摘要
目的建立免疫性卵巢早衰(POF)动物模型,探讨模型小鼠POF的免疫调节机制,为临床研究POF的免疫机制提供实验依据。方法选取B6AF1雌鼠40只,分为空白组(20只)与模型组(20只)。模型组小鼠用透明带3(ZP3)的多肽片段(pZP3)为免疫原免疫小鼠建立自身免疫性POF小鼠模型。比较两组小鼠一般情况,以判断pZP3对模型小鼠状态的影响。14 d后两组小鼠均取脏器称重,计算脏器指数。两侧卵巢组织病理切片分别进行HE染色和免疫荧光染色,检测卵巢组织形态、淋巴细胞浸润和透明带抗体分布的情况。采用实时定量PCR检测叉状/翼状螺旋转录因子(Foxp3)及维甲酸相关孤核受体γt(ROR-γt)mRNA表达情况。结果经pZP3免疫的B6AF1小鼠一般情况较差,容易出现烦躁和愤怒等现象。与空白组相比,模型组小鼠脾脏指数升高(P<0.01),胸腺指数和卵巢指数均下降(P均<0.05),子宫指数有所下降,但无统计学差异(P>0.05);空白组卵巢组织中未见炎性细胞浸润及透明带抗体荧光,模型组小鼠卵巢组织中出现了不同程度的炎症细胞浸润,且有强度不等的透明带抗体荧光的显现。实时定量PCR结果显示,与空白组相比,模型组小鼠脾脏中Foxp3 mRNA相对表达水平明显下降(P<0.01),ROR-γt mRNA相对表达水平明显上升(P<0.05)。结论采用pZP3免疫B6AF1雌性小鼠能够成功建立免疫性POF模型,其中调节性T细胞(Treg细胞)和辅助性T细胞(Th)17中的特异性转录因子Foxp3和ROR-γt表达水平出现失衡,这有助于研究卵巢早衰的免疫调节机制。
Objective To establish animal model of immune premature ovarian failure （POF） and explore the immune regulatory mechanism of mice POF in to provide experimental basis for clinical research of immune POF. Methods Forty female B6AF1 mice were selected and divided into blank group and model group （n = 20 each）. Using mice zona pellueida 3 （ZP3） polypeptide fragment （pZP3） as immunogen, the autoimmune POF mouse model was established by immunizing mice with pZP3 in model group. General conditions of mice in two groups were compared to judge the influence of pZP3 on the state of model mice. After 14 days, mice organs were taken for weighing and calculating organ indexes in two groups. HE staining and immunofluoreseence staining were respectively performed on bilateral ovary pathological sections to detect ovarian tissue morphology,lymphocyte infiltration and distribution of zona pellucida antibody. Real-time fluorescent quantitative PCR （qRT-PCR） method was used to detect the expressions of forkhead or winged helix transcription 3 （FOXP3） mRNA and retinoid-related orphan nuclear receptor-yt （ROR-yt）mRNA. Results The general situation of B6AF1 mice immu- nized with pzp3 was poor, and the mice were prone to irritability, anger, etc. Compared with blank group, in the mode] group, spleen index increased （ P 〈0.01） ; thymus index and ovarian index decreased （ all P 〈0. 01 ） ; uterus indexs were similar between two groups （P 〉0. 05 ） , though it was lower somewhat in model group than that in blank group. No inflhra-tion of inflammatory cells in ovarian tissues and fluorescence of zona pellucida antibody were observed in blank group. There were inflammatory cell infiltration of different degrees in ovarian tissues and fluorescence of zona pellucida antibody with different intensity in model group. The results of qRT-PCR showed that compared with blank group, in model group, the relative expression level of Foxp3 mRNA in spleen significantly decreased （P 〈0. 01 ） ;the relativ
作者
王佩娟
陈思
卢燕
WANG Pei-juan CHEN Si LU Yan(Department of Gynecology and Obstetrics, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine , Nanjing University of Chinese Medicine （Jiangsu Province Traditional Chinese Medicine Research Institute）, Nanjing, Jiangsu 210028, China)
出处
《中国临床研究》
CAS
2017年第3期298-301,共4页
Chinese Journal of Clinical Research
基金
国家自然科学基金青年项目(81303136)
江苏省自然科学基金面上项目(BK20151606)
江苏省中医药局科技项目(LZ13088)
关键词
卵巢功能减退
卵巢早衰
动物模型
免疫调节
Hypoovarianism
Premature ovarian failure
Animal model
Immune regulation
