CAJ | 학술논문

目的建立孕早期丙戊酸钠(YPA)暴露致大鼠孤独症谱系障碍(ASD)模型,探究孕早期VPA暴露对仔鼠社会交流行为和神经免疫系统的影响及致病机制.方法 12周龄初孕大鼠分为无暴露组(10只)和VPA暴露组(10只),所生仔鼠分别为正常对照组(10只)和模型组(10只).社会交流测试评价仔鼠是否存在孤独症样社会行为.酶联免疫吸附试验(ELISA)测定孕鼠和仔鼠血清细胞因子白细胞介素(IL)-1β、IL-4、IL-6、干扰素-γ(IFN-γ)和转化生长因子-β(TGF-β)水平,免疫组织化学分析仔鼠脑部小胶质细胞和星形胶质细胞病理改变.结果模型组社会交流测试结果[(-163.16±101.92)分]低于正常对照组[(132.73±114.63)分],差异有统计学意义(t=-6.100,P＜0.05).VPA暴露组母鼠血清细胞因子IL-1β、IL-4、IL-6和IFN-γ水平均高于无暴露组(t=5.883、6.394、5.655、5.393,P均＜0.05),而TGF-β水平低于无暴露组(t=-6.726,P＜0.05).模型组仔鼠血清细胞因子IL-1β、IL-4、IL-6和IFN-y水平均高于正常对照组(t=3.058、3.048、6.670、5.486,P均＜0.05),而TGF-β水平低于正常对照组(t=-6.516,P＜0.05).模型组仔鼠细胞因子水平变化趋势与VPA暴露组母鼠细胞因子水平变化趋势相同.模型组社会交流测试“接近-逃避分数”结果与血清细胞因子IL-1β、IL-4水平呈负相关(r=-0.802、-0.781,P均＜0.05).免疫组织化学结果提示模型组胶质纤维酸性蛋白(GFAP)和钙离子结合调节分子(IBA1)表达高于正常对照组.结论孕早期VPA暴露致孕鼠血清细胞因子水平显著改变,诱发仔鼠免疫激活,免疫功能失衡,脑发育障碍,出现ASD样行为,是孕早期VPA暴露致仔鼠ASD的主要神经免疫学致病机制.
목적 건립잉조기병무산납(YPA)폭로치대서고독증보계장애(ASD)모형,탐구잉조기VPA폭로대자서사회교류행위화신경면역계통적영향급치병궤제.방법 12주령초잉대서분위무폭로조(10지)화VPA폭로조(10지),소생자서분별위정상대조조(10지)화모형조(10지).사회교류측시평개자서시부존재고독증양사회행위.매련면역흡부시험(ELISA)측정잉서화자서혈청세포인자백세포개소(IL)-1β、IL-4、IL-6、간우소-γ(IFN-γ)화전화생장인자-β(TGF-β)수평,면역조직화학분석자서뇌부소효질세포화성형효질세포병리개변.결과 모형조사회교류측시결과[(-163.16±101.92)분]저우정상대조조[(132.73±114.63)분],차이유통계학의의(t=-6.100,P＜0.05).VPA폭로조모서혈청세포인자IL-1β、IL-4、IL-6화IFN-γ수평균고우무폭로조(t=5.883、6.394、5.655、5.393,P균＜0.05),이TGF-β수평저우무폭로조(t=-6.726,P＜0.05).모형조자서혈청세포인자IL-1β、IL-4、IL-6화IFN-y수평균고우정상대조조(t=3.058、3.048、6.670、5.486,P균＜0.05),이TGF-β수평저우정상대조조(t=-6.516,P＜0.05).모형조자서세포인자수평변화추세여VPA폭로조모서세포인자수평변화추세상동.모형조사회교류측시“접근-도피분수”결과여혈청세포인자IL-1β、IL-4수평정부상관(r=-0.802、-0.781,P균＜0.05).면역조직화학결과제시모형조효질섬유산성단백(GFAP)화개리자결합조절분자(IBA1)표체고우정상대조조.결론 잉조기VPA폭로치잉서혈청세포인자수평현저개변,유발자서면역격활,면역공능실형,뇌발육장애,출현ASD양행위,시잉조기VPA폭로치자서ASD적주요신경면역학치병궤제.
Objective To establish the Valproate (VPA) exposure-induced autism spectrum disorder (ASD) model at early pregnancy in rats,and to study the changes of social interaction test and neuroimmune system in autism model rats,and discuss the pathogenic mechanism.Methods Twelve-week-old rats were randomly divided into the non-exposed group (n =10) and the VPA-exposed group (n =10).Their babies were respectively just the normal control group (n =10) and the model group (n =10).The autism-like social behavior was evaluated via the social interaction test.The level of interleukin (IL)-1β,IL-4,IL-6,interferon-gamma (IFN-γ),transforming growth factor-beta (TGF-β) in mothers and offspring were detected by using enzyme-linked immunosorbent assay (ELISA).The histopathological damage in brain was observed with immunohistochemical staining.Results The score of social interaction test in the model group [(-163.16 ± 101.92) scores]was lower than that in the normal control group [(132.73 ± 114.63) scores] (t =-6.100,P ＜ 0.05).The levels of IL-1β,IL-4,IL-6,IFN-γ in VPA-exposed group were higher than those in the non-exposed group (t =5.883,6.394,5.655,5.393,all P ＜ 0.05),while the level of TGF-β was lower than that in the non-exposed group (t =-6.726,P ＜ 0.05).The levels of IL-1β,IL-4,IL-6,IFN-γ in the model group were higher than those in the normal control group(t =3.058,3.048,6.670,5.486,all P ＜ 0.05),while the level of TGF-β was lower than that in the normal control group (t =-6.516,P ＜ 0.05).The change trend of the level of cytokines in the serum of ASD model rats was similar to the change trend of the level of cytokines in the serum of VPA exposed rats.The result of social interaction test "the approach-avoidance score" was inversely correlated with the levels of IL-1β and IL-4 (r =-0.802,-0.781,all P ＜ 0.05).The result of immunohistochemical staining showed the expressions of glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1 (IBA1) in the model group were higher than those in the normal control group.Conclusions The main neuroimmunological pathogenesis mechanism can be explained as follows:at early pregnancy,the maternal cytokines skewing influenced by VPA exposure can make immune activation,induce immune system dysfunction and affect the brain growth and development,which result in autism-like behavior in offspring.