Effect of Huoxue Fang（活血方） on NF-κB Pathway and Matrix Metalloproteinases of Endometrium of Menstruation Bleeding Model Mice

CHEN Jincheng; LI Chunxiang; DING Fang; WU Beibei

doi:10.13288/j.11-2166/r.2015.11.018

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Effect of Huoxue Fang（活血方） on NF-κB Pathway and Matrix Metalloproteinases of Endometrium of Menstruation Bleeding Model Mice

更新时间：2021-11-17

- Effect of Huoxue Fang（活血方） on NF-κB Pathway and Matrix Metalloproteinases of Endometrium of Menstruation Bleeding Model Mice
- Journal of Traditional Chinese Medicine Vol. 56, Issue 11, Pages: 967-971(2015)
- 作者机构：
  
  1. 河北医科大学研究生院
  2. 河北中医学院
  3. 河北医科大学第三医院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2015.11.018
  CLC： R285.5
- Published：2015
- 稿件说明：
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[1]陈进成,李春香,丁芳,吴贝贝.活血方对月经出血模型小鼠子宫内膜NF-κB通路与金属基质蛋白酶的影响[J].中医杂志,2015,56(11):967-971.

CHEN Jincheng, LI Chunxiang, DING Fang, et al. Effect of Huoxue Fang（活血方） on NF-κB Pathway and Matrix Metalloproteinases of Endometrium of Menstruation Bleeding Model Mice[J]. Journal of Traditional Chinese Medicine2015, 56(11): 967-971.
[1]陈进成,李春香,丁芳,吴贝贝.活血方对月经出血模型小鼠子宫内膜NF-κB通路与金属基质蛋白酶的影响[J].中医杂志,2015,56(11):967-971. DOI： 10.13288/j.11-2166/r.2015.11.018.

CHEN Jincheng, LI Chunxiang, DING Fang, et al. Effect of Huoxue Fang（活血方） on NF-κB Pathway and Matrix Metalloproteinases of Endometrium of Menstruation Bleeding Model Mice[J]. Journal of Traditional Chinese Medicine2015, 56(11): 967-971. DOI： 10.13288/j.11-2166/r.2015.11.018.

摘要

目的探讨活血方对子宫内膜崩解的影响及可能作用机制。方法将130只C57BL/6J雌性小鼠随机分为假手术组10只、模型组60只、中药组60只。模型组与中药组均采用激素序贯处理复制小鼠月经出血模型

并对左侧宫腔诱导蜕膜化。从造模第9天中药组灌胃活血方20 g/（kg·d）

假手术组与模型组给予等量的生理盐水。在孕酮撤退后0、8、12、16、24、32 h分别于两组中选取一定只数小鼠进行阴道涂片检测

观察小鼠子宫内膜崩解脱落情况

HE染色观察双侧子宫角的病理变化并计算子宫系数

免疫组织化学法检测子宫组织MMP3、MMP9、P50、P65表达

Real-time PCR检测左侧子宫组织MMP3 mRNA、MMP9 mRNA的表达

Western blot方法检测左侧子宫组织MMP3、MMP9蛋白表达与P50、P65核蛋白表达。结果与假手术组比较

模型组小鼠子宫左侧有明显病理改变

从12 h出血小鼠比率增加

左侧子宫系数明显增加（P

0.05）;与模型组比较

中药组的出血时程明显缩短

在0、8、12 h左侧子宫系数增加、32 h降低（P

0.05）。与模型组比较

中药组左侧子宫组织MMP3 mRNA、MMP9 mRNA 0

2 h表达均明显上调（P

0.05）;MMP3及MMP9蛋白在0

2 h显著升高

32 h显著下调（P

0.05）;与模型组比较

中药组小鼠左侧子宫组织P50核蛋白在0、12 h显著上调

在8、16、24、32 h显著下调（P

0.05）;P65核蛋白在0、12、24 h显著上调

在32 h显著下调（P

0.05或P

0.01）。结论活血方可缩短子宫内膜崩解时程

其机制可能与调节崩解相关因子MMP3、MMP9的表达及NF-κB信号通路的快速激活与及时关闭相关。

Abstract

Objective To explore the effect of Huoxue Fang on disintegration of endometrium and possible mechanism. Methods Totally 130 C57 BL /6J female mice were divided into sham operation group（ n = 10）

model group（ n = 60） and herb group（ n = 60）. Model and herb group were given sequential hormone to replicate models of menstruation bleeding mice. Induce decidualization to left uterine cavity. From the ninth day of modeling

herb group were given Huoxue Fang 20 g /（ kg·d） by gavage. Sham operation group and model group were given equivalent normal saline. After progesterone withdrawals at 0

32 h

choose a certain amount of mice in both groups respectively to take vaginal secretion detection and observe disintegration and shedding of endometrium. After hematoxylin-eosin（ HE） staining

observe pathological change of bilateral uterine horn and calculate uterine coefficient. Detect the expression of MMP3、MMP9、P50 and P65 in uterine tissues adopting immunohistochemical method. Detect the expression of MMP3 mRNA and MMP9 mRNA with Real-time PCR. Detect the protein expression of MMP3 and MMP9 and the nuclear protein expression of P50 and P65 in uterine tissues using Western blot. Results Compared with sham operation group

left side of uterine of model group mice showed significant pathological change.From 12 h

the rate of bleeding mice increased; left uterine coefficient increased significantly（ P < 0. 05）. Compared with model group

bleeding time-histories of herb group decreased obviously; at 0

12 h

left uterine coefficient increased which decreased at 32 h（ P < 0. 05）. Compared with model group

the expression of MMP3 mRNA and MMP9 mRNA of left uterine tissues of herb group were up-regulated obviously during 0- 32 h（ P < 0. 05）. The expression of MMP3 and MMP9 protein increased during 0- 12 h and were down-regulated at 32 h（ P < 0. 05）.Compared with model group

nuclear protein P50 of left uterine tissues of herb group mice was up-regulated at 0 and12 h and was down-regulated at 8

32 h（ P < 0. 05）. Nuclear protein P65 was up-regulated at 0

24 h and was down-regulated at 32 h（ P < 0. 05 or P < 0. 01）. Conclusion Huoxue Fang can decrease disintegration time-histories of endometrium. The mechanism may relate to expression of MMP3 and MMP9 and rapid activation and timely closure of NF-κB pathway.

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