视网膜新生血管化/病因学%雌二醇%血管内皮生长因子类%缺氧诱导因子1,α亚基
視網膜新生血管化/病因學%雌二醇%血管內皮生長因子類%缺氧誘導因子1,α亞基
시망막신생혈관화/병인학%자이순%혈관내피생장인자류%결양유도인자1,α아기
Retinal neovascularization/drug therapy%Estradiol%Vascular endothelial growth factors%Hypoxia-inducible factor 1,alpha subunit
目的 观察并探讨17β-雌二醇对高氧诱导的鼠视网膜新生血管形成的影响及机制.方法 48只新生Sprague-Dawley(SD)大鼠随机分为对照组和实验组,每组各24只.对照组大鼠分娩完成后与新生鼠一起正常饲养；实验组大鼠分娩完成后立即与新生鼠一起置于高氧环境饲养.对照组和实验组又再分为磷酸盐缓冲液(PBS)干预组(对照组1、实验1)和雌二醇干预组(对照组2、实验组2),每组各12只大鼠.对照组1和实验组1大鼠分别每日皮下注射PBS0.1 ml;对照组2和实验组2大鼠分别每日皮下注射雌二醇1 μg.每日观察鼠的发育情况.出生后7、14 d逆转录聚合酶链反应(RT-PCR)检测视网膜血管内皮生长因子(VEGF)、低氧诱导因子-1α(HIF-1α)mRNA的含量.出生后14 d时行苏木精-伊红(HE)染色观察视网膜新生血管生长情况；免疫组织化学染色观察VEGF蛋白的表达；透射电子显微镜观察视网膜超微结构变化.结果 出生后14 d各组大鼠标本中突破内界膜的内皮细胞数比较,差异有统计学意义(F=10.7,P＜0.05).其中,实验组1较对照组1明显增多,差异有统计学意义(q=4.28,P＜0.05).实验组2较实验组1明显减少,差异有统计学意义(q=5.16,P＜0.05).实验组2与对照组1比较,差异无统计学意义(q=0.25,P＞0.05).出生后14 d各组大鼠VEGF蛋白表达比较,差异有统计学意义(F=10.7,P＜0.05).其中,实验组1与对照组、实验组2比较,差异有统计学意义(q=5.41,4.35,P＜0.05).视网膜超微结构显示,实验组1神经节细胞肿胀,细胞质淡染,线粒体空泡形成；其他各组视网膜超微结构正常.出生后7、14 d各组大鼠视网膜VEGF、HIF-1 mRNA表达比较,差异均有统计学意义(F=14.7,16.1,13.4,17.5；P=0.001,0.005,0.003,0.009).其中,出生后7d,对照组2 VEGF表达高于对照组1,实验组2VEGF表达高于实验组1,组间比较,差异均有统计学意义(q=5.22,4.32；P＜0.05).出生后14 d,对照组2 VEGF表达高于对照组1,实验组2 VEGF、HIF-1表达低于实验组1,组间比较,差异均有统计学意义(q=3.72,5.12,4.08;P均＜0.05).结论 雌二醇对VEGF mRNA具有双重调控作用,高氧条件下促进视网膜VEGF表达和视网膜血管发育；正常氧条件下通过HIF-1α-VEGF系统抑制视网膜新生血管形成.雌二醇在一定程度上可保护缺氧造成的视网膜超微结构损害.
目的 觀察併探討17β-雌二醇對高氧誘導的鼠視網膜新生血管形成的影響及機製.方法 48隻新生Sprague-Dawley(SD)大鼠隨機分為對照組和實驗組,每組各24隻.對照組大鼠分娩完成後與新生鼠一起正常飼養；實驗組大鼠分娩完成後立即與新生鼠一起置于高氧環境飼養.對照組和實驗組又再分為燐痠鹽緩遲液(PBS)榦預組(對照組1、實驗1)和雌二醇榦預組(對照組2、實驗組2),每組各12隻大鼠.對照組1和實驗組1大鼠分彆每日皮下註射PBS0.1 ml;對照組2和實驗組2大鼠分彆每日皮下註射雌二醇1 μg.每日觀察鼠的髮育情況.齣生後7、14 d逆轉錄聚閤酶鏈反應(RT-PCR)檢測視網膜血管內皮生長因子(VEGF)、低氧誘導因子-1α(HIF-1α)mRNA的含量.齣生後14 d時行囌木精-伊紅(HE)染色觀察視網膜新生血管生長情況；免疫組織化學染色觀察VEGF蛋白的錶達；透射電子顯微鏡觀察視網膜超微結構變化.結果 齣生後14 d各組大鼠標本中突破內界膜的內皮細胞數比較,差異有統計學意義(F=10.7,P＜0.05).其中,實驗組1較對照組1明顯增多,差異有統計學意義(q=4.28,P＜0.05).實驗組2較實驗組1明顯減少,差異有統計學意義(q=5.16,P＜0.05).實驗組2與對照組1比較,差異無統計學意義(q=0.25,P＞0.05).齣生後14 d各組大鼠VEGF蛋白錶達比較,差異有統計學意義(F=10.7,P＜0.05).其中,實驗組1與對照組、實驗組2比較,差異有統計學意義(q=5.41,4.35,P＜0.05).視網膜超微結構顯示,實驗組1神經節細胞腫脹,細胞質淡染,線粒體空泡形成；其他各組視網膜超微結構正常.齣生後7、14 d各組大鼠視網膜VEGF、HIF-1 mRNA錶達比較,差異均有統計學意義(F=14.7,16.1,13.4,17.5；P=0.001,0.005,0.003,0.009).其中,齣生後7d,對照組2 VEGF錶達高于對照組1,實驗組2VEGF錶達高于實驗組1,組間比較,差異均有統計學意義(q=5.22,4.32；P＜0.05).齣生後14 d,對照組2 VEGF錶達高于對照組1,實驗組2 VEGF、HIF-1錶達低于實驗組1,組間比較,差異均有統計學意義(q=3.72,5.12,4.08;P均＜0.05).結論 雌二醇對VEGF mRNA具有雙重調控作用,高氧條件下促進視網膜VEGF錶達和視網膜血管髮育；正常氧條件下通過HIF-1α-VEGF繫統抑製視網膜新生血管形成.雌二醇在一定程度上可保護缺氧造成的視網膜超微結構損害.
목적 관찰병탐토17β-자이순대고양유도적서시망막신생혈관형성적영향급궤제.방법 48지신생Sprague-Dawley(SD)대서수궤분위대조조화실험조,매조각24지.대조조대서분면완성후여신생서일기정상사양；실험조대서분면완성후립즉여신생서일기치우고양배경사양.대조조화실험조우재분위린산염완충액(PBS)간예조(대조조1、실험1)화자이순간예조(대조조2、실험조2),매조각12지대서.대조조1화실험조1대서분별매일피하주사PBS0.1 ml;대조조2화실험조2대서분별매일피하주사자이순1 μg.매일관찰서적발육정황.출생후7、14 d역전록취합매련반응(RT-PCR)검측시망막혈관내피생장인자(VEGF)、저양유도인자-1α(HIF-1α)mRNA적함량.출생후14 d시행소목정-이홍(HE)염색관찰시망막신생혈관생장정황；면역조직화학염색관찰VEGF단백적표체；투사전자현미경관찰시망막초미결구변화.결과 출생후14 d각조대서표본중돌파내계막적내피세포수비교,차이유통계학의의(F=10.7,P＜0.05).기중,실험조1교대조조1명현증다,차이유통계학의의(q=4.28,P＜0.05).실험조2교실험조1명현감소,차이유통계학의의(q=5.16,P＜0.05).실험조2여대조조1비교,차이무통계학의의(q=0.25,P＞0.05).출생후14 d각조대서VEGF단백표체비교,차이유통계학의의(F=10.7,P＜0.05).기중,실험조1여대조조、실험조2비교,차이유통계학의의(q=5.41,4.35,P＜0.05).시망막초미결구현시,실험조1신경절세포종창,세포질담염,선립체공포형성；기타각조시망막초미결구정상.출생후7、14 d각조대서시망막VEGF、HIF-1 mRNA표체비교,차이균유통계학의의(F=14.7,16.1,13.4,17.5；P=0.001,0.005,0.003,0.009).기중,출생후7d,대조조2 VEGF표체고우대조조1,실험조2VEGF표체고우실험조1,조간비교,차이균유통계학의의(q=5.22,4.32；P＜0.05).출생후14 d,대조조2 VEGF표체고우대조조1,실험조2 VEGF、HIF-1표체저우실험조1,조간비교,차이균유통계학의의(q=3.72,5.12,4.08;P균＜0.05).결론 자이순대VEGF mRNA구유쌍중조공작용,고양조건하촉진시망막VEGF표체화시망막혈관발육；정상양조건하통과HIF-1α-VEGF계통억제시망막신생혈관형성.자이순재일정정도상가보호결양조성적시망막초미결구손해.
Objective To investigate the effects and mechanism of 17β-estradiol on the retinal neovasularization in rats with oxygen-induced retinopathy (OIR).Methods A total of 48 SD rats were randomly divided into control group A,control group B,experimental group A and experimental group B with 12 rats in each group.The rats in control group A and experimental group A received a hypodermic injection of 0.1 ml PBS,and the rats in control group B and experimental group B group received an a hypodermic injection of 0.1 ml 17β-estradiol.At postnatal day 7 (P7) and P14,the mRNA expression of vascular endothelial growth factor (VEGF) and Hypoxia-inducible factor (HIF) -1α in the retina were measured by real-time polymerase chain reaction (RT-PCR). At P14,endothelial cell nuclei breaking through the internal limiting membrane were counted after staining with hematoxylin and eosin (HE),and the protein expression of VEGF was measured after immunohistochemical staining.The changes of retinal ultrastructure were observed by transmission electron microscopy.Results At P14,the difference of the number of endothelial cell nuclei among four groups was statistically significant (F=10.7,P＜0.05).The number of endothelial cell nuclei in experimental group A was increased greater than that in control group A (P＜0.05) and experimental group B(q=5.16,P＜0.05).But there was no difference between control group A and experimental group B (q=0.25,P＞0.05).The difference of VEGF protein expression among the four groups was statistically significant (P＜0.05).Comparing experimental group A with control group A,B and experimental group B,the difference was statistically significant (P＜0.05).In experimental group A there was ganglion cell swelling,pale staining cytoplasm and mitochondria vacuolizationin,while these were normal in other three groups.At P7 and P14,the differences of VEGF and HIF-1 mRNA expression among four groups were statistically significant (F=14.7,16.1,13.4,17.5； P=0.001,0.005,0.003,0.009).At P7,the VEGF mRNA expression in control group B was more than that in control group A (q=5.22,P＜0.05).The VEGF mRNA expression in experimental group B was more than that in experimental group A (q=4.32,P＜0.05).At P14,the VEGF mRNA expression in control group B was more than that in control group A (q=3.72,P＜0.05),but there was no difference of HIF-1 mRNA expression between two groups.The VEGF and HIF-1 mRNA expression in experimental group B were both decreased more than those in experimental group A (q =5.12,4.08 ； P＜ 0.05).Conelusions 17β-estradiol has the effect of two-way regulation in VEGF mRNA,which increases VEGF expression in retina under hyperoxic conditions so as to develop the vascular system； which reduces VEGF and HIF-1αexpression so as to prevent pathologic neovascularization under hypoxic conditions. It provides some protection from the damage of retinal neovascularization.
