中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2011年
5期
364-369
,共6页
谢涵%邹丽%朱剑文%杨昀%钟少平%王乾华
謝涵%鄒麗%硃劍文%楊昀%鐘少平%王乾華
사함%추려%주검문%양윤%종소평%왕건화
神经生长因子类%肿瘤抑制蛋白质类%基因沉默%新生血管化,生理性%胎盘
神經生長因子類%腫瘤抑製蛋白質類%基因沉默%新生血管化,生理性%胎盤
신경생장인자류%종류억제단백질류%기인침묵%신생혈관화,생이성%태반
Nerve growth factors%Tumor suppressor proteins%Gene silencing%Neovascularization,physiologic%Placenta
目的 研究轴突导向因子netrin-1基因沉默对胎盘血管生成的影响.方法 应用RNA 干扰技术分别沉默netrin-1基因在人脐静脉内皮细胞(HUVEC)和孕鼠胎盘组织中的表达;用四甲基偶氮唑蓝(MTT)比色法检测细胞活性;克隆形成实验检测细胞增殖能力;细胞迁移实验检测细胞迁移能力;小管形成实验检测细胞管腔形成能力;观察胎鼠体质量;CD34因子的免疫组化链霉亲和素-生物素-过氧化物酶复合物(SABC)法检测孕鼠胎盘的微血管密度.结果 (1)HUVEC:沉默netrin-1基因后,HUVEC细胞活性受到抑制,克隆形成率由(69±6)%降至(46±5)%,迁移细胞数由(86±17)个降至(46±13)个,小管形成的分支点数由(37±9)个降至(17±5)个,HUVEC的netrin-1基因沉默前后比较,差异均有统计学意义(P＜0.05).(2)孕鼠胎盘组织:沉默孕鼠胎盘组织netrin-1基因后,胎鼠体质量由(2.39±0.17)g降至(2.12±0.10)g,胎盘微血管密度由(258±38)条/mm2降至(197±32)条/mm2,孕鼠胎盘组织netrin-1基因沉默前后比较,差异均有统计学意义(P＜0.05).结论 沉默netrin-1基因,可以抑制体外HUVEC的细胞活性及增殖、迁移和管腔形成能力,同时使体内胎盘组织的血管生成能力下降.netrin-1是重要的促胎盘血管生长基因.
目的 研究軸突導嚮因子netrin-1基因沉默對胎盤血管生成的影響.方法 應用RNA 榦擾技術分彆沉默netrin-1基因在人臍靜脈內皮細胞(HUVEC)和孕鼠胎盤組織中的錶達;用四甲基偶氮唑藍(MTT)比色法檢測細胞活性;剋隆形成實驗檢測細胞增殖能力;細胞遷移實驗檢測細胞遷移能力;小管形成實驗檢測細胞管腔形成能力;觀察胎鼠體質量;CD34因子的免疫組化鏈黴親和素-生物素-過氧化物酶複閤物(SABC)法檢測孕鼠胎盤的微血管密度.結果 (1)HUVEC:沉默netrin-1基因後,HUVEC細胞活性受到抑製,剋隆形成率由(69±6)%降至(46±5)%,遷移細胞數由(86±17)箇降至(46±13)箇,小管形成的分支點數由(37±9)箇降至(17±5)箇,HUVEC的netrin-1基因沉默前後比較,差異均有統計學意義(P＜0.05).(2)孕鼠胎盤組織:沉默孕鼠胎盤組織netrin-1基因後,胎鼠體質量由(2.39±0.17)g降至(2.12±0.10)g,胎盤微血管密度由(258±38)條/mm2降至(197±32)條/mm2,孕鼠胎盤組織netrin-1基因沉默前後比較,差異均有統計學意義(P＜0.05).結論 沉默netrin-1基因,可以抑製體外HUVEC的細胞活性及增殖、遷移和管腔形成能力,同時使體內胎盤組織的血管生成能力下降.netrin-1是重要的促胎盤血管生長基因.
목적 연구축돌도향인자netrin-1기인침묵대태반혈관생성적영향.방법 응용RNA 간우기술분별침묵netrin-1기인재인제정맥내피세포(HUVEC)화잉서태반조직중적표체;용사갑기우담서람(MTT)비색법검측세포활성;극륭형성실험검측세포증식능력;세포천이실험검측세포천이능력;소관형성실험검측세포관강형성능력;관찰태서체질량;CD34인자적면역조화련매친화소-생물소-과양화물매복합물(SABC)법검측잉서태반적미혈관밀도.결과 (1)HUVEC:침묵netrin-1기인후,HUVEC세포활성수도억제,극륭형성솔유(69±6)%강지(46±5)%,천이세포수유(86±17)개강지(46±13)개,소관형성적분지점수유(37±9)개강지(17±5)개,HUVEC적netrin-1기인침묵전후비교,차이균유통계학의의(P＜0.05).(2)잉서태반조직:침묵잉서태반조직netrin-1기인후,태서체질량유(2.39±0.17)g강지(2.12±0.10)g,태반미혈관밀도유(258±38)조/mm2강지(197±32)조/mm2,잉서태반조직netrin-1기인침묵전후비교,차이균유통계학의의(P＜0.05).결론 침묵netrin-1기인,가이억제체외HUVEC적세포활성급증식、천이화관강형성능력,동시사체내태반조직적혈관생성능력하강.netrin-1시중요적촉태반혈관생장기인.
Objective To study influence on angiogenesis of placenta by gene silencing of netrin-1.Methods Netrin-1 gene in human umbilical vein endothelial cells(HUVEC)and placenta of pregnant rats were silenced by RNA interference.The following methods were used in this study,including the phenytetrazoliumromide(MTT)for viability,clone formation for proliferation,transwell for migration,and tube formation for angiogenesis in vitro.The change of fetal growth was recorded.Placental microvessel density in pregnant rats was measured by immunohistochemical CD34 staining in vivo.Results (1)HUVEC:viability and proliferation of HUVEC were remarkably inhibited by gene silencing of netrin-1.which number of clone formation,migration cell,tube formation were from(69±6)%,86±17,37±9 decreased to(46±5)%,46±13 and 17±5(P＜0.05)respectively.(2)Placenta of pregnant rats:after netrin-1gene silenced,fetal weight were decreased from(2.39 ±0.17)g to(2.12±0.10)g(P＜0.05).Placental microvessel density was decreased from(258±38)/mm2 to(197±32)/mm2 in vivo(P＜0.05).Conclusions Gene silencing of netrin-1 could inhibit viability,proliferation,migration,tubal formation of HUVEC and angiogcnesis of placenta.Netrin-1 plays an important role in regulating angiogenesis in placenta.
