慢病毒介导胰岛素基因增强子结合蛋白1对拉贝洛尔治疗妊娠期高血压孕鼠状态及子代心脏发育的影响
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摘要
目的研究慢病毒介导胰岛素基因增强子结合蛋白1(Islet-1)对拉贝洛尔治疗妊娠期高血压孕鼠状态及子代心脏发育的影响及其可能机制。方法按照体重将怀孕SD大鼠随机分为4组:正常妊娠组、空白对照组、阴性对照组和实验组,每组18只。除了正常妊娠组以外,其他3组大鼠经亚硝基左旋精氨酸诱导法建立妊娠期高血压模型。建模后、于妊娠第14天,阴性对照组注入LV慢病毒空载液100μL;实验组注入1. 0×108TU·m L~(-1) LV-Islet-1过表达重组慢病毒载体液100μL;于妊娠第16天,以上2组均给予拉贝洛尔溶液5mg·kg~(-1)·d~(-1)和硫酸镁注射液0. 1 g·kg~(-1)·d~(-1);空白对照组和正常妊娠组则给予等量0. 9%Na Cl。用逆转录-聚合酶链反应检测各组Islet-1 mRNA表达水平以评估慢病毒构建;用免疫印迹法检测各组给药前后心脏组织组蛋白H3乙酰化水平;监测孕鼠的收缩压。结果给药前,空白对照组、阴性对照组、正常妊娠组和实验组的孕鼠心脏组织中Islet-1 mRNA表达水平分别为0. 75±0. 03,0. 82±0. 02,1. 47±0. 13和3. 95±0. 21,空白对照组和阴性对照组比较,差异无统计学意义(P>0. 05)。给药前,这4组的乙酰化H3蛋白(光密度值)分别为(8. 4±1. 2)×10~(-2),(7. 8±1. 8)×10~(-2),(8. 1±1. 5)×10~(-2)和(8. 3±1. 9)×10~(-2);给药后,这4组的乙酰化H3蛋白水平分别为(7. 9±1. 3)×10~(-2),(7. 6±1. 6)×10~(-2),(8. 0±1. 4)×10~(-2)和(7. 9±1. 6)×10~(-2),给药前和后比较,这4组的差异均无统计学意义(均P> 0. 05)。给药前,阴性对照组和实验组的收缩压分别为(147. 45±8. 92),(136. 08±9. 15) mm Hg;给药后,这2组的收缩压分别为(125. 95±8. 06),(112. 35±7. 19) mm Hg,给药前和后比较,这2组的差异均有统计学意义(均P <0. 05);给药后实验组和阴性对照组比较,差异有统计学意义(P <0. 05)。结论过表达Islet-1可有效提高拉贝洛尔治疗妊娠期高血压的疗效,孕鼠和子代状况都得到明显改善,这可能与其参与组蛋白乙酰化相关。
Objective To study the effect of lentivirus mediated insulin gene enhancer binding protein 1( Islet-1) on the labetalol for treating cardiac development of the offspring of pregnant rats with pregnancy-induced hypertension. Methods Pregnant SD rats were randomly divided into four groups according to body weight: normal pregnancy group,blank control group,negative control group and experimental group,each group was 18 rats. In addition to the normal pregnancy group,the other three groups of rats were induced by nitroso-L-arginine to establish pregnancy-induced hypertension model. After modeling,on the 14 th day of pregnancy,the negative control group was injected with 100 μL of LV lentivirus no-load solution; the experimental group was injected with 1. 0 × 108 TU·m L~(-1) LV-Islet-1 overexpression recombinant lentivirus vector solution of 100 μL. On the 16 th day of pregnancy,the above two groups were given labellol solution( 5 mg·kg~(-1)·d~(-1)) and magnesium sulfate injection( 0. 1 g·kg~(-1)·d~(-1)),while blank control group and normal pregnancy group were given the same amount of 0. 9% NaCl. The expression of Islet-1 mRNA in each group was detected by reverse transcription polymerase chain reaction to evaluate the construction of lentivirus. The acetylation level of histone H3 in cardiac tissue before and after administration were detected by Western blotting. The systolic blood pressure( SBP) of pregnant rats were monitored. Results Before administration,the expression levels of Islet-1 mRNA in heart tissues of pregnant rat in blank control group,negative control group,normal pregnancy group and experimental group were 0. 75 ± 0. 03,0. 82 ± 0. 02,1. 47 ± 0. 13 and 3. 95 ± 0. 21,respectively; there was no significant difference between the blank control group and the negative control group( P > 0. 05). Before administration,acetylated H3 protein levels( OD) in the four groups were( 8. 4 ± 1. 2) × 10~(-2),( 7. 8 ± 1. 8) × 10~(-2),( 8. 1 ± 1. 5) × 10~(-2) and( 8. 3 ± 1. 9) ×10~(-2),respectively; after administration,acetylated H3 protein levels in the four groups were( 7. 9 ± 1. 3) × 10~(-2),( 7. 6 ± 1. 6) × 10~(-2),( 8. 0 ± 1. 4) × 10~(-2) and( 7. 9 ± 1. 6) × 10~(-2),respectively; there was no significant difference in the four groups before and after treatment( all P > 0. 05). Before administration,the SBP in negative control group and experimental group were( 147. 45 ± 8. 92),( 136. 08 ± 9. 15) mm Hg,respectively; after administration,the SBP in the two groups were( 125. 95 ± 8. 06),( 112. 35 ± 7. 19) mm Hg. There were significant differences in the two group before and after administration( P < 0. 05); there were a significant difference between the two groups after administration( P < 0. 05). Conclusion Overexpression of Islet-1 can effectively improve the efficacy of hypertension of pregnancy by labetalol,and the status of pregnant and offspring has been significantly improved,which may be related to its involvement in histone acetylation.
Objective To study the effect of lentivirus mediated insulin gene enhancer binding protein 1( Islet-1) on the labetalol for treating cardiac development of the offspring of pregnant rats with pregnancy-induced hypertension. Methods Pregnant SD rats were randomly divided into four groups according to body weight: normal pregnancy group,blank control group,negative control group and experimental group,each group was 18 rats. In addition to the normal pregnancy group,the other three groups of rats were induced by nitroso-L-arginine to establish pregnancy-induced hypertension model. After modeling,on the 14 th day of pregnancy,the negative control group was injected with 100 μL of LV lentivirus no-load solution; the experimental group was injected with 1. 0 × 108 TU·m L~(-1) LV-Islet-1 overexpression recombinant lentivirus vector solution of 100 μL. On the 16 th day of pregnancy,the above two groups were given labellol solution( 5 mg·kg~(-1)·d~(-1)) and magnesium sulfate injection( 0. 1 g·kg~(-1)·d~(-1)),while blank control group and normal pregnancy group were given the same amount of 0. 9% NaCl. The expression of Islet-1 mRNA in each group was detected by reverse transcription polymerase chain reaction to evaluate the construction of lentivirus. The acetylation level of histone H3 in cardiac tissue before and after administration were detected by Western blotting. The systolic blood pressure( SBP) of pregnant rats were monitored. Results Before administration,the expression levels of Islet-1 mRNA in heart tissues of pregnant rat in blank control group,negative control group,normal pregnancy group and experimental group were 0. 75 ± 0. 03,0. 82 ± 0. 02,1. 47 ± 0. 13 and 3. 95 ± 0. 21,respectively; there was no significant difference between the blank control group and the negative control group( P > 0. 05). Before administration,acetylated H3 protein levels( OD) in the four groups were( 8. 4 ± 1. 2) × 10~(-2),( 7. 8 ± 1. 8) × 10~(-2),( 8. 1 ± 1. 5) × 10~(-2) and( 8. 3 ± 1. 9) ×10~(-2),respectively; after administration,acetylated H3 protein levels in the four groups were( 7. 9 ± 1. 3) × 10~(-2),( 7. 6 ± 1. 6) × 10~(-2),( 8. 0 ± 1. 4) × 10~(-2) and( 7. 9 ± 1. 6) × 10~(-2),respectively; there was no significant difference in the four groups before and after treatment( all P > 0. 05). Before administration,the SBP in negative control group and experimental group were( 147. 45 ± 8. 92),( 136. 08 ± 9. 15) mm Hg,respectively; after administration,the SBP in the two groups were( 125. 95 ± 8. 06),( 112. 35 ± 7. 19) mm Hg. There were significant differences in the two group before and after administration( P < 0. 05); there were a significant difference between the two groups after administration( P < 0. 05). Conclusion Overexpression of Islet-1 can effectively improve the efficacy of hypertension of pregnancy by labetalol,and the status of pregnant and offspring has been significantly improved,which may be related to its involvement in histone acetylation.
引文
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[4]BROWN C,MCFARLANE-ANDERSON N,ALEXANDER-LINDOR,et al.The effects of S-nitrosoglutathione and S-nitroso-N-acetyl-D,L-penicillamine in a rat model of pre-eclampsia[J].JNat Sci Biol Med,2013,4(2):330-335.
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[6]杨春梅,许皓,易勤,等.Islet-1促C3H10T1/2细胞向心肌样细胞分化过程中的电生理特性[J].临床心血管病杂志,2015,31(10):1100-1105.
[7]余仲苏,孔娟娟,孙慧超,等.组蛋白H3高乙酰化对心脏相关转录因子Islet-1的调控作用[J].重庆医科大学学报,2012,37(6):502-506.
[2]IORDACHE F,CONSTANTINESCU A,ANDREI E,et al.Histone acetylation regulates the expression of Hox D9 transcription factor in endothelial progenitor cells[J].Rom J Morphol Embryol,2015,56(1):107-113.
[3]文睿婷,黄琳,于芝颖,等.甲基多巴和拉贝洛尔治疗妊娠期高收缩压疾病的系统评价[J].中国临床药理学杂志,2017,33(17):1710-1712.
[4]BROWN C,MCFARLANE-ANDERSON N,ALEXANDER-LINDOR,et al.The effects of S-nitrosoglutathione and S-nitroso-N-acetyl-D,L-penicillamine in a rat model of pre-eclampsia[J].JNat Sci Biol Med,2013,4(2):330-335.
[5]孔娟娟.Islet-1在心脏发育的乙酰化调控网络中的枢纽作用[D].重庆:重庆医科大学,2012.
[6]杨春梅,许皓,易勤,等.Islet-1促C3H10T1/2细胞向心肌样细胞分化过程中的电生理特性[J].临床心血管病杂志,2015,31(10):1100-1105.
[7]余仲苏,孔娟娟,孙慧超,等.组蛋白H3高乙酰化对心脏相关转录因子Islet-1的调控作用[J].重庆医科大学学报,2012,37(6):502-506.