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-βmRNA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-βmRNA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。
Objective To investigate the influence of medicated serum of NaotaifangⅡon 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,NaotaifangⅡgroup,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,CD11 b)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-βdecreased 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 CD11 b 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 NaotaifangⅡgroup 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-βcompared 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
作者
张秀丽
雷昌
刘洋
葛金文
孟盼
张君宇
任永镇
资冬
朱伟
Zhang Xiuli;Lei Chang;Liu Yang;Ge Jinwen;Meng Pan;Zhang Junyu;Ren Yongzhen;ZiDong;Zhu Wei(Technology Innovation Center,Hunan University of Chinese Medicine,Hunan 410208,China;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;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)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2020年第5期408-413,共6页
Journal of Beijing University of Traditional Chinese Medicine
基金
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)
