雌激素膜受体GPR30对手术绝经大鼠全脑缺血后海马神经元凋亡的影响  

EFFECT OF ESTROGEN MEMBRANE RECEPTOR GPR30 ON APOPTOSIS OF HIPPOCAMPAL NEURONS AFTER GLOBAL CEREBAL ISCHEMIA IN POSTMENOPAUSAL RATS

作　　者：白宁 张文丽 李治国 王瑞敏 Bai Ning;Zhang Wenli;Li Zhiguo(Neurobiology Institute of Medical Research Center,North China University of Science and Technology,Tangshan 063210,China)

机构地区：[1]华北理工大学医学实验研究中心神经生物学研究所,河北省唐山市063000

出　　处：《中国煤炭工业医学杂志》2018年第4期420-424,共5页Chinese Journal of Coal Industry Medicine

基　　金：国家自然科学基金(编号:81671223;31171354)

摘　　要：目的研究雌激素膜受体GPR30对手术绝经大鼠全脑缺血后海马CA1区神经元凋亡的影响,并阐明其可能的分子机制。方法 8周龄雌性SD大鼠行双侧卵巢切除以制备手术绝经模型,并随机分为假手术组(Sham)、缺血再灌注组(IR)、GPR30特异激动剂G1处理组(IR+G1)和GPR30特异抑制剂G36处理组(IR+G1+G36);采用四动脉结扎法制备全脑缺血模型(global cerebral ischemia,GCI)、NeuN免疫荧光染色及TUNEL技术观察GPR30对缺血后海马CA1区神经元损伤的影响;Western blot技术检测海马CA1区炎症标志蛋白CD11b的蛋白表达;免疫荧光染色观察GPR30与小胶质细胞标志蛋白Iba1的免疫表达共定位。结果 (1)与Sham组和G1处理组相比,缺血再灌注14d(IR)组和G36处理组海马CA1区NeuN阳性神经元数量显著减少;与之相反,IR组和G36处理组海马CA1区TUNEL阳性染色(凋亡样神经元数量)较Sham组和G1处理组显著增加。(2)IR组和G36处理组可见海马CA1区小胶质细胞标志蛋白Iba1与GPR30的免疫表达共定位。(3)与IR组相比,G1处理可显著降低炎症标志蛋白Iba1、CD11b免疫表达,而G36逆转了以上变化;CD11b的Western blot结果进一步证实G1处理组降低IR14d海马CA1区CD11b表达,G36废弃了G1的作用。结论雌激素膜受体GPR30可降低全脑缺血后海马CA1区神经元损伤,其机制与降低缺血后炎症损伤密切相关。Objective To investigate the effect of estrogen receptor GPR30 on neuronal apoptosis in hipp- ocampal CA1 region after global cerebral ischemia in postmenopausal rats and to elucidate its possible molecular mechanisms. Methods Eight- week- old female SpragueDawley rats were bilaterally ovariectomized to induce the surgical menopause model and were randomly divided into sham operation group, ischemia reperfusion group （ IR）, GPR30 specific agonist G1 treatment group （IR ＋ G1） and GPR30 specific inhibitor G36 treatment group （IR＋ G1 ＋ G36）, nine rats in each group. Global cerebral ischemia （GCI） was induced by four- artery ligation. Immunofluorescence staining was used to detect protein expression of GPR30, NeuN, Ibal or CDllb, and TUNEL technique were used to observe CA1 neuronal apoptosis after ischemia. Western blot analysis was performed to investigate protein expression of inflammatory marker CDllb in hippocampal CA1 region.Results （1） In IR and G36 treatment group animals, NeuN positive staining （number of surviving neurons） was significantly decreased, and TUNEL positive staining （apoptosis- like neurons） contrarily increased compared with sham or G1 - treatment animals in hippocampal CA1 region at 14d reperfusion. （2） In IR and G36 - treated groups, co - localization of GPR30 and microglia marker Ibal was obviously observed in hippocampal CA1 region. （3） Immunofluorescence staining results showed that GI administration significantly attenuated immnuoexpression of Ibal or CDIlb compared to IR group animals, while G36- treatment reversed the change at reperfusion 14d in hippocampal CA1 region. The result of Western blot analysis of CDllb further to confirmed that G1 treatment markedly decreased CDllb protein expression, while G36 a- bolished the effect induced by G1 after 14d reperfusion in hippocampal CA1 region. Conclusion Estrogen receptor GPR30 can reduce the neuronal damage of hippocampal CA1 region after GCI, and its mechanism might be closely related to the red

关 键 词：全脑缺血再灌注 GPR30 神经元 炎症 凋亡 

分 类 号：R364.12[医药卫生—病理学]