胸腺切除小鼠自身免疫性卵巢疾病发生发展的时相性变化

Temporal changes in the development of autoimmune ovarian diseases in thymectomized mice

目的通过建立小鼠自身免疫性卵巢疾病(autoimmune ovarian disease,AOD)模型,探索AOD中Treg、B淋巴细胞亚群的时相性变化,寻找干预疾病的窗口期,为人类AOD的早诊早治提供线索。方法建立出生第3天胸腺摘除Balb/c模型鼠(thymectomize on day3,D3tx),术后连续动态观察小鼠血清性激素(E2、FSH、LH)、炎症细胞因子(TGF-β、IL-4、IFN-γ、IL-21)、脾脏和盆腔淋巴结中Treg、B淋巴细胞亚群及卵巢组织学改变。结果 D3tx鼠术后第5周血清FSH、LH升高,伴E2下降;卵巢组织中出现较明显的炎细胞浸润及卵泡闭锁。从术后5周开始血清中炎症细胞因子TGF-β和IL-4降低,而IFN-γ和IL-21升高,持续到10周;D3tx组小鼠脾脏和腹股沟淋巴结,腹主动脉旁淋巴结中的Treg细胞在术后1~3周轻微升高,随后逐渐下降,5周后细胞数明显低于对照组(P<0.05);活化B细胞(CD138^-IgD^+)自术后1周起升高,并维持到10周(P<0.05);浆细胞(CD138^+CD19^-IgD^-)则从术后5周起升高,维持到10周(P<0.05)。结论机体免疫功能的时相性改变与AOD的发生发展密切相关,从免疫功能损害发生到卵巢组织学改变存在较明确的窗口期(即术后的1~4周),提示在早期干预和改善免疫功能状况可能改善自身免疫性卵巢疾病的转归。

This study was performed to explore the temporal changes of Treg and B lymphocyte subsets in a mouse model of autoimmune ovarian disease(AOD), with an aim of looking for a window of interventional diseases and providing clues for early diagnosis and treatment of human AOD. The thymus-extracted Balb/c model mice(thymectomized on day 3, D3tx) were established on the 3^rd day of birth. The serum levels of sex hormones(E2,FSH, LH) and inflammatory cytokines(TGF-β, IL-4, IFN-γ, IL-21) were observed continuously after operation;the lymphocyte subsets in spleen and pelvic lymph nodes, and the histological changes of ovary were observed. D3tx mice demonstrated higher serum FSH and LH levels, lower E2 level, and inflammatory cell infiltration, and follicular atresia in ovarian tissue in the 5^th week after operation. From the 5^th to 10^th weeks, the inflammation-associated cytokines TGF-β and IL-4 were decreased, while IFN-γ and IL-21 were elevated in D3tx mice. In the spleen and local nodes of the D3 tx group, CD4^+CD25^+CD127^low/- Treg cells increased slightly in 1-3 weeks, then gradually decreased, and were lower than that of the control group 5 weeks later;CD138-IgD+cells were decreased in 1-10 weeks, while the CD138^+ CD19^-IgD^- cells were increased in 5-10 weeks(P<0.05). In conclusion, the temporal changes of immune function in AOD patients are closely related to the occurrence and development of AOD;and there is a clear window period from the occurrence of immune dysfunction to ovarian histological changes(1-4 weeks after surgery), which suggests that intervention in the window phase and improvement of immune function may improve the outcome of autoimmune ovarian disease.