rate of the TGF-β2 + DNLA group significantly decreased （P < 0.01）. The Transwell results indicated that the cell migration ability of the TGF-β2 + DNLA group was notably weakened （P < 0.05）. Through bioinformatics， it was discovered that the prevention and treatment of posterior capsular opacification （PCO） by DNLA might be associated with cell junctions， cytoskeleton construction， and fibronectin binding. The pathogenesis of PCO may be related to multiple signaling pathways， including the TGF-β signaling pathway， TNF signaling pathway， and PI3K-Akt signaling pathway. Simultaneously， DNLA could reduce the expression levels of FNI， Smad2/3， and α-SMA proteins and increase the expression of E-cadherin protein. This indicates that DNLA can alleviate abnormal proliferation， migration， and EMT of lens epithelial cells by enhancing intercellular adhesion junctions and weakening cell migration ability， thereby playing a role in preventing and treating posterior capsular opacification. Conclusions DNLA can significantly inhibit the abnormal proliferation and migration of HLEB3 cells， alleviate the EMT process induced by TGF-β2， and prevent and control the occurrence of posterior capsular opacification and other ocular diseases. The mechanism of action might be related to the intervention of the TGF-β/smad signaling pathway and fibrosis proteins such as ZO-1 and E-cadherin.

Key words: Dendrobium nobile Lindl alkaloids, epithelial-mesenchymal transition, lens epithelial cell injury, α-SMA