六味地黄丸联合胰岛素对妊娠糖尿病大鼠血清皮质酮及胎盘11β-羟基类固醇脱氢酶表达的影响

Effects of Six-ingredient Rehmannia Pill Combined with Insulin on Expression of Serum Corticosterone and Placental 11β-hydroxysteroid Dehydrogenase in GDM Rats

目的探讨六味地黄丸联合胰岛素对妊娠糖尿病(GDM)大鼠血清皮质酮及胎盘11β-羟基类固醇脱氢酶(11β-HSDs)表达的影响。方法将受孕后的育龄雌性大鼠随机分为5组(每组8只):正常妊娠组(A组)、GDM组(B组)、胰岛素组(C组)、六味地黄丸组(D组)、六味地黄丸联合胰岛素组(E组)。受孕当日,B、C、D、E组均制备GDM模型。确定造模成功后,从孕第4天开始实施用药干预,C组单用胰岛素20U·kg^(-1)皮下注射,D组单用六味地黄丸10g·kg^(-1)灌胃,E组给予胰岛素与六味地黄丸联合用药(胰岛素20U·kg^(-1)皮下注射,六味地黄丸10g·kg^(-1)灌胃),A、B组用等体积蒸馏水代替。共用药14d。分别在孕前、用药前、末次给药禁食12h后取血,检测血糖、皮质酮的含量。分别用Western blot法、实时PCR法检测胎盘组织中11β-HSDs蛋白、11β-HSDs mRNA表达水平。结果用药前B、C、D、E组血糖、皮质酮水平均明显高于A组,差异有统计学意义(P<0.01),B、C、D、E组间血糖、皮质酮水平差异无统计学意义(P>0.05);用药后,C、D、E组血糖、皮质醇水平均明显低于B组(P<0.05或P<0.01),E组血糖、皮质醇水平明显低于C组及D组(P<0.05或P<0.01)。与A组比较,B、C、D组11β-HSD1mRNA、11β-HSD1蛋白表达明显升高,11β-HSD2mRNA表达明显降低,差异均有统计学意义(P<0.05);与B组比较,C、D、E组11β-HSD1mRNA、11β-HSD1蛋白表达明显降低,11β-HSD2mRNA表达明显升高,差异有统计学意义(P<0.05);E组分别与C、D组比较,11β-HSD1 mRNA表达均明显降低,11β-HSD2 mRNA表达均明显升高(P<0.01或P<0.05);5组间11β-HSD2蛋白表达差异无统计学意义(P>0.05)。结论六味地黄丸联合胰岛素用于GDM大鼠的治疗中,能明显降低GDM大鼠血糖水平。其可能是通过调节胎盘11β-HSDs mRNA表达,平衡皮质酮,抑制胰岛素抵抗,发挥降血糖作用。

Objective To investigate the effect of Six-ingredient Rehmannia Pill combined with insulin on the expression of 11β-hydroxysteroid dehydrogenase（11β-HSDs）in the placenta of gestational diabetes mellitus（GDM）rats.Methods Female rats of childbearing age（SD）were randomly divided into 5 groups（8 rats each）：normal pregnancy group（group A）,GDM group（group B）,insulin group（group C）,Six-ingredient Rehmannia Pill group（group D）,Six-ingredient Rehmannia Pill Combined with insulin group（group E）.GDM models were established in groups B,C,D and E on the day of conception.From the 4 th day of pregnancy,drug intervention was started.Group C was treated with insulin 20 U·kg-1 ih alone,group D with Six-ingredient Rehmannia Pill 10 g·kg-1 ig alone,while group E was given insulin combined with Six-ingredient Rehmannia Pill（insulin 20 U·kg-1,ih,Six-ingredient Rehmannia Pill 10 g·kg-1,ig）.The medication lasted for 14 days.Blood samples were taken before pregnancy.Before medication,at the last dose,and 12 hafter fasting,blood glucose and corticosterone levels were measured.The expression levels of 11β-HSDs protein and 11β-HSDs mRNA in placenta tissues were detected with Western blot and real-time PCR respectively.Results Before treatment,blood glucose and corticosterone levels in groups B,C,D and E were significantly higher than that in group A,and the difference was statistically significant（P 〈0.01）.There was no significant difference in blood glucose and corticosterone levels between groups B,C,D and E（P 〉0.05）.After medication,the levels of blood glucose and cortisol in groups C,D,and E were significantly lower than that in group B（P 〈0.05 or P 〈0.01）.The levels of blood glucose and cortisol in group E were significantly lower than those in group C and group D（P 〈0.05 or P〈 0.01）.Compared with group A,the expressions of11β-HSD1 mRNA and 11β-HSD 1 protein in the other four groups were significantly increased,while the expression of 11β-HSD2 mRNA decreased significantly,and the difference was statistically significant（P 〈0.05）.Compared with group B,the expressions of 11β-HSD1 mRNA and 11β-HSD1 protein in groups C,D,and E were significantly lower,but the expression of 11β-HSD2 mRNA increased significantly,and the difference was statistically significant（P 〈0.05）.There was no significant difference in the expression of 11β-HSD2 protein between the five groups（P〉 0.05）.Conclusion The Six-ingredient Rehmannia Pill combined with insulin,when used for treating GDM in rats,can obviously lower the blood glucose level of GDM rats,possibly by regulating the placental expression of 11β-HSDs mRNA,balancing corticosterone,inhibiting insulin resistance,and exerting hypoglycemic effect.