Effect Of Qufeng Huoxue Pill Containing Serum On Apoptosis And Expression Of Fas And FasL In Rat RPE Cells Induced By IRBP

Objective:To explore the functions of promoting blood pill containing medicine serum of IRBP intervened rat retinal pigment epithelial cell apoptosis inhibition,observe the functions of huoxue pill containing medicine serum of IRBP intervened the influence of the expression of Fas and Fas L RPE cells of rats.From the Angle of cell biology to investigate the functions of the mechanism of action of huoxue pill in the treatment of uveitis,for the wind to invigorate the pill in the treatment of uveitis provide experimental basis.Methods:(1)Choose healthy adults without eye disease Lewis,in the male rat enzyme digestion method for the generation of RPE cells,to the original RPE cells,the generation of develop and extend the observation of cell growth condition,the Cytokeratin-8 specificity antibodies immunofluorescence method for identification of cells.(2)Choose adult health rats only 40,male and female unlimited,weight about 240 g,randomly divided into blank group,the functions of promoting blood pill 2.5 times group(low dose group),the functions of promoting blood shot five times group(middle dose group)and the functions of 10 times huoxue pill group(high dose group),a total of four groups,respectively with distilled water and the functions of huoxue pill slurry to fill the stomach,2 times/day,7 days after lavage,abdominal aorta in blood,blank serum and high,medium and low functions of huoxue pill containing medicine serum.(3)Concentration of CCK 8 method to join different functions of huoxue pill,blank serum medicated serum cultured RPE cells of rats and not dealt with absorbance value of RPE cells in rats,observe the different concentration of medicated serum influence on RPE cell activity of rats,to choose the best effect on serum concentrations of RPE cells in rats.(4)The experimental group was divided into blank control group,model group 2,model group IRBP stimulated RPE cells,simulated in vitro uveitis environment,ELISA group detection of each group of RPE cells cytokine IL-6 before intervention and 6h after intervention,12 h,24h,36 h,48h.(5)The experiment is divided into blank control group,normal serum group,model group,treatment group 4,the current method to detect each group RPE cells apoptosis rate of rats,observe the functions of huoxue pill containing medicine serum of IRBP intervention under the influence of RPE cells apoptosis rate of rats.(6)The experiment is divided into blank control group,normal serum group,model group,treatment group 4,ELISA,PCR method to detect each group rats Fas,Fas L protein and m RNA expression in RPE cells,examination functions of huoxue pill after medicated serum Fas,Fas L expression of RPE cells in rats.Results:(1)Successfully cultivate RPE cells of rats.(2)The successful preparation of medicated serum,choose five times as experiment the intervention of the medicated serum concentration.(3)The rate of cell apoptosis was detected by flow cytometry,normal serum group and blank control group the apoptosis rate of RPE cells was essentially flat,there was no statistically significant difference(P > 0.05);model group,treatment group were compared with control group,the apoptosis rate of RPE cells increased significantly,the difference was statistically significant(P < 0.01);the treatment group compared with the model group,the apoptosis rate of RPE cells decreased significantly,the difference was statistically significant(P < 0.01).(4)ELISA method to detect content of Fas and Fas L protein groups of cells,normal serum group and blank control group differences had no statistical significance(P>0.05);Model group,treatment group compared with blank control group RPE cells of Fas and Fas L protein expression were significantly increased,there are significant difference statistically significant(P<0.01);The treatment group compared with model group RPE cells of Fas and Fas L protein expression significantly reduced,all have significant difference(P<0.01).(5)PCR method to detect the contents of Fas and Fas L m RNA,blank control group compared with normal serum group there was no statistically significant difference(P>0.05);Compared with blank control group,model group of Fas and Fas L m RNA difference have significant statistical significance(P<0.01),treatment group Fas m RNA difference was statistically significant(P < 0.05),treatment group Fas L m RNA difference have significant statistical significance(P<0.01);Compared with model group,treatment group of Fas and Fas L m RNA difference have significant statistical significance(P<0.01).Conclusions:Functions of huoxue pill containing medicine serum can inhibit IRBP under the action of RPE cells apoptosis in rats.Its mechanism is one of the functions of huoxue pill can inhibit the expression of Fas and Fas L protein,lower the Fas/Fas L constitute ratio.