目的：观察大鼠胚胎发育不同时期晶状体的形态变化及缺氧诱导因子-1α( HIF-1α)的表达,探讨HIF-1α在大鼠晶状体胚胎发育过程中的作用。
<br> 方法：清洁级Wistar孕鼠30只,分为6组,分别为胚胎第10,12,14,16,18,20d,每只孕鼠剖腹随机取2只胚鼠,每只胚鼠的一只眼以平行于视神经矢状轴的方向连续切片,常规HE染色,光镜下观察,采用免疫组织化学方法检测HIF-1α的阳性表达；另一只眼采用实时定量PCR方法检测晶状体组织中HIF-1α mRNA的阳性表达。
<br> 结果：大鼠胚胎10d(E10),晶状体泡形成；胚胎12d ( E12),前壁、后壁细胞分化,前壁细胞发育成上皮细胞；胚胎14d(E14),由后壁细胞发育来的原始纤维形成；胚胎16d(E16),上皮细胞增生活跃,次级纤维形成；胚胎20d ( E20),晶状体基本发育成熟。免疫组织化学染色检测到HIF-1α在胚胎期晶状体中高表达,上皮细胞较纤维细胞表达强烈。 E10至E16表达量升高, E16达到峰值,生发区、过渡区染色最强。 E18之后表达量降低,E20最低。6组比较,P<0.01,差异有统计学意义。组与组间两两比较,除E10与E12之间,E14与E16之间,P>0.05,差异无统计学意义外,余各组间比较,P<0.05,差异有统计学意义。实时定量PCR显示,HIF-1α mRNA在胚胎期晶状体中持续表达,且随着胎龄不同变化, E10至 E16, HIF-1αmRNA表达量升高,E16达到峰值,E18、E20下降。6组比较,P<0.01,差异有统计学意义。组与组间两两比较,除E10与E12之间比较, P>0 .05,差异无统计学意义外,余各组间比较,P<0.05,差异有统计学意义。
<br> 结论：在大鼠胚胎10 d时,晶状体泡形成,发育开始,至出生前基本成熟。 HIF-1α在晶状体胚胎发育过程中的阳性表达呈现时空变化。 HIF-1α参与了晶状体的胚胎发育,且在此过程中起重要作用。
目的：觀察大鼠胚胎髮育不同時期晶狀體的形態變化及缺氧誘導因子-1α( HIF-1α)的錶達,探討HIF-1α在大鼠晶狀體胚胎髮育過程中的作用。
<br> 方法：清潔級Wistar孕鼠30隻,分為6組,分彆為胚胎第10,12,14,16,18,20d,每隻孕鼠剖腹隨機取2隻胚鼠,每隻胚鼠的一隻眼以平行于視神經矢狀軸的方嚮連續切片,常規HE染色,光鏡下觀察,採用免疫組織化學方法檢測HIF-1α的暘性錶達；另一隻眼採用實時定量PCR方法檢測晶狀體組織中HIF-1α mRNA的暘性錶達。
<br> 結果：大鼠胚胎10d(E10),晶狀體泡形成；胚胎12d ( E12),前壁、後壁細胞分化,前壁細胞髮育成上皮細胞；胚胎14d(E14),由後壁細胞髮育來的原始纖維形成；胚胎16d(E16),上皮細胞增生活躍,次級纖維形成；胚胎20d ( E20),晶狀體基本髮育成熟。免疫組織化學染色檢測到HIF-1α在胚胎期晶狀體中高錶達,上皮細胞較纖維細胞錶達彊烈。 E10至E16錶達量升高, E16達到峰值,生髮區、過渡區染色最彊。 E18之後錶達量降低,E20最低。6組比較,P<0.01,差異有統計學意義。組與組間兩兩比較,除E10與E12之間,E14與E16之間,P>0.05,差異無統計學意義外,餘各組間比較,P<0.05,差異有統計學意義。實時定量PCR顯示,HIF-1α mRNA在胚胎期晶狀體中持續錶達,且隨著胎齡不同變化, E10至 E16, HIF-1αmRNA錶達量升高,E16達到峰值,E18、E20下降。6組比較,P<0.01,差異有統計學意義。組與組間兩兩比較,除E10與E12之間比較, P>0 .05,差異無統計學意義外,餘各組間比較,P<0.05,差異有統計學意義。
<br> 結論：在大鼠胚胎10 d時,晶狀體泡形成,髮育開始,至齣生前基本成熟。 HIF-1α在晶狀體胚胎髮育過程中的暘性錶達呈現時空變化。 HIF-1α參與瞭晶狀體的胚胎髮育,且在此過程中起重要作用。
목적：관찰대서배태발육불동시기정상체적형태변화급결양유도인자-1α( HIF-1α)적표체,탐토HIF-1α재대서정상체배태발육과정중적작용。
<br> 방법：청길급Wistar잉서30지,분위6조,분별위배태제10,12,14,16,18,20d,매지잉서부복수궤취2지배서,매지배서적일지안이평행우시신경시상축적방향련속절편,상규HE염색,광경하관찰,채용면역조직화학방법검측HIF-1α적양성표체；령일지안채용실시정량PCR방법검측정상체조직중HIF-1α mRNA적양성표체。
<br> 결과：대서배태10d(E10),정상체포형성；배태12d ( E12),전벽、후벽세포분화,전벽세포발육성상피세포；배태14d(E14),유후벽세포발육래적원시섬유형성；배태16d(E16),상피세포증생활약,차급섬유형성；배태20d ( E20),정상체기본발육성숙。면역조직화학염색검측도HIF-1α재배태기정상체중고표체,상피세포교섬유세포표체강렬。 E10지E16표체량승고, E16체도봉치,생발구、과도구염색최강。 E18지후표체량강저,E20최저。6조비교,P<0.01,차이유통계학의의。조여조간량량비교,제E10여E12지간,E14여E16지간,P>0.05,차이무통계학의의외,여각조간비교,P<0.05,차이유통계학의의。실시정량PCR현시,HIF-1α mRNA재배태기정상체중지속표체,차수착태령불동변화, E10지 E16, HIF-1αmRNA표체량승고,E16체도봉치,E18、E20하강。6조비교,P<0.01,차이유통계학의의。조여조간량량비교,제E10여E12지간비교, P>0 .05,차이무통계학의의외,여각조간비교,P<0.05,차이유통계학의의。
<br> 결론：재대서배태10 d시,정상체포형성,발육개시,지출생전기본성숙。 HIF-1α재정상체배태발육과정중적양성표체정현시공변화。 HIF-1α삼여료정상체적배태발육,차재차과정중기중요작용。
AIM: To investigate the morphological changes and the expression of hypoxia-inducible factor-1 alpha ( HIF-1α) subunit during embryonic development of rat lens and explore the role of HIF-1α in lens development process.METHODS: Thirty clean pregnant Wistar rats were divided into 6 embryon groups,10-d, 12-d, 14-d, 16-d, 18-d and 20-d embryo. Two embryons were randomized obtained from every pregnant rat. One of the eyeball samples that were parallel to sagittal axis of optic nerve were cut into serial sections, used HE staining and examined by light microscope. Expression of HIF- 1αprotein in lens was detected by immunohistochemistry. The positive expression of HIF-1α mRNA of the other eyeball samples was detected by real-time PCR.
<br> RESULTS:In the 10 th d of embryo ( E10 ) , the formation of lens vesicle were recognized under the light microscope. In the 12th d of embryo (E12), the anteriorly situated cells and posteriorly situated cells have already differentiated. The anteriorly situated cells were epithelium. In the 14th d of embryo (E14), primary fibers which came from posteriorly situated cells were examined. In the 16th d of embryo ( E16 ), the lens epithelium undergoes extensive proliferation, and enlongate into the secondary fibers. In the 20th d of embryo ( E20 ) , the lens was maturation. By immunohistochemistry staining, the HIF-1α was highly expressed in the lens embryonic development. The expression was gradually promoting from E10 to E16, then reducing. The lens epithelium expressed more HIF-1αthan fibers. The highest mean density was at E16, the lowest at E20. The difference was significant among of the 6 groups (P<0. 0001). The E10 group was combined with the E12 group, the E14 group was combined with the E16 group, showing no significant difference (P>0. 05). The other groups were compared with each other, finding significant difference (P<0. 05). By the real-time PCR, HIF - 1α mRNA was highly expressed in the lens development, and was different at different time. The expression of HIF-1α mRNA was increasing from E10 to E16, then descending at E18 and E20. The expression of HIF-1α mRNA was the highest at E16, the lowest at E20. The difference was significant among of the 6 groups ( P<0. 0001 ). The E10 group was combined with the E12 group, showing no significant difference (P>0. 05). The other groups were compared with each other, finding significant difference (P<0. 05).
<br> CONCLUSION:The lens of Wistar rats differentiate from the E10 when the vesicle formed through the embryo phase. The lens is basic mature before birth. The HIF-1αin the lens embryonic development is highly expressed and changeful. The varies of HIF-1αexpression is depend upon rat embryo development indicating that HIF- 1αmight participate in the process of development of rat lens.
