Effects Of Rho-kinase Inhibitor Fasudil On The Proliferation, Migration And Extracellular Matrix Synthesis Of Human Lens Epithelial Cells

BackgroundThe residual lens epithelial cells (lens epithelial cells, the LECs) of lens capsuleafter cataract surgery proliferate migration collagen deposition and lens fibersregenerated lead to posterior capsular opacity (posterior capsular opacification,PCO).PCO can cause eyesight drops again after cataract surgery in patients of one ofthe most common long-term complications. Mechanical trauma in cataract surgeryand postoperative inflammation, etc all can cause a lot of the secretion and release ofcytokines in aqueous humor, these cytokines as extracellular cues with receptors onthe surface of the lens epithelial cell membrane causes the cell signal transductionand gene transcription activation in the nuclei, thus inducing cell changes, eventuallylead to the formation of PCO.In this process, the intracellular signaling pathwaysplay an important role in the formation of PCO.Rho (Ras homologue) is actincytoskeleton reassemble, one of the main regulatory factor in cell signal transductionpathway plays a signal converter, or the role of molecular switch. Related studieshave shown that Rho can activate the key of the downstream Rho kinase effect oftarget molecules regulate a variety of biological behaviour of lens epithelial cells, butthe Rho/Rho kinase signaling pathway in the process of the formation of PCO role seldom reported. fasudil is currently used in clinical and commonly used in theexperiment of Rho/Rho kinase signaling pathway of selective inhibitors, it can notonly inhibit the activity of intracellular Ca2+, and can inhibition of intracellularmyosin light chain (MLC) phosphorylation levels, so that inhibiting function ofcytoskeleton. But the effect of fasudil on the mechanism of action of lens epithelialcells at present is less reported.ObjectiveThis experiment by using different concentration of fasudil to intervention tocultivate human lens epithelial cells in vitro, different point in time, respectively toobserve its effect on cell proliferation, migration and extracellular matrix synthesisaspects, and to explore its mechanism.MethodsCultivate human lens epithelial cells in vitro, divided them into the followingfive groups: control group drug treatment group, drug treatment group respectivelyfor,10μmol/L、20μmol/L、40μmol/L、60μmol/L, CCK-8method is used to detect cellproliferation at12,24and48h, Transwell Chambers migration experiment test cellmigration ability, ELISA method to detect each cell supernatant of type I collagen(COL I) fiber link protein (FN) and α-SMA expression in the cell.ResultsAccording to CCK-8, after the intervention of cell proliferation is restrained, theobvious concentration-dependence and time-dependence (P<0.05) TranswellChambers migration experiments have shown that48hours after the control cellsthrough the polycarbonate membrane is (29.20±1.28), the drug treatment group (theconcentrate of fasudil:10μmol/L、20μmol/L、40μmol/L、60μmol/L) number of cellmigration, respectively (24.40±1.33、17.00±1.10、14.60±0.68、6.60±1.29), withdifferent concentrations of fasudil, HLE-B3cells migration has inhibitory effect (P ＜0.05) in different concentrations of fasudil. In different concentrations of fasudilintervention group of that the fiber link protein (FN) in cells supernatant andcollagen type I (COL I) content and the expression of α-SMA significantly lowerthan the control group（P＜0.05）.Conclusion1. Cultivate human lens epithelial cells in vitro HLE-B3express a certainamount of COL-I、FN and α-SMA.2. Rho/Rho kinase signal transduction pathway inhibitor fasudil, can inhibitmigration in vitro cultivation of human lens epithelial cells, and can also inhibit theproliferation and the secretion of extracellular matrix synthesis...