疏肝健脾方对产后缺乳肝郁脾虚证模型大鼠JAK2/STAT5A信号通路的影响

Effects of Shugan Jianpi Recipe(疏肝健脾方) on JAK2/STAT5A Signaling Pathway in Model Rats of Postpartum Hypogalactia With Liver Depression and Spleen Deficiency Syndrome

目的探讨疏肝健脾方治疗产后缺乳肝郁脾虚证的可能作用机制。方法 60只Wistar临产期孕鼠随机分为模型组、空白对照组、西药对照组和疏肝健脾方低、中、高剂量组,每组10只。除空白对照组外,其余各组大鼠产后采用甲磺酸溴隐亭溶液+慢性束缚法复制大鼠产后缺乳肝郁脾虚病证模型。造模后西药对照组予甲氧氯普胺片溶液3. 15 mg/(kg·d)灌胃,疏肝健脾方低、中、高剂量组分别给予浓度为1. 109、2. 218、4. 435 g/ml疏肝健脾方溶液每只5 ml灌胃,模型组、空白对照组予等体积蒸馏水灌胃,各组均每日早、晚各灌胃1次,连续7天。采用RT-PCR法检测乳腺组织中酪氨酸激酶2 (JAK2)、信号传导及转录激活因子5A (STAT5A)、催乳素受体(PRLR)和β-酪蛋白(β-casein) mRNA表达,采用Western blot法检测乳腺组织中JAK2、STAT5A、PRLR和β-casein蛋白表达。结果与空白对照组比较,模型组、西药组和疏肝健脾方低、高剂量组大鼠乳腺组织JAK2、STAT5A、PRLR及β-casein mRNA及蛋白水平显著降低(P <0. 05),而疏肝健脾方中剂量组差异无统计学意义(P> 0. 05)。与模型组比较,西药对照组和疏肝健脾方各剂量组大鼠乳腺组织JAK2、STAT5A、PRLR及β-casein mRNA及蛋白水平均升高,且疏肝健脾方中剂量组明显高于其余给药组(P <0. 05)。结论疏肝健脾方治疗产后缺乳肝郁脾虚证的机制可能与其促进催乳素与其受体结合,激活JAK2/STAT5A信号转导通路,并上调β-casein基因表达相关。

Objective To explore the possible mechanism of Shugan Jianpi Recipe(疏肝健脾方)in the treatment of postpartum hypogalactia of liver depression and spleen deficiency syndrome.Methods A total of 60 Wistar pregnant rats with due date were randomly divided into model group,blank control group,western medicine control group and Shugan Jianpi Recipe low,medium and high dose groups,with 10 rats in each group.Except the blank control group,the rats were established the model of postpartum hypogalactia of liver depression and spleen deficiency syndrome using bromocriptine mesylate+chronic restraint method.After modeling,the western medicine control group was given 3.15 mg/(kg·d)of metoclopramide tablet solution,and the Shugan Jianpi Recipe low,medium and high dose groups were intragastrically administered with 1.109,2.218,4.435 g/ml of Shugan Jianpi Recipe solution respectively,5 ml per rat.The model group and the blank control group were intragastrically administered with equal volume of distilled water.Each group was intragastrically administered once in the morning and evening for 7 days.Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of tyrosine kinase 2(JAK2),signaling and transcriptional activator 5 A(STAT5A),prolactin receptor(PRLR)andβ-casein mRNA in breast tissue.The expression of JAK2,STAT5A,PRLR andβ-casein protein in breast tissue was detected by Western blot.Results Compared with the blank control group,the levels of JAK2,STAT5A,PRLR andβ-casein mRNA and protein in the mammary gland in the model group,the western medicine group and the Shugan Jianpi Recipe low and high dose group were significantly decreased(P<0.05),while there was no significant difference in the Shugan Jianpi Recipe middle dose group(P>0.05).Compared with the model group,the levels of JAK2,STAT5A,PRLR andβ-casein mRNA and protein in the mammary gland of rats in the western medicine control group and all Shugan Jianpi Recipe groups were increased,and the middle dose group was significantly higher than the other drug groups(P<0.05).Conclusion The mechanism of Shugan Jianpi Recipe in the treatment of postpartum hypogalactia of liver depression and spleen deficiency syndrome may be related to the promotion of prolactin binding to its receptor,activation of JAK2/STAT5A signal transduction pathway,and up-regulation ofβ-casein gene expression.