| Objective:To explore the protective effect of Baicalin on hypoxia-reoxygenation injury of JEG-3 trophoblast cells and its mechanism,and to provide a theoretical basis for the intervention treatment of baicalin in preeclampsia.Method:The JEG-3 cell line purchased from the Chinese Academy of Sciences Cell Bank was cultured in vitro and the cells were divided into control group,hypoxia/reoxygenation group(H/R)and H/R+low concentration baicalin intervention group(LB).Group),H/R+ medium baicalin intervention group(MB group),H/R+high concentration baicalin group(HB group)(1.5625ug/ml,3.125ug/ml,6.25ug/ml),hypoxia 8h,reoxygenation 16 h PE cell model was prepared,cell viability was measured by CCK-8 assay,concentration of appropriate drug was calculated,cell apoptosis was detected by flow cytometry,Western blotting analysis Expression levels of BCL-2,XIAP,Caspase-3 and Caspase-9 in JEG-3 trophoblast cells.The data were analyzed by SPSS22.0 software.The results were expressed as mean±standard deviation.Two independent normal samples were compared using t-test,and the mean of samples was compared using F-test and LSD-t test,? = 0.05 for the test level,P <0.05 for the difference was statistically significant.Result:1.The flow cytometry results showed that the apoptosis rate of the control group,H/R,H/R+LB,H/R+MB,H/R+HB groups was [(2.09 ± 0.15)VS(20.77 ±0.95)VS(18.06 ± 0.23)VS(14.69 ± 0.37)VS(9.36 ± 0.31),F=687.85,P<0.01].Comparison of apoptosis rate in each group: compared with the the control group,H/R group apoptosis rate was increased,the difference was statistically significant,the t value was-30.61,and the P value was less than 0.01.Compared with group H/R,the apoptosis rates of H/R+LB,H/R+MB,and H/R+HB groups were all decreased,and the differences were statistically significant,the values of t were 5.02,8.53,and22.74 respectively,and the P values were all less than 0.05.Apoptosis rate of H/R+LB,H/R+MB,and H/R+HB group,F=605.04,P<0.01,the difference was statistically significant: between H/R+LB and H/ R+MB t=19.45,P<0.01,between H/R+MB and H/R+HB,t=23.88,P<0.01,between H/R+LB and H/R+HB,t=147.47,P< 0.01.2.Western bloting display(1)Bcl-2 expression level in control group,H/R,H/R+LB,H/R+MB,H/R+HB group[(0.71±0.03)VS(0.27±0.02)VS(0.36±0.03)VS(0.49±0.02)VS(0.53±0.03),F=161.12,P<0.01].Comparison of expression levels of Bcl-2 in eachg group:compared with the control group,the expression of Bcl-2 in the H/R group decreased,and the difference was statistically significant,the t value was 49.51,and the P value was less than 0.01.Compared with the H/R group,the expression of Bcl-2 in the H/R+LB,H/R+MB,and H/R+HB groups was all increased,the difference was statistically significant,the t values were-9.83,-65.00,and-39.50,respectively,and the P values were all less than 0.05.Bcl-2 expression levels in H/R+LB,H/R+MB,H/R+HB groups were compared,,F=44.37,P < 0.01,there was significant difference:between H/R+MB and H/R+LB,t=-22.52,P < 0.01,between H/R+MB and H/R+HB,t=-14.00,P=0.053,there was no significant difference,between H/R+HB and H/R+LB,t=-53.00,P < 0.01.(2)The expression of Caspase-3 in control group,H/R,H/R+LB,H/R+MB,H/R+HB group [(0.13 ± 0.01)VS(0.49 ± 0.02)VS(0.36 ± 0.01)VS(0.31 ±0.01)VS(0.21 ± 0.01),F=384.20,P<0.01.Comparison of expression levels of Caspase-3 in eachg group: compared with the control group,the expression of Caspase-3 in the H/R group increased,and the differences were statistically significant,the t value was-54.50,and the P value was less than 0.01.Compared with H/R group,the expression of Caspase-3 in H/R+LB,H/R+MB,and H/R+HB groupswere all decreased,and the differences were statistically significant,the t values were22.52,31.18,and 32.13,respectively.P values were all less than 0.01.Caspase-3expression levels in H/R+LB,H/R+MB,H/R+HB groups were compared,F=235.86,P<0.01,the difference was statistically significant: between H/R+ LB and H/R+MB,t=?,P<0.01,between H/R+MB and H/R+HB,t=31.00,P<0.01,between H/R+LB and H/R+HB,t= 46.00,P<0.01.(3)The expression of Caspase-9 in control group,H/R,H/R+LB,H/R+MB,H/R+HB group [(0.08 ± 0.01)VS(0.34 ± 0.01)VS(0.27 ± 0.01)VS(0.15 ±0.01)VS(0.12 ± 0.01),F=546.25,P<0.01].Comparison of expression levels of Caspase-9 in eachg group: compared with the control group,the expression of Caspase-9 in the H/R group increased,and the differences were statistically significant,the t value was ?,P value was less than 0.01.Compared with H/R group,the expression of Caspase-9 in H/R+LB,H/R+MB and H/R+HB groups were all decreased,and the differences were statistically significant,the t values were 12.12,29.50,and 67.00,respectively,P values were all less than 0.01.Caspase-9 expression levels were compared between H/R+LB,H/R+MB,and H/R+HB groups,F= 226.50,P <0.01,the difference was statistically significant: between H/R + LB and H/R + MB,t = 14.36,P <0.01,between H/R + MB and H/R + HB,t=3.02,P = 0.094,the difference was not statistically significant,between H/R+LB and H/R+HB,t=46.00,P<0.01.(4)XIAP expression level in control,H/R,H/R+LB,H/R+MB,H/R+HB group[(0.58±0.02)VS(0.14±0.02)VS(0.33±0.03)VS(0.42 ± 0.02)VS(0.45 ± 0.02),F = 220.40,P <0.01].Comparison of expression levels of XIAP in each group:compared with the control group,the expression of XIAP in the H/R group was all decreased,and the differences were statistically significant,the t value was 76.21,the P value was less than 0.01.Compared with H/R group,the expression of XIAP in H/R+LB,H/R+MB and H/R+HB groups were all increased,and the differences were statistically significant,the t values were-28.00,-83.00,and-91.00,respectively,the P values were all less than 0.01.XIAP expression levels were compared betweenH/R+LB,H/R+MB,and H/R+HB groups,F=25.23,P<0.01,the difference was statistically significant: between H/R+LB and H/R+MB,t=-15.59,P<0.01,between H/R+MB and H/R+HB,t=-8.00,P=0.17,between H/R+LB and H/R+HB,t=-13.23,P<0.01.Conclusion:In the PE trophoblast model,the expression of anti-apoptotic proteins Bcl-2 and XIAP decreased and the expression of apoptotic proteins caspase-3 and caspase-9increased.Baicalin can play a role in the treatment of PE by regulating apoptosis,increasing Bcl-2,XIAP,and reducing the expression of caspase-3 and caspase-9. |
PDF Full Text Request