A Comparative Study On The Inhibitory Effects Of Commonly Used Clinical Drugs On In Vitro Fibroblasts From Recurrent Pterygium

Objective1.Detection of DXM,IFN?-2b,MMC,5-FU,Cs A and FK506 the six clinical commonly used drugs for in vitro culture of recurrent pterygium fibroblast half inhibitory concentration.2.This study was to compare the efficacy under half inhibitory concentration of DXM,IFN?-2b,MMC,5-FU,Cs A and FK506 on proliferation and apoptosis of recurrent pterygium-derived fibroblasts in vitro.Methods Part1 The specimens of recurrence pterygium were collected during surgery in Eye Hospital of Tianjin Medical University from May 2015 to July 2017 under the writern informed consen.Fibroblasts were isolated and cultured by explant culture method and identified by immunochemistry.Part2 DXM,IFN-?2b,MMC,5-FU,Cs A and FK506 were added into the medium for48 hours,respectively,and the cells cultured without drug were used as the control group.The inhibitory efficency of different concentrations of ethanol extract of DXM,IFN-?2b,MMC,5-FU,Cs A and FK506 on the cell proliferation was assayed by cell cunting kit-8(CCK-8),and 50% inhibiting concentration(IC50)of the drugs was calculated.Part3 The cells were treated by the IC50 dose of drugs for 48 hours,and cell apoptotic proportion and cell cycle were assessed by flow cytometry analysis.The expression of proliferating cell nuclear antigen(PCNA)in the cells after treated by drugs was detected by immunochemistry.Results Cultured cells grew well with the fusiform shape and radial arrangement.Vimentine showed the positive expression and keratine was absently expressed in the cells.The IC50 to the cells was 3.5×103 mg/,6.1×102 mg/L,3.2×10-1 mg/L,2.2×101mg/L,6.3×101 mg/L and 6.0×101 mg/L in the DXM,IFN?-2b,MMC,5-FU,Cs A and FK506,respectively.In the 48 hours after treated by the IC50 drugs,the apoptotic ratio was(35.00±3.21)%,(30.37±1.67)%,(26.11±0.75)%,(22.01±0.07)%,(20.95±1.68)% and(19.85±0.52)% in the IFN?-2b group,Cs A group,MMC group,FK506 group,DXM group and 5-FU group,which was significantly higher than(11.38±2.18)% in the control group,showing a significant difference among the groups(F=63.73,P<0.01).The cell proporation of G0/G1 phase,S phase and G2/M phase was(85.64±2.62)%,(5.29±1.56)% and(2.73±2.66)% in the control group,and the cell proporation of G0/G1 phase was reduced,while that of S phase or G2/M phase was considerably increased in various drug groups(all at P<0.05),with the blocking efficency of cell cycle was in turn MMC,Cs A,5-FU,DXM,IFN-?2b and FK-506.The expressional rate of PCNA in the cells was(95.00±2.00)%,(82.67±5.04)%,(80.00± 2.78)%,(64.00±6.55)%,(38.00±3.00)%,(32.00±4.36)% and(29.67±3.02)% in the control group,FK506 group,DXM group,5-FU group,IFN-?2b group,Cs A group and MMC group,showing a significant difference among the groups(F=25.995,P<0.01),and the expressional rate of PCNA was significant lower in various drug group than that in the control group(all at P<0.05).Conclusions DXM,IFN-?2b,MMC,5-FU,Cs A and FK-506 are able to inhibit the the proliferation and promote the apoptosis of recurrent pterygium-derived fibroblasts in vitro,and MMC and Cs A appear to have a stronger effiency.