保产无忧散对孕晚期大鼠Cx-43基因表达及宫颈成熟相关指标的影响

Effects of Baochan Wuyou powder on Cx-43 gene expression and cervical maturation related indexes on late pregnancy in rats

目的:探讨保产无忧散对孕晚期大鼠Cx-43基因表达及宫颈成熟相关指标的影响。方法:取受孕成功的孕鼠30只,采用随机法将孕鼠分成模型对照组(孕鼠)、保产无忧散低剂量给药组(低剂量给药组)、保产无忧散高剂量给药组(高剂量给药组)三组,每组10只,另取11只未孕鼠作为正常对照组。保产无忧散煎煮滤过后浓缩为含生药1.17 g/ml和含生药0.585 g/ml。模型对照组、正常对照组均予以蒸馏水灌胃,保产无忧散低剂量给药组(0.585 g/ml)、高剂量给药组(1.17 g/ml)孕鼠于妊娠第16天开始给药,1次/d,均按1 ml/100 g体重灌胃给药。所有动物连续灌胃5 d。采用酶联免疫吸附剂法(ELISA)测定大鼠子宫体Cx-43含量及子宫前列腺素E 2(PEG 2)、宫颈白介素-8(IL-8)以及宫颈基质金属蛋白酶9(MMP-9)、基质金属蛋白酶组织抑制因子-1(TIMP-1),采用免疫组化法检测子宫体Cx-43阳性表达强度。结果:与正常对照组比较,模型对照组、低剂量给药组、高剂量给药组子宫Cx-43表达及子宫平滑肌中Cx-43阳性表达强度均显著升高(t=-8.593,-3.642;均P<0.01),与模型对照组比较,低剂量给药组Cx-43表达及子宫平滑肌中Cx-43阳性表达强度升高(t=-2.883,-1.950,P=0.005,0.033),高剂量给药组表达下降,差异无统计学意义(t=1.147、-0.653,P=0.133、0.261)。与正常对照组比较,模型对照组、低剂量组、高剂量组子宫组织中PEG_(2)、IL-8、MMP-9水平均显著升高(PEG_(2):t=-3.423、-6.004、-5.079,P=0.001、<0.001、<0.001;IL-8:t=-6.097、-7.995、-5.430,均P<0.001;MMP-9:t=-3.619、-6.172、-4.529,均P<0.001;TIMP-1:t=-2.522、-10.938、-10.938,P=0.011、<0.001、<0.001),与模型对照组比较,低剂量给药组子宫组织中PEG_(2)、IL-8、MMP-9、T1MP-1水平均升高(t=-2.157,-3.163,-2.671,-6.912;P=0.022,0.003,0.008,<0.001),高剂量给药组子宫组织中PEG_(2)、IL-8、MMP-9、TIMP-1水平均升高,但差异无统计学意义(t=-1.304、-0.567、-1.189、-1.610,P=0.104、0.289、0.125、0.062)。结论:低剂量保产无忧散通过提高孕晚期大鼠子宫组织中Cx-43表达及PEG_(2)、MMP-9和IL-8含量,参与并介导宫颈成熟。

Objective:To investigate the effects of Baochan Wuyou powder on Cx-43 gene expression and cervical maturation related indexes on late pregnancy in rats.Methods:30 pregnant rats were randomly divided into three groups:model control group(pregnant rats),low-dose administration group and high-dose administration group,with 10 rats in each group,and 11 non pregnant rats were taken as the normal control group.After decocting and filtering,Baochan Wuyou powder is concentrated into 1.17 g/ml and 0.585 g/ml of crude drugs.The model control group and the normal control group were given distilled water by gavage.The pregnant rats in the low dose group(0.585 g/ml)and the high dose group(1.17 g/ml)of Baochan Wuyou powder were administered by gavage at the weight of 1ml/100 g on the 16 th day of pregnancy.All animals were gavaged continuously for 5 days.The content of Cx-43 in rat uterine body,uterine prostaglandin E_(2)(PEG_(2)),cervical interleukin-8(IL-8),cervical matrix metalloproteinase-9(MMP-9)and tissue inhibitor of metalloproteinase-1(TIMP-1)were measured by enzyme-linked immunosorbent assay(ELISA).The positive expression intensity of Cx-43 in uterine body was detected by immunohistochemistry.Results:Compared with the normal control group,the expression of Cx-43 in uterus and the intensity of Cx-43 positive expression in uterine smooth muscle in model control group,low-dose group and high-dose group were significantly increased(t=-8.593,-3.642,all P<0.01).Compared with the model control group,the expression of Cx-43 and Cx-43 positive expression in uterine smooth muscle in low-dose group were significantly increased(t=-2.883,-1.950,P=0.005,0.033),The expression decreased in the high-dose group,and the difference was not statistically significant(t=1.147,-0.653,P=0.133,0.261).Compared with the normal control group,the levels of PEG 2,IL-8,MMP-9 and TIMP-1 in the uterine tissue of the model control group,low-dose group and high-dose group increased significantly on average(PEG_(2):t=-3.423,-6.004,-5.079,P=0.001,<0.001,<0.001;IL-8:t=-6.097,-7.995,-5.430,all P<0.001;MMP-9:t=-3.619,-6.172,-4.529,all P<0.001;TIMP-1:t=-2.522,-10.938,-10.938,P=0.011,<0.001).Compared with the model control group,the levels of PEG_(2),IL-8,MMP-9 and TIMP-1 were increased in the uterine tissue of the low-dose group(t=-2.157,-3.163,-2.671,P=0.022,0.003,0.008).The levels of PEG_(2),IL-8,MMP-9 and TIMP-1 were increased on average in uterine tissue of high-dose group,but the difference was not statistically significant(t=-1.304,-0.567,-1.189,P=0.104,0.289,0.125).Conclusion:Low dose Baochan Wuyou powder participates in and mediates cervical maturation by increasing the expression of Cx-43 and the contents of PEG_(2),MMP-9 and IL-8 in uterine tissue of rats in the third trimester of pregnancy.