主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2019, Vol. 42 ›› Issue (9): 742-752.doi: 10.3969/j.issn.1006-2157.2019.09.008

• 中药药理 • 上一篇    下一篇

滋水清肝理冲饮对子宫内膜异位症小鼠应用GnRHa后所致神经递质紊乱的干预作用*

马小娜1, 谢苗1, 李冰冰2, 程发峰3, 王雪茜3, 郭亚楠1, 刘雁峰2#   

  1. 1 北京中医药大学第三附属医院 北京 100029;
    2 北京中医药大学东直门医院;
    3 北京中医药大学
  • 收稿日期:2019-03-04 出版日期:2019-09-30 发布日期:2019-09-27
  • 通讯作者: 刘雁峰,女,博士,教授,博士生导师,研究方向:中医药治疗女性不孕症相关疾病、女性生殖器官良性肿瘤,E-mail:liuyaf8888@sina.com
  • 作者简介:马小娜,女,博士,副主任医师,硕士生导师
  • 基金资助:
    国家自然科学基金资助项目(No.81273789),北京中医药大学自主选题项目(No.2015-JYB-JSMS133)

Effects of Zishui Qinggan Lichong Granule on neurotransmitter disorder in EMs mice induced by GnRHa*

Ma Xiaona1, Xie Miao1, Li Bingbing2, Gheng Fafeng3, Wang Xueqian3, Guo Yanan1, Liu Yanfeng2#   

  1. 1 The Third Affiliated Hospital of Beijing University of Chinese Medicine, Beijing 100029, China;
    2 Dongzhimen Hospital of Beijing University of Chinese Medicine, Beijing 100700, China;
    3 Beijing University of Chinese Medicine, Beijing 100029, China
  • Received:2019-03-04 Online:2019-09-30 Published:2019-09-27
  • Contact: Prof.Liu Yanfeng, Ph.D., Doctoral Supervisor. Dongzhimen Hospital of Beijing University of Chinese Medicine. No. 5 Haiyuncang Road, Dongcheng District, Beijing 100029. E-mail:liuyaf8888@sina.com
  • Supported by:
    National Natural Science Foundation of China (No. 81273789)

摘要: 目的 研究滋水清肝理冲饮对子宫内膜异位症(EMs)小鼠应用促性腺激素释放激素(GnRHa)后脑组织单胺类神经递质的影响;并通过对其在位、异位内膜组织中免疫因子的检测验证其应用的安全性。方法 选用BALB/c小鼠,随机分为正常组、模型组、GnRHa组、滋水清肝理冲饮高剂量组、滋水清肝理冲饮中剂量组、滋水清肝理冲饮低剂量组。除正常组外,其他各组采用腹腔注射的方法造模,把捐赠鼠的子宫内膜碎片注入小鼠腹腔,建立子宫内膜异位症小鼠模型。造模后除正常组和模型组外,其余各组在造模21 d后开始给予GnRHa(1.4 mg/kg)皮下注射1次,在给予GnRHa皮下注射日当天,滋水清肝理冲饮高、中、低剂量组同时开始给予滋水清肝理冲饮配方颗粒灌胃(48、24、12 g/kg),每天1次,连续给药30 d;正常组、模型组和GnRHa组给予相同体积的生理盐水灌胃。给药结束后,采用ELISA法检测各组小鼠脑组织5-羟色胺(5-HT)、去甲肾上腺素(NE)、多巴胺(DA)的含量,采用免疫组化法检测各组小鼠肿瘤坏死因子-α(TNF-α)白细胞介素-1(IL-1)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶组织抑制因子2(TIMP-2)的表达。结果 应用GnRHa后脑组织5-HT、NE、DA含量较模型组降低;滋水清肝理冲饮低、中、高剂量干预后脑组织5-HT含量均升高(均P<0.05),滋水清肝理冲饮中、高剂量组脑组织NE、DA含量升高(均P<0.05)。与疾病模型组相比,应用GnRHa后在位、异位内膜组织TNF-α、IL-1、MMP-2表达降低,TIMP-2表达升高;滋水清肝理冲饮各组干预后未出现TNF-α、IL-1、MMP-2表达升高,TIMP-2表达降低的情况。结论 GnRHa可导致小鼠脑组织单胺类神经递质5-HT、NE、DA含量下降,而滋水清肝理冲饮可有效上调小鼠脑组织单胺类神经递质水平,调节GnRHa所致围绝经期相关神经递质的紊乱。并且,改善GnRHa所致副反应的同时,不导致子宫内膜异位症免疫功能紊乱,安全性很好。

关键词: 滋水清肝理冲饮, 子宫内膜异位症, 促性腺激素释放激素, 单胺类神经递质, 免疫因子

Abstract: Objective To observe the effects of Zishui Qinggan Lichong Granule (ZQLG) (Water-nourishing, Liver-heat-clearing and Chong mai-regulating Granule) on monoamine neurotransmitter in brain tissue of mice with endometriosis (EMs) induced by gonadotropin releasing hormone agonist (GnRHa) and to verify its safety by monitoring the immunological factors in eutopic and ectopic endometrial tissues. Methods BALB/c mice were randomly divided into normal group, model group, GnRHa group, high-dose ZQLG group, mid-dose ZQLG group and low-dose ZQLG group. Except for the normal group, endometrial fragments of donated mice were injected intraperitoneally into the abdominal cavity of mice in other groups to establish the mouse model of endometriosis. Except for the normal group and the model group, GnRHa (1.4 mg/kg) was injected once subcutaneously to the other groups since the 21st day after the model establishment. From the day of subcutaneous injection of GnRHa, the high-, mid- and low-dose ZQLG groups began to receive Zishui Qinggan Lichong Granule by gavage [48, 24, 12 g/kg] for 30 days, and the normal group, model group and GnRHa group were given the same volume of saline by gavage. After the intervention, the content of 5-HT, NE and DA in brain tissue of mice in each group was detected by ELISA, and the expression of TNF-α, IL-1, MMP-2 and TIMP-2 was detected by immunohistochemistry. Results The contents of 5-HT, NE and DA in brain tissue after GnRHa injection were lower than those in the model group. The contents of 5-HT in brain tissue increased after ZQLG intervention in the low-, mid- and high-dose ZQLG groups(P<0.05). The contents of NE and DA in brain tissue increased in the mid- and high-dose ZQLG groups(P<0.05). Compared with model group, the expressions of TNF-α, IL-1 and MMP-2 in eutopic and ectopic endometrial tissues decreased and expression of TIMP-2 increased after GnRHa injection. There was no increase in TNF-α, IL-1, MMP-2 expressions or decrease in TIMP-2 expressions in any ZQLG group after intervention. Conclusion GnRHat led to a decline in the contents of such monoamine neurotransmitters as 5-HT, NE, and DA in mice brain tissue. However, Zishui Qinggan Lichong Granule can effectively increase the levels of such monoamine neurotransmitters in mice brain tissue and relieve perimenopausal neurotransmitter disorders caused by GnRHa. At the same time, it does not cause immune dysfunction in endometriosisits, delivering satisfying safety.

Key words: Zishui Qinggan Lichong Granule, endometriosis, GnRHa, monoamine neurotransmitter, immunological factor

中图分类号: 

  • R285.5