| Objective:Pre-eclampsia(PE),a common late pregnancy idiopathic disease, which etiology of this disorder is not known. the liver is the most common damaged organ in addition to placenta in PE. Previous researches have shown that induced expression of XIAP/Bcl-2 and increased expression of Caspase-9 in trophoblast cells may be associated with pathogenesis of preeclampsia. The hepatocyte apoptosis has ignored in PE. Baicalin has strong activion in antibacterial, antiviral, antioxidant effects, anti-inflammatory, anti-allergic, anti-tumor.Therefore, the purpose of this study was to investigate the effect of different doses of baicalin on the expression of XIAP /Bcl-2/ Caspase-9 in Liver cells and the hepatocyte apoptosis rate of Preeclampsia model rat. Methods:The 75 pregnant rats were randomly divided into 2 groups: control group 15, preeclampsia(PE) group 60; secondary randomization of PE:blank control subgroup(BC) 15, low dose baicalin intervention(LB) group15, medium-dose baicalin intervention(MB)group15, high-dose baicalin intervention(HB)group15. Administration on Day 13 of gestation, CG group-[ NS, 125 mg /(kg ? d),s.c.], PE group- [L-NAME, 125 mg /(kg ? d),s.c.]; day 16, BC group plus saline [0.5ml /, s.c.], LB group, MB group, HB group plus the respctive dose baicalin( i.p.). Day 10,13,16,19 of gestation, measured rat tail artery pressure and 24 h urine protein and observed the reflection of baicalin in each group. On Day 21 of gestation, the rats were sacrificed under ether anesthesia. The Liver was removed and fixed carefully. By immunohistochemical methods to detect the expression of XIAP/ Bcl-2/ Caspase-9 protein was localized in pregnant rat liver tissue, process the image using SCANSCOBC and carry out semi-quantitative analysis through measuring gray value. TUNEL method was used to detect apoptosis of pregnant rats liver cells in each group. Data are analyzed t in t test, F test and LSD-t test by SPSS 17.0 software packet, different time points were compared using repeated measures analysis of variance, P <0.05 was considered statistically significant. Results:1.The comparison of the indexes on Day 10 of CG/PE group before the model:pregnancies(rats are primiparity),tail systolic blood pressure[(109.20±4.51)VS(107.80±5.93)mm Hg,t=1.01,P=0.324], diastolic blood pressure[(77.47±9.93)VS(78.75±6.80)mm Hg, t=0.59,P=0.557],24 h urine protein[(6.24±0.27)VS(6.11±0.54)mg/24 h,t=0.85,P=0.401] there is no significant difference between two groups(P>0.05);2.The comparison of the indexes on Day 16(after administration L-NAME 3 days) between control group and PE group have significant difference(P<0.05):systolic blood pressure[(115.20±4.90)VS(135.47±5.43)mm Hg, t=10.72,P<0.001], diastolic blood pressure[(77.93±7.82)VS(89.40±4.75)mm Hg, t=4.01,P<0.001],24 h urine protei n [(6.21±0.28)VS(7.52±0.58)mg/24 h, t=7.79, P<0.001];3.The comparison of the indexes of pregnant rats on day 19(after 4 days of diffirence dose baicalin intervention):(1).CG group /BC group/LB group/MB group/HB group: systolic blood pressure[(118.67±4.95)VS(144.60±5.32)VS(130.13±5.71)VS(122.60±3.91)VS(108.87±17.72) mm Hg, F=43.78, P<0.001]; diastolic blood pressure[(81.60±6.67)VS(90.80±6.42)VS(82.40±7.60)VS(81.33±5.50)VS(75.73±11.54)mm Hg, F=5.12, P<0.001]; 24 h urine protein[(6.24±0.26)VS(11.14±0.62)VS(6.91±0.34)VS(7.07±0.28)VS(7.31±1.22)mg/24 h,F=42.72,P<0.001];(2)(1)The comparison of systolic blood pressure of groups: compared with CG group, the pressure of BC group is significant increased t=7.80,P<0.001.compared with BC group, the pressure of LB/MB/HB group is significant increased t=4.35/ 6.62/10.75,P=<0.001.There is statistically significant difference among LB/MB and HB group(P<0.05), [(LB VS MB)mm Hg, t=2.27,P=0.027], [(MB VS HB)mm Hg, t=4.13,P<0.001], [(LB VS HB)mm Hg, t=6.40, P<0.001].But, there is no significant difference between CG and MB group t=1.18, P=0.241;(2)The comparison of diastolic blood pressure of groups: compared with CG group, the pressure of BC group is significant increased t=3.22,P=0.002, LB group have no signifcant difference with CG t=0.09, P=0.926.Compared with BC group, the pressure of LB/MB/HB group is significant increased t=2.94/3.31/5.27,P=0.004/0.001/0.001. LB/MB and HB group:[(LB VS MB)mm Hg, t=0.37,P=0.71],[(MB VS HB)mm Hg, t=1.96, P=0.054],[(LB VS HB)mm Hg, t=2.33,P=0.023];(3)The comparison of 24 h urine protein of groups: compared with CG group, 24 h urine protein of BC group is significant increased t=20.53,P<0.001.Compared with BC group, the pressure of LB/MB/HB group is significant increased t=17.69/17.05/16.04,P=0.001.There is no statistically significant difference among LB/MB and HB group,[(LB VS MB)mg/24 h, t=0.64,P=0.521],[(MB VS HB)mm Hg, t=1.00,P=0.321],[(LB VS HB)mm Hg, t=1.65, P=0.104].4.By immunohistochemical methods to detect expression of /XIAP/ Bcl-2/Caspase-9 in five groups:(1)The expression of XIAP of in liver cells comparison: compared with CG, XIAP grey value of BC group is significant reduced t=53.18, P<0.05. Compared with BC group, XIAP grey value of LB/MB/HB group is significant increased t=23.27/33.28/43.59,P=<0.001. There is statistically significant difference among LB,MB and HB group(P<0.05)[(LB VS MB), t=10.02,P<0.001],[(MB VS HB),t=10.30,P<0.001],[(LB VS HB), t=20.32, P<0.001].(2) The expression of Bcl-2 of liver cells comparison: compared with CG, Bcl-2 grey value of BC group is significant reduced t=22.31,P<0.001. compared with BC group, Bcl-2 grey value of LB/MB/HB group is significant increased t=4.33/7.69/12.61, P=<0.001. There is statistically significant difference among LB,MB and HB group(P<0.05)[(LB VS MB),t=3.36,P=0.001],[(MB VS HB),t=4.92,P<0.001],[(LB VS HB), t=8.28, P<0.001].(3)The expression of Caspase-9 of livers cells comparison: compared with control group,Caspase-9 grey value of BC group is significant increased(t=46.90,P<0.001). compared with BC group,Caspase-9 grey value of LB/MB/HB group is signif icant reduced t=14.00/34.32/25.71, P=<0.001.There is statistically significant differe nce among LB, MB and HB group(P<0.05), [(LB VS MB),t=20.31,P<0.001],[(MB VS HB), t=8.61, P<0.001], [(LB VS HB), t=11.70, P<0.001].5.The comparison of Hepatocyte apoptosis rate:compared with CG, the hepatocyte apoptosis rate is significant increased t=18.33, P<0.001. Compared with BC group, the hepatocyte apoptosis rate of LB/MB/HB group is significant reduced(t=7.25/10.81/10.80, P=<0.001). there is statistically significant difference among LB,MB and HB group(P<0.05)[(LB VS MB), t=3.56, P<0.001], [(MB VS HB),t=3.55,P<0.001]. There is no statistically significant difference between MB and HB group(P=1.000>0.05). Conclusion:The results of this study show that the intervention of baicalin can increase the expression of XIAP/Bcl-2, reduce the expression of Caspase-9,and decreased the rate of hepatocyte apoptosis of liver cells in a dose-dependent characteristics, proved baicalin potentially valuable treatment of pre-eclampsia, the safty and effective dose of the baicalin is a question. |
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