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 ›› 2020, Vol. 43 ›› Issue (5): 408-413.doi: 10.3969/j.issn.1006-2157.2020.05.010

• 中药药理 • 上一篇    下一篇

脑泰方Ⅱ号含药血清对LPS诱导的小胶质细胞极化的调节作用*

张秀丽1,2, 雷昌1,2, 刘洋3, 葛金文3#, 孟盼1,2, 张君宇1, 任永镇1, 资冬1, 朱伟1   

  1. 1.湖南中医药大学科技创新中心 湖南 410208;
    2.湖南省中药粉体与创新药物省部共建国家重点实验室培育基地;
    3.湖南中医药大学心脑疾病中西医结合防治湖南省重点实验室
  • 收稿日期:2019-11-26 发布日期:2020-06-08
  • 通讯作者: #葛金文,男,教授,博士生导师,主要研究方向:心脑血管疾病的中西医结合防治,E-mail:40831556@qq.com
  • 作者简介:张秀丽,女,博士,副研究员
  • 基金资助:
    *国家自然科学青年基金项目(No.81603608、8174174),教育部博士后面上项目(No.2018M630905),湖南省自然科学青年基金项目(No.2019JJ50431),中医内科重大疾病防治研究与转化教育部重点实验室开放基金(No.ZYNK201708),湖南省大学生研究性学习和创新性实验计划项目(No.1021-0001017161),湖南中医药大学中医学一流学科开放基金(No.4901-020000200221)

Effect of medicated serum of NaotaifangⅡ on microglia polarization induced by LPS*

Zhang Xiuli1,2, Lei Chang1,2, Liu Yang3, Ge Jinwen3#, Meng Pan1,2, Zhang Junyu1, Ren Yongzhen1, Zi Dong1, Zhu Wei1   

  1. 1. Technology Innovation Center, Hunan University of Chinese Medicine, Hunan 410208, China;
    2. State Key Laboratory Breeding Base of TCM Powder and Innovative Drugs Co-founded by Hunan Province and the Ministry of Science and Technology, Hunan 410208, China;
    3. Key Lab of Hunan Province for Prevention and Treatment of Cardio-cerebral Diseases with Integrated Traditional Chinese and Western Medicine, Hunan University of Chinese Medicine, Hunan 410208, China
  • Received:2019-11-26 Published:2020-06-08
  • Contact: Prof. Ge Jinwen, Doctoral Supervisor. Key Lab of Hunan Province for Prevention and Treatment of Cardio-cerebral Diseases with Integrated Traditional Chinese and Western Medicine, Hunan University of Chinese Medicine, No. 300, Xueshi Road, Hanpu Science and Education Park, Yuelu District, Changsha 410208, China. E-mail: 40831556@qq.com
  • Supported by:
    National Natural Science Foundation for Young Scientists of China(No.81603608,8174174); Chinese Postdoctoral Science Foundation (No.2018M630905); Natural Science Project of Hunan Province (No.2099JJ50431)Ethical review: Committee of Ethical Review on Experimental Animals of First Affiliated Hospital of Hunan University of Chinese Medicine (No.ZYFY20180227)

摘要: 目的 探明脑泰方Ⅱ号含药血清对LPS诱导的HAPI小胶质细胞形态、M1/M2型极化标记分子及炎性因子的影响。方法 建立LPS诱导的HAPI细胞炎症模型,将培养的细胞分为:空白组、LPS模型组、脑泰方Ⅱ号组、米诺环素组。高内涵观察细胞形态与数量的变化,Real-time PCR检测M1型标志分子MHCⅡ、iNOS、MCP-1、CD11b,M2型标志分子Arg1、Mrc1、Ym1,促炎性因子IL-1β、IL-6、IL-12、TNF-α,抗炎性因子IL-4、IL-10、TGF-β mRNA水平。结果 与正常组比较,LPS模型组细胞胞体变大,并呈多极化或阿米巴型,突触变粗变短;其M1型标记分子与促炎性因子的mRNA水平显著升高(P<0.01);而M2型标记分子Mrc1与抗炎性因子IL-4、TGF-β的mRNA水平显著降低(P<0.01)。经脑泰方Ⅱ号干预后,细胞胞体胀大部分恢复,突触变长,且细胞数量增多;M1型标记分子MHCⅡ、iNOS、CD11b与促炎性因子的mRNA水平显著降低(P<0.01), M2型标记分子与抗炎性因子的mRNA水平显著升高(P<0.01)。与米诺环素组比,脑泰方Ⅱ号对M1型标志分子mRNA水平的降低作用无明显优势(P>0.05),对促炎性因子IL-6、IL-12、TNF-α的mRNA水平的降低作用虽优于米诺环素组(P<0.01),但对IL-1β的调节作用与米诺环素组相比无显著差异(P>0.05);其对M2型标志分子的Mrc1、Ym1及抗炎性因子IL-4、TGF-β mRNA水平的提升作用均明显优于米诺环素组(P<0.01或P<0.05)。结论 脑泰方Ⅱ号可通过对细胞形态、M1/M2型标记分子与炎性因子的调节作用抑制LPS诱导的HAPI细胞M1型极化,并促进M2型极化。脑泰方Ⅱ号对M2型标志分子及抗炎性因子的调节作用较米诺环素具有一定优势。

关键词: 脑泰方Ⅱ号, 小胶质细胞极化, M1型标记分子, M2型标记分子, 促炎性因子, 抗炎性因子

Abstract: Objective To investigate the influence of medicated serum of Naotaifang Ⅱ on the morphology, M1/M2 polarization markers and inflammatory factors of HAPI cells induced by LPS. Methods The inflammation model of HAPI cell was induced by LPS. The cultured cells were divided into: blank group, model group, Naotaifang Ⅱ group, and minocycline group. Morphology and quantity of microglia were observed by High Content Imaging System. mRNA expression of M1-type markers (MCH II, iNOS, MCP-1, CD11b) and M2-type markers (Arg1, Mrc1, Ym1), pro-inflammatory factors (IL-1β, IL-6, IL-12, TNF-α) and anti-inflammatory factors (IL-4, IL-10, TNF-β) was detected by qRT-PCR. Results Compared with the blank group, in the model group, microglia cell body became larger while showing multipolarization or amoeba type with coarser and shorter synapse; the mRNA level of M1 markers and pro-inflammatory increased significantly (P< 0.01); moreover, the expression of M2 marker Mrc1 and anti-inflammatory factors IL-4 and TNF-β decreased dramatically (P<0.01). Compared with the model group, in NaotaifangⅡgroup, the cell swelling recovered, the synapses became longer, and the microglia increased. Besides, in NaotaifangⅡgroup, the mRNA expression of M1 markers MHCⅡ, iNOS and CD11b and the pro-inflammatory factors were significantly decreased (P<0.01), and the mRNA expression of M2 markers and anti-inflammatory factors were significantly increased (P<0.01). Compared with the minocycline group, the Naotaifang Ⅱ group showed no significant advantage in reducing the expression of M1 markers (P>0.05), while better effect on lowering the expression of pro-inflammatory factors IL-6, IL-12 and TNF-α (P<0.01) except IL-β. Besides, NaotaifangⅡcould better increase mRNA expression of M2 markers Mrc1, Ym1 and anti-inflammatory factors IL-4, TGF-β compared with the minocycline group. Conclusion Naotaifang seems to inhibit M1 polarization, promote M2 polarization by regulating cell morphology, M1/M2 markers and inflammatory factors. It could possibly enhance M2 markers and anti-inflammatory factors better than minocycline.

Key words: Naotaifang Ⅱ, microglia polarization, M1 markers, M2 markers, pro-inflammatory factors, anti-inflammatory factors

中图分类号: 

  • R285.5