alexa Protective effect of Fufang Xueshuantong on human retinal pigment epithelium cells injured by tert-butyl hydroperoxide in vitro
Ophthalmology

Ophthalmology

Journal of Clinical & Experimental Ophthalmology

Author(s): CHEN Xiaoyun, XIAO Wei, LUO Yan, MA Wei, LI Tao

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Objective To investigate the protective effect ofFufang Xueshuantong on human retinal pigment epithelium (hRPE) induced by tert-butyl hydroperoxide (t-BHP) in vitro. Methods hRPE cells were isolated from human eyes and cultured. The third to fifth passage were used in the following experiments. hRPE cells were exposed to 500μ M t-BHP and treated with various concentrations of Fufang Xueshuantong simultaneously. MTT assay was used to evaluate the cell viability to determine the optimal concentration of Fufang Xueshuantong. MTT assay and Annexin V/PI staining were used to compare the protective effect of Fufang Xueshuantong and each simple recipe in its composition on t-BHP-injured hRPE cells. hRPE cells were divided into three groups, including normal control group, t-BHP group and t-BHP with Fufang Xueshuantong group.Changes of cells and nuclei morphology were observed under a phase contrast microscope and a fluorescence microscope after Hoechst 33258 staining. The overproduction of ROS and change of mitochondrial membrane potential were assessed by MitoSOX staining and JC-1 staining. The hRPE-secreted VEGF was detected by ELISA kit. Results Exposured to 500μ M t-BHP for 6 hours resulted in the cells viability decreased to 49.9± 6.3%. While the cells viability of Fufang Xueshuantong group at different concentrations was higher than the t-BHP group. 0.25mg/ml Fufang Xueshuantong showed maximally protective effect on hRPE induced by t-BHP. The results of cells viability and apoptosis showed that the protective effect of Fufang Xueshuantong was much better than each simple recipe. Compared to t-BHP group, the application of Fufang Xueshuantong remarkably decreased dead cells and apoptotic nuclei, reduced the level of ROS production in mitochondria,maintained the mitochondrial membrane potential and inhibited the RPE-secreted VEGF induced by t-BHP.Conclusions This study suggests that Fufang Xueshuantong can protect hRPE cells against oxidative injury-induced cell death by reducing ROS in mitochondria, maintaining the mitochondrial membrane potential,inhibiting RPE from secreting VEGF.

This article was published in CHINESE JOURNAL OF PRACTICAL OPHTHALMOLOGY and referenced in Journal of Clinical & Experimental Ophthalmology

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