第58屆Intel ISEF中國(guó)參賽項(xiàng)目：肝細(xì)胞生長(zhǎng)因子對(duì)視網(wǎng)膜色素上皮光損傷的保護(hù)作用
東北育才學(xué)校 高三 李晨
學(xué)科：醫(yī)學(xué)與健康
為探討肝細(xì)胞生長(zhǎng)因子對(duì)視網(wǎng)膜色素上皮光損傷的保護(hù)作用，采用逆轉(zhuǎn)錄PCR從肝組織中得到人肝細(xì)胞生長(zhǎng)因子（hHGF）基因，并將該基因克隆到載體pcDNA3.1（＋）中，構(gòu)建重組真核表達(dá)載體pcDNA－h(huán)HGF，用脂質(zhì)體lipofectamine將載體pcDNA－h(huán)HGF轉(zhuǎn)染體外培養(yǎng)的人視網(wǎng)膜色素上皮（RPE）細(xì)胞，并用ELISA檢測(cè)RPE細(xì)胞培養(yǎng)液中的hHGF，然后將轉(zhuǎn)基因的RPE細(xì)胞和非轉(zhuǎn)基因的RPE細(xì)胞進(jìn)行光照，用單細(xì)胞凝膠電泳方法檢測(cè)RPE細(xì)胞所受到的損傷，比較兩組細(xì)胞損傷的差異。單細(xì)胞凝膠電泳結(jié)果表明，轉(zhuǎn)hHGF基因的視網(wǎng)膜色素上皮細(xì)胞實(shí)驗(yàn)組與非轉(zhuǎn)基因的視網(wǎng)膜色素上皮細(xì)胞對(duì)照組相比，細(xì)胞彗星尾長(zhǎng)及尾/頭明顯變小，其差異具有顯著統(tǒng)計(jì)學(xué)意義(P<0.01)。證實(shí)了肝細(xì)胞生長(zhǎng)因子對(duì)視網(wǎng)膜色素上皮細(xì)胞的光損傷具有一定的保護(hù)作用。
The Protective Effect of HGF against The Light Damage to Retinal Pigment Epithelium
Medcine & Health Sciences
Li Chen 19, Northeast YuCai School
To investigate HGF-mediated protective effect against the light damage to retinal pigment epithelium cells( hRPE ). The cDNA of hHGF was obtained from liver by RT-PCR and inserted into plasmid vecter pcDNA3.1. Then the vecter of pcDNA-hHGF was constructed. The constructed plasmid was transfected into hRPE cells cultured in vitro by a lipfection technology. The expression of hHGF was examined by ELISA technology. Then transfected cultured RPE cells and nontrasfected cultured RPE cells were treated with constant exposure to light.The damage level of both groups was examined by the single cell gel electrophoresis. The result indicats that hHGF transfected hRPE cells significantly survive than nontrasfected hRPE cells. This investigation demostrats that hHGF has significant protective effect against the light damage to retinal pigment epithelium cells.
