健脾补肾方对亚临床甲减孕鼠胎盘分泌功能及NF-κB/MMP-9通路的调控作用

Regulatory Effect of JianpiBushen Recipe on the Secretion Function and Expressions of NF-κB and MMP-9 in Placental Tissues of Pregnant Rats with Subclinical Hypothyroidism

    摘要
  • 摘要: 目的 通过研究健脾补肾方对亚临床甲减孕鼠胎盘分泌功能及核转录因子(NF-κB)和基质金属蛋白酶9(MMP-9)表达的调控作用，探讨健脾补肾方的保胎机制。方法 将90只Wistar雌性大鼠随机分成2组：正常对照组10只，模型组80只。建立亚临床甲减动物模型，达到完全甲减状态的模型大鼠中随机挑选10只为中药低剂量孕前给药组（Q），予皮下注射剂量为9.5μg/（kg·d）的L-T4，连续9d，第10天放免法检测大鼠血清TSH、TT4，筛选成功的亚临床甲减大鼠模型，将亚临床甲减大鼠以雌∶雄=2∶1的比例合笼。通过观察阴道栓，结合阴道涂片找精子，判断是否交配，并确定D0天，然后随机分为4组：①模型组即亚临床甲减模型组；②中药低剂量组；③中药高剂量组；④西药组。放免法测定怀孕第10天（D10）的孕鼠血清绒毛膜促性腺激素（HCG）、雌二醇（E2）、孕酮（P）；免疫组化、Western Blot法检测NF-κB和MMP-9在各组孕鼠胎盘组织中的表达。结果 与模型组比较，D10天，各给药组的血清P、HCG水平均有显著增高（P＜0.01），E2水平Q组差异最显著（P<0.01）和X组有显著的变化（P<0.05）；免疫组化及Western Blot结果均显示各给药组胎盘组织NF-κB及MMP-9水平均显著提高（P＜0.05）。结论 健脾补肾方可以提高NF-κB和MMP-9在亚临床甲减孕鼠胎盘组织中的表达，促进亚临床甲减孕鼠胎盘的分泌功能。

     

    Abstract: OBJECTIVE To study the effects of JianpiBushen recipe(JBR) on the secretion function and Nuclear Kappa B(NF-κB) and matrix metalloproteinase-9（MMP-9）in placental tissues of pregnant rats with subclinical hypothyroidism(SCH) and investigate the protective mechanism of JBR. METHODS Ninety female adult Wistar rats were randomly divided into 2 groups: normal control group(false thyroid surgery was carried out， n=10) and model control group(total thyroidectomy was carried out， n=80).The animal model of SCH was established by using＂total removal of the rats thyroid 30 days， levothyroxine was given to the rats by subcutaneous injection at 9.5μg/(kg·d)＂， All rats in the model with overt hypothyroidism were selected 10 as low dose Chinese medicine before pregnant group(levothyroxine was given to the rats by subcutaneous injection at 9.5μg/kg·d and low dose Chinese medicine， Q)， others were injected levothyroxine subcutaneously at 9.5μg/(kg·d) for 9 days， serum TSH and TT4 were detected by radioimmunoassay on the tenth day and screening of SCH rats， the ratio of male to female=2∶1 was used as a cage. Through the observation of vaginal suppository and vaginal smear to find sperm to determine whether the mating and determine D0 day. Then randomly divided into 4 groups: ①The control group is SCH group(C) ②Low dose Chinese medicine group(L) ③High dose Chinese medicine group(H) ④Western medicine group(X). Serum chorionic gonadotropin (HCG)， estradiol (E2) and progesterone (P) of the pregnant rats with SCH on the 10*day after conception (D10) in each group were detected by radioimmunoassay； The expressions of NF-κB and MMP-9 in placental tissues of pregnant rats with SCH on D10 in each group were detected by immunohistochemistry and Westernblottingassay. RESULTS Compared with the C group， on the tenth day， the serum levels of P and HCG in each treatment were all significantly higher (P<0.01)， the serum levels of E2 were significantly higher in the X group(P<0.05)， The results of immunohistochemistry and Western Blot showed that the expressions of NF-κB and MMP-9 in placental tissues in each treatment group were all significantly higher (P<0.05). CONCLUSION JBR can promote the secretion of placenta and increase the expressions of NF-κB and MMP-9 in the placenta of pregnant rats with SCH.

     

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