本文選題：L-茶氨酸 + HO��； 參考：《動(dòng)物營養(yǎng)學(xué)報(bào)》2017年05期

【摘要】：本試驗(yàn)通過建立過氧化氫(H_2O_2)誘導(dǎo)山羊瘤胃上皮傳代細(xì)胞凋亡模型,研究L-茶氨酸對H_2O_2誘導(dǎo)山羊瘤胃上皮傳代細(xì)胞凋亡比率及其凋亡通路關(guān)鍵基因表達(dá)量的影響。選取42日齡湘東黑山羊的瘤胃上皮傳代細(xì)胞,培養(yǎng)于含5%胎牛血清(FBS)的DMEM/F12培養(yǎng)基中,待細(xì)胞密度達(dá)到60%~70%時(shí),隨機(jī)分為5組,分別為培養(yǎng)基中無額外添加物的對照組、添加800μmol/L H_2O_2的Ⅰ組、添加800μmol/L H_2O_2+4 mmol/L L-茶氨酸的Ⅱ組、添加800μmol/L H_2O_2+8 mmol/L L-茶氨酸的Ⅲ組和添加800μmol/L H_2O_2+16 mmol/L L-茶氨酸的Ⅳ組,每組3個(gè)重復(fù)。作用12 h后,應(yīng)用流式細(xì)胞術(shù)(FCM)檢測山羊瘤胃上皮傳代細(xì)胞凋亡比率,并采用實(shí)時(shí)熒光定量PCR(RT-q PCR)對山羊瘤胃上皮傳代細(xì)胞凋亡通路關(guān)鍵基因半胱氨酸天冬氨酸-3(Caspase-3)、半胱氨酸天冬氨酸-8(Caspase-8)、半胱氨酸天冬氨酸-9(Caspase-9)、Fas相關(guān)死亡域蛋白(FADD)和凋亡酶激活因子(Apaf-1)的表達(dá)量進(jìn)行檢測。結(jié)果顯示:1)通過膜聯(lián)蛋白-V(annexin V)/碘化丙啶(PI)聯(lián)合染色結(jié)果可知,與Ⅰ組相比,各L-茶氨酸添加組(Ⅱ、Ⅲ和Ⅳ組)細(xì)胞晚期凋亡比率顯著降低(P0.05),且細(xì)胞晚期凋亡比率隨L-茶氨酸添加濃度的增加逐漸降低。2)通過RT-q PCR檢測結(jié)果可知,與Ⅰ組相比,各L-茶氨酸添加組Caspase-3、Caspase-9、Apaf-1基因的表達(dá)量皆顯著降低(P0.05)。由此得出,L-茶氨酸對H_2O_2引起的山羊瘤胃上皮傳代細(xì)胞凋亡具有一定的保護(hù)作用,該結(jié)果可為今后研究反芻家畜瘤胃氧化應(yīng)激損傷機(jī)制提供技術(shù)支持和理論參考。
[Abstract]:The effect of L-theanine on the rate of apoptosis and the expression of key genes in the apoptosis pathway of rumen epithelial passage of goat induced by H_2O_2 was studied by establishing the model of apoptosis induced by H _ 2O _ 2 (H _ 2O _ 2) in goat rumen epithelium. The rumen epithelial passage cells of 42-day-old Xiangdong black goat were cultured in DMEM/F12 medium containing 5% fetal bovine serum (FBS). When the cell density reached 60 ~ 70, the cells were randomly divided into 5 groups, respectively, as the control group without additional additives in the medium. Group 鈪，

本文編號：1968345