中华儿科杂志
中華兒科雜誌
중화인과잡지
Chinese Journal of Pediatrics
2009年
11期
856-861
,共6页
尹晓娟%刘冬云%罗分平%龙琦%封志纯
尹曉娟%劉鼕雲%囉分平%龍琦%封誌純
윤효연%류동운%라분평%룡기%봉지순
大鼠%Sprague-Dawley%缺氧缺血%脑%成纤维细胞生长因子2%蛋白质生物合成
大鼠%Sprague-Dawley%缺氧缺血%腦%成纖維細胞生長因子2%蛋白質生物閤成
대서%Sprague-Dawley%결양결혈%뇌%성섬유세포생장인자2%단백질생물합성
Rats,Sprague-Dawley%Hypoxic-ischemic brain damage%Fibroblast growth factor 2%Protein biosynthesis
目的 探讨碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)对新生鼠HIBD骨形态发生蛋白4蛋白及其mRNA表达的影响.方法 新生7日SD乳鼠120只,随机分①bFGF组、②HIBD组、③正常对照组,每组40只.HIBD模型建立后①组予以bFGF干预,腹腔注射,连用5 d,根据处死时相点各组又分为7、14、21、28 d等4个小组.采用免疫组化技术检测各组骨形态发生蛋白4(BMP4)蛋白在海马的表达;采用原位杂交技术检测各组BMP4 mRNA在海马的表达;采用TUNEL实验检测各组神经细胞的凋亡.采用随机组设计资料的方差分析进行组间及组内比较.结果 在7 d和14 d时相点,bFGF组BMP4蛋白在海马的表达较H1BD组多;2组在14 d时相点BMP4蛋白在海马的表达较7 d时相点弱.21 d小组、28 d小组等BMP4蛋白在海马的表达3组之间数量无明显变化.在7 d时相点,HIBD组、bFGF组CA1区BMP4 mRNA表达明显,较正常对照组明显增加;在14 d时相点,HIBD组BMP4 mRNA在病侧海马广泛表达,bFGF组仅病侧海马CA1区BMP4 mRNA表达明显,但2组较正常对照组明显增加;21 d时相点BMP4 mRNA在HIBD组、bFGF组海马的表达均显著;28 d时相点BMP4 mRNA在HIBD组病侧海马的表达开始减少,而bFGF组BMP4mRNA在病侧海马的表达无变化.bFGF组凋亡神经细胞在7 d时相点较HIBD组少[(20.10±0.35)vs(29.12±0.31);F=9.010,P<0.01];在14、21、28 d 3个时相点,HIBD组和bFGF组凋亡神经细胞均较7 d时相点增多,bFGF组凋亡神经细胞仍较HIBD组少[(28.09±0.26) vs (37.46±0.23);(18.75±0.71) vs (35.36±0.77);(12.26±0.57) vs (25.70±0.21);F=9.202,7.932,14.985,P<0.01].结论 bFGF对HIBD具有神经修复作用,其作用机制足促进BMP4蛋白及其mRNA在新生鼠HIBD海马的表达,并抑制神经细胞的凋亡.
目的 探討堿性成纖維細胞生長因子(basic fibroblast growth factor,bFGF)對新生鼠HIBD骨形態髮生蛋白4蛋白及其mRNA錶達的影響.方法 新生7日SD乳鼠120隻,隨機分①bFGF組、②HIBD組、③正常對照組,每組40隻.HIBD模型建立後①組予以bFGF榦預,腹腔註射,連用5 d,根據處死時相點各組又分為7、14、21、28 d等4箇小組.採用免疫組化技術檢測各組骨形態髮生蛋白4(BMP4)蛋白在海馬的錶達;採用原位雜交技術檢測各組BMP4 mRNA在海馬的錶達;採用TUNEL實驗檢測各組神經細胞的凋亡.採用隨機組設計資料的方差分析進行組間及組內比較.結果 在7 d和14 d時相點,bFGF組BMP4蛋白在海馬的錶達較H1BD組多;2組在14 d時相點BMP4蛋白在海馬的錶達較7 d時相點弱.21 d小組、28 d小組等BMP4蛋白在海馬的錶達3組之間數量無明顯變化.在7 d時相點,HIBD組、bFGF組CA1區BMP4 mRNA錶達明顯,較正常對照組明顯增加;在14 d時相點,HIBD組BMP4 mRNA在病側海馬廣汎錶達,bFGF組僅病側海馬CA1區BMP4 mRNA錶達明顯,但2組較正常對照組明顯增加;21 d時相點BMP4 mRNA在HIBD組、bFGF組海馬的錶達均顯著;28 d時相點BMP4 mRNA在HIBD組病側海馬的錶達開始減少,而bFGF組BMP4mRNA在病側海馬的錶達無變化.bFGF組凋亡神經細胞在7 d時相點較HIBD組少[(20.10±0.35)vs(29.12±0.31);F=9.010,P<0.01];在14、21、28 d 3箇時相點,HIBD組和bFGF組凋亡神經細胞均較7 d時相點增多,bFGF組凋亡神經細胞仍較HIBD組少[(28.09±0.26) vs (37.46±0.23);(18.75±0.71) vs (35.36±0.77);(12.26±0.57) vs (25.70±0.21);F=9.202,7.932,14.985,P<0.01].結論 bFGF對HIBD具有神經脩複作用,其作用機製足促進BMP4蛋白及其mRNA在新生鼠HIBD海馬的錶達,併抑製神經細胞的凋亡.
목적 탐토감성성섬유세포생장인자(basic fibroblast growth factor,bFGF)대신생서HIBD골형태발생단백4단백급기mRNA표체적영향.방법 신생7일SD유서120지,수궤분①bFGF조、②HIBD조、③정상대조조,매조40지.HIBD모형건립후①조여이bFGF간예,복강주사,련용5 d,근거처사시상점각조우분위7、14、21、28 d등4개소조.채용면역조화기술검측각조골형태발생단백4(BMP4)단백재해마적표체;채용원위잡교기술검측각조BMP4 mRNA재해마적표체;채용TUNEL실험검측각조신경세포적조망.채용수궤조설계자료적방차분석진행조간급조내비교.결과 재7 d화14 d시상점,bFGF조BMP4단백재해마적표체교H1BD조다;2조재14 d시상점BMP4단백재해마적표체교7 d시상점약.21 d소조、28 d소조등BMP4단백재해마적표체3조지간수량무명현변화.재7 d시상점,HIBD조、bFGF조CA1구BMP4 mRNA표체명현,교정상대조조명현증가;재14 d시상점,HIBD조BMP4 mRNA재병측해마엄범표체,bFGF조부병측해마CA1구BMP4 mRNA표체명현,단2조교정상대조조명현증가;21 d시상점BMP4 mRNA재HIBD조、bFGF조해마적표체균현저;28 d시상점BMP4 mRNA재HIBD조병측해마적표체개시감소,이bFGF조BMP4mRNA재병측해마적표체무변화.bFGF조조망신경세포재7 d시상점교HIBD조소[(20.10±0.35)vs(29.12±0.31);F=9.010,P<0.01];재14、21、28 d 3개시상점,HIBD조화bFGF조조망신경세포균교7 d시상점증다,bFGF조조망신경세포잉교HIBD조소[(28.09±0.26) vs (37.46±0.23);(18.75±0.71) vs (35.36±0.77);(12.26±0.57) vs (25.70±0.21);F=9.202,7.932,14.985,P<0.01].결론 bFGF대HIBD구유신경수복작용,기작용궤제족촉진BMP4단백급기mRNA재신생서HIBD해마적표체,병억제신경세포적조망.
Objective To investigate the effect of basic fibroblast growth factor (bFGF) on expression of protein and mRNA of bone morphogenetic protein 4 in hypoxic-ischemic brain damage (HIND) in newborn rats. Method One hundred and twenty 7 days old neonatal rats were randomly divided into control group, hypoxic-ischemic brain damage and interventional group of bFGF, each having forty neonatal rats. After HIBD model was established, bFGF was given to interventional group by peritoneal injection for 5 continuous days. Every group was randomly divided into 7 days, 14 days, 21 days and 28 days group, according to the time of sacrifice. BMP4 protein in hippocampus was determined with immunohistochemical method. Messenger RNA of BMP4 were determined with in situ hybridization. Apoptosis of nerve cell was determined with TUNEL. Intergroup or intragroup comparisons were performed with analysis of variance. Result On the days 7 and 14, expression of BMP4 protein in hippocampus was higher in interventional group of bFGF than in HIBD while expression of BMP4 protein in interventional group of bFGF and HIBD was lower on day 7 than on day 14. Expression of BMP4 protein on the days 21 and 28 had no significant difference among three groups, mRNA expression of BMP4 in interventional group of bFGF and HIBD was significantly higher in hippocampus than in control group. On the day 14, BMP4 mRNA in hippocampus widely expressed in HIBD while BMP4 mRNA only expressed in CA1 in interventional group of bFGF. Expression of BMP4 mRNA in hippocampus on the affected side decreased from the time of killing on 28th day while there was no significant change in interventional group of bFGF. Apoptosis of neural cells at the time of sacrifice on day 7 was lower in interventional group of bFGF than that in HIBD group(F=9.010, P <0.01). Apoptotic neural cells was higher in bFGF and HIBD groups at the time of killing on days 14, 21 and 28 than that on day 7 but that the bFGF group had less apoptotic neural cells than HIBD group(F= 9.202, 7.932, 14.985, P < 0.01). Conclusions bFGF has a neurorestoration effect, which promotes expression of BMP4 protein and BMP4 mRNA in hippocampus of HIBD and inhibit apoptosis of neural cells.
