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

张吟; 许小凤

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

张吟^1,,
许小凤^1,

1.
南京中医药大学第一临床医学院，江苏南京 210023
2.
南京中医药大学附属常州市中医医院，江苏常州 213003
3.
南京中医药大学附属苏州市中医医院，江苏苏州 215009

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出版历程
- 刊出日期: 2017-05-10

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

ZHANGYin^1
,,
XUXiao-feng^1
,

1.
The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing , 210023, China
2.
Changzhou Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, Changzhou , 213003, China
3.
Suzhou Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, Suzhou, 21500, China

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。
- 健脾补肾方 /
- 亚临床甲减孕鼠 /
- NF-κB /
- MMP-9
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.
- JianpiBushen recipe /
- pregnant rats with SCH /
- Nuclear Kappa B /
- matrix metalloproteinase-9