Experimental study on differential expression genes of glaucoma and potential intervention of traditional Chinese medicine based on bio-informatics
Objective To obtain the differential genes related to glaucoma pathogenesis by bio-informatics,and then through data mining,find out the traditional Chinese medicines that can interfere with the differenti-al genes in the treatment of glaucoma,and conduct in vitro experimental research to provide new research i-deas for traditional Chinese medicine treatment of glaucoma and experimental basis.Methods The dataset GSE27276 containing the whole-genome expression of glaucoma was obtained from the GEO database.Dif-ferential gene analysis was performed on it using R language.The protein-protein Interaction(PPI)analysis was performed by using String database.The core differential genes were screened by Cytoscape software.Using python language,the compounds corresponding to the core differentially expressed genes and related traditional Chinese medicines were captured in the TCMSP database,and then the most frequently occurring traditional Chinese medicines were selected for in vitro cell experimental research.Retinal ganglion RGC-5 cells were used in vitro cell experiments,and the cell model was created by hydrogen peroxide injury.MTT and MTS were used to screen the effective concentration of the drug,and the cell survival rate was detected after the injury model was administered.Western Blot was used to detect Bcl-2,Bax,the expression of caspase-3 protein,intracellular reactive oxygen species(ROS)were detected by chemical fluorescence meth-od,and the levels of tumor necrosis factor α(TNF-α)and interleukin(IL-1β)were detected by double-an-tibody sandwich method.Results A total of 194 differentially expressed genes were obtained from GSE27276,including 130 down-regulated genes and 64 up-regulated genes,and 15 up-regulated and 15 down-regulated core differentially expressed genes were obtained by Cytoscape.Using Python language,a to-tal of 6 048/frequency compounds were captured in the TCMSP database,and a total of 498 traditional Chi-nese medicines were finally obtained.The most frequent traditional Chinese medicine is wax gourd skin(2 852 frequency).The results of in vitro experiments show that using MTT and MTS methods to detect cell viability,high,medium and low concentrations of wax gourd peel water extract can improve the cell viability after hydrogen peroxide injury.The results of Western Blot detection showed that the water extract of wax gourd rind could up-regulate Bcl-2/Bax,and down-regulate the protein expressions of Bax and Caspase-3;The results of Ros test showed that compared with the hydrogen peroxide model group,the levels of ROS in the three concentration groups of wax gourd skin aqueous extract decreased significantly;The results of ELISA test showed that water extract of wax gourd peel could reduce the levels of TNF-α and IL-1β in in-jured cells.Conclusion Potential anti-glaucoma traditional Chinese medicines can be explored through bio-informatics methods.The experimental verification shows that the water extract of wax gourd peel has a pro-tective effect on retinal ganglion cells,and its mechanism may be related to anti-inflammatory and anti-oxi-dative damage.
glaucomabio-informaticstraditional Chinese medicineretinal ganglion cellscell experi-ments
万方数据
维普
