米诺环素可以改善新生期BTBR小鼠海马神经发生缺陷  

Minocycline improves hippocampal neurogenetic defects in neonatal BTBR mice

作　　者：骆燚 王刘永葳 郑富杰 宁露 龚洪 范晓棠[1] UO Yi;WANG Liuyongwei;ZHENG Fujie;NING Lu;GONG Hong;FAN Xiaotang(Department of Military Cognitive Psychology,Faculty of Medical Psychology,Army Medical University(Third Military Medical University),Chongqing,400038;Troop 32329 of PLA,Kuqa,Xinjiang Uyghur Autonomous Region,844900;Department of Stomatology,PLA 951 Hospital,Korla,Xinjiang Uyghur Autonomous Region,841000,China)

机构地区：[1]陆军军医大学(第三军医大学)医学心理系军事认知心理学教研室,重庆400038 [2]解放军32329部队,新疆库车844900 [3]解放军第951医院口腔科,新疆库尔勒841000

出　　处：《陆军军医大学学报》2023年第22期2299-2309,共11页Journal of Army Medical University

基　　金：国家自然科学基金青年科学基金项目(82201687)。

摘　　要：目的研究米诺环素(minocycline,Mino)治疗对新生期自闭症模型BTBR T+Itpr3tf/J(BTBR)小鼠海马神经发生的影响。方法将新生健康C57BL/6J(C57)、BTBR小鼠按随机数字表法分为4组:C57生理盐水(C57)组、C57米诺环素(MINO)组、BTBR生理盐水(BTBR)组、BTBR米诺环素(BTBRMINO)组。生后第7天(postnatal day 7,P7),药物处理组小鼠给予米诺环素(22.5 mg/kg)腹腔注射(intraperitoneal injection,i.p.),每天1次,共计8次;C57和BTBR生理盐水组小鼠给予等量的生理盐水。于P14最后1次米诺环素注射2 h后,收取脑组织标本进行免疫荧光染色,检测海马齿状回5-溴脱氧尿嘧啶核苷(5-bromo-2’-deoxyuridine,BrdU)、Ki67、NeuN、DCX、SOX2以及SOX2和GFAP的表达。结果在P14时,相比于C57生理盐水组小鼠,BTBR生理盐水组小鼠海马齿状回BrdU及Ki67标记的神经干细胞增殖数量显著减少(P<0.001),经米诺环素治疗后,BTBR小鼠海马齿状回BrdU(P<0.05)及Ki67(P<0.01)阳性细胞明显增多;BTBR生理盐水组小鼠海马齿状回DCX阳性新生神经元细胞荧光强度显著降低(P<0.001),而NeuN阳性成熟神经元细胞情况类似(P<0.01),米诺环素治疗后,两指标荧光强度显著增强;BTBR生理盐水组小鼠SOX2和GFAP标记的神经干细胞比C57生理盐水组小鼠显著减少(P<0.001),经米诺环素治疗后,该细胞数量显著增多(P<0.01)。两组C57小鼠海马齿状回BrdU、Ki67、NeuN、DCX、SOX2以及SOX2和GFAP的表达差异无统计学意义,表明米诺环素早期干预对C57小鼠海马神经发生没有显著影响。结论米诺环素治疗能显著改善新生期BTBR小鼠海马齿状回神经发生缺陷,促进海马齿状回神经干细胞增殖及神经元分化。Objective To investigate the effects of minocycline(Mino)treatment on hippocampal neurogenesis in early post-life BTBR T+Itpr3tf/J(BTBR)autism model mice.MethodsHealthy newborn C57BL/6J(C57)and BTBR male mice were randomly divided into 4 groups:C57 saline group,C57 Mino group,BTBR saline group,and BTBR Mino group.On the postnatal day 7(P7),the mice of the 2 Mino treatment groups were given intraperitoneal injections of Mino(22.5 mg/kg)once a day for totally 8 times,and those of the other 2 groups were given equal amounts of normal saline.In 2 h after the last injection on P14,the brain samples were collected for immunofluorescence staining to observe the expression of 5-bromo-2’-deoxyuridine(BrdU),Ki67,NeuN,DCX,SOX2,SOX2 and GFAP.Results At P14,the proliferation of BrdU and Ki67 labeled nerve cells(neural stem cells)in the hippocampus dentate gyrus was significantly decreased in the BTBR saline group than the C57 saline group(P<0.001).While,Mino treatment resulted in obviously increased numbers of BrdU(P<0.05)and Ki67(P<0.01)positive cells in the hippocampus dentate gyrus in the BTBR mice.The fluorescence intensity of DCX-positive neonatal neuronal cells was notably decreased in the hippocampal dentate gyrus of BTBR mice(P<0.001),similar result was observed in NeuN-positive cells of mature neurons(P<0.01),and both the intensities could be enhanced markedly by Mino treatment.Though the number of SOX2 and GFAP labeled neural stem cells was significantly lower in the BTBR saline group than the C57 saline group(P<0.001),Mino treatment dramatically increased the number of these neural stem cells(P<0.01).There were no statistical differences in the expression of BrdU,Ki67,NeuN,DCX,SOX2,SOX2 and GFAP in the hippocampal dentate gyrus of the mice between of the C57 saline group and the C57 Mino group,indicating that early Mino intervention had no significant effects on hippocampal neurogenesis of C57 mice.Conclusion Early postnatal Mino treatment significantly improves hippocampal neurogenesis defects in BTBR mice and p

关 键 词：自闭症 新生期小鼠 米诺环素 海马齿状回 神经发生 

分 类 号：R338.26[医药卫生—人体生理学] R749.93[医药卫生—基础医学] R978.14