亚洲免费av电影一区二区三区,日韩爱爱视频,51精品视频一区二区三区,91视频爱爱,日韩欧美在线播放视频,中文字幕少妇AV,亚洲电影中文字幕,久久久久亚洲av成人网址,久久综合视频网站,国产在线不卡免费播放

        ?

        Pten基因?qū)δ膛H橄偕掀ぜ毎谌榈恼{(diào)節(jié)功能

        2016-10-21 04:23:01王卓然王春梅王杰李慶章高學軍
        科技創(chuàng)新導報 2016年8期
        關鍵詞:泌乳發(fā)育

        王卓然 王春梅 王杰 李慶章 高學軍

        摘 要:該研究以泌乳中期的奶牛乳腺上皮細胞為模型,探索Pten基因的表達與奶牛乳腺發(fā)育和泌乳之間的關系,目的在于揭示Pten基因在奶牛乳腺中的調(diào)節(jié)作用,為動物乳腺發(fā)育和泌乳調(diào)節(jié)機制的研究提供基礎資料。該研究以中國荷斯坦奶牛作為實驗動物,應用熒光定量qRT-PCR、Westernblotting和免疫組織化學技術,對奶牛不同發(fā)育階段及不同乳品質(zhì)的乳腺組織中Pten mRNA和蛋白質(zhì)的相對表達量進行檢測;以體外培養(yǎng)的泌乳中期奶牛乳腺上皮細胞為研究對象,構建重組質(zhì)粒pGCMV-Pten-IRES-EGFP,對細胞進行瞬時轉(zhuǎn)染,進行Pten基因過表達實驗;應用RNA干擾的方法用Pten siRNA瞬時轉(zhuǎn)染細胞,進行Pten基因抑制實驗。分別用相關試劑盒檢測Pten基因過表達和抑制之后細胞?-酪蛋白、甘油三酯以及乳糖分泌的情況,為了檢測Pten基因?qū)δ膛H橄偕掀ぜ毎盍驮鲋衬芰Φ挠绊?,分別應用CASY-TT細胞分析儀和流式細胞儀檢測細胞活性和細胞周期,采用熒光定量qRT-PCR和Western blotting技術,在mRNA和蛋白水平檢測與泌乳相關的信號通路基因的表達變化;同時,添加外源性催乳素和葡萄糖培養(yǎng)細胞,檢測培養(yǎng)液上清中?-酪蛋白、甘油三酯和乳糖的濃度,以及Pten基因的表達量變化,從而探索Pten基因在催乳素誘導的葡萄糖轉(zhuǎn)化生成乳糖過程中的作用。研究結(jié)果表明,泌乳期Pten基因表達量顯著低于干乳期。與泌乳期低乳品質(zhì)奶牛乳腺相比,泌乳期高乳品質(zhì)的奶牛乳腺組織中Pten mRNA和蛋白表達水平分別降低了30%和40%;Pten基因過表達可抑制奶牛乳腺上皮細胞的活力、增殖能力以及?-酪蛋白、甘油三酯和乳糖的分泌量(P<0.05),使奶牛乳腺上皮細胞中MAPK、CyclinD1、AKT、mTOR、S6K1、STAT5、SREBP1、PPAR?、PRLR、GLUT1的表達下調(diào)(P<0.05),并上調(diào)4EBP1的表達水平(P<0.05);Pten基因抑制實驗表現(xiàn)出相反的結(jié)果;而Pten基因的過表達和抑制對ELF5的表達均無顯著影響(P>0.05)。催乳素的添加能促進細胞分泌?-酪蛋白、甘油三酯和乳糖(P<0.05),并下調(diào)Pten基因表達量(P<0.05);葡萄糖的添加顯著增加了?-酪蛋白和乳糖分泌量(P<0.05),但甘油三酯含量無顯著變化(P>0.05),同時Pten基因的表達量無明顯改變(P>0.05)。綜上所述,Pten基因參與調(diào)節(jié)奶牛乳腺上皮細胞泌乳的過程,負向調(diào)節(jié)細胞的活力、增殖能力和細胞周期,并能抑制奶牛乳腺上皮細胞分泌?-酪蛋白、甘油三酯和乳糖;這種調(diào)節(jié)作用是通過Pten基因靶向調(diào)節(jié)PI3K-AKT信號通路,進而調(diào)節(jié)其他泌乳相關信號通路基因的表達而實現(xiàn)的;同時發(fā)現(xiàn)Pten基因的表達受催乳素的負調(diào)節(jié),但葡萄糖對Pten基因的表達水平無顯著影響。

        關鍵詞:奶牛乳腺 發(fā)育 泌乳 Pten

        Abstract:In the aim of detectting the role of Pten gene in the mammary gland of dairy cow, dairy cows mammary epithelial cells (DCMECs) in mid-lactation period were used as models to investigate the relationship of Pten expression and mammary glands development and lactation, which provides basic data for the study of ruminant mammary gland development and lactation mechanisms, and the theoretical support for milk production and milk quality of the artificial regulation at the same time. In this research, Holstein dairy cows were used as experimental animals, applying to qRT-PCR, Western blotting, and immunofluorescence triple staining technology, Pten mRNA and protein expression at different development stages and various milk qualities of dairy cows mammary gland tissue were detected. Furthermore, DCMECs as research objects in vitro were used to study the function of Pten gene. Recombinant plasmid pGCMV-Pten-IRES-EGFP was constructed and transient transfected into cells to prosue the Pten gene overexpression experiment. Meanwhile, RNAi method was used to transfect Pten siRNA in the Pten gene inhibition experiment. We determined concentrations of β-casein, triglyceride, and lactose following Pten gene overexpression and inhibition by specific kits. To determine whether Pten gene affected DCMEC viability and proliferation, cells were analyzed by CASY-TT and flow cytometry. Genes involved in lactation-related signaling pathways were detected by qRT-PCR and Western blotting. After prolactin and glucose were added to the cell cultures, concentrations of β-casein, triglyceride, and lactose were detected, and Pten gene expression was also assessed. Thus investigating the role of Pten gene in the process of glucose transform into lactose induced by prolactin. In summary, we showed that Pten gene is specifically involved in lactation of dairy cow mammary epithelial cells, and down-regulates DCMEC viability, proliferation ability, and the cell cycle along with β-casein, triglyceride, and lactose secretion. Pten gene targets and regulates the PI3K/AKT pathway, which in turn regulates other lactation-related signaling genes. Moreover, the expression of Pten gene can be down-regulated by prolactin, but the introduction of glucose to culture medium revealed no significant difference in Pten gene expression level in DCMECs.

        Key Words:Dairy cow mammary gland;Development;Lactation;Pten

        閱讀全文鏈接(需實名注冊):http://www.nstrs.cn/xiangxiBG.aspx?id=87257&flag=1

        猜你喜歡
        泌乳發(fā)育
        母豬泌乳量不足的危害及提高措施
        青春期乳房發(fā)育困惑咨詢——做決定
        不來月經(jīng)加上泌乳,說不定是腦子長瘤了
        不來月經(jīng)加上泌乳,說不定是腦子長瘤了
        孩子發(fā)育遲緩怎么辦
        中華家教(2018年7期)2018-08-01 06:32:38
        不同泌乳階段駝乳理化指標和體細胞數(shù)的測定分析
        刺是植物發(fā)育不完全的芽
        冰川是發(fā)育而來的
        奧秘(2015年4期)2015-09-10 07:22:44
        西農(nóng)薩能奶山羊泌乳規(guī)律研究*
        中醫(yī)對青春發(fā)育異常的認識及展望
        人妻秘书被社长浓厚接吻| 成l人在线观看线路1| 日韩欧美人妻一区二区三区 | 蜜臀av毛片一区二区三区| 久久精品国产成人午夜福利| 女人高潮内射99精品| 护士人妻hd中文字幕| 香蕉视频www.5.在线观看| 亚洲国产成人无码影院| 国内精品九九久久精品小草| 蜜桃激情视频一区二区| 久久久精品亚洲人与狗| 国产三级精品视频2021| 国产精品永久久久久久久久久| 最爽无遮挡行房视频| 亚洲aⅴ无码成人网站国产app| 国产免费资源| 一区二区三区国产亚洲网站| 成人三级在线| 日本女同伦理片在线观看| 蜜桃视频一区视频二区| 最新国产不卡在线视频| 亚洲开心婷婷中文字幕| 国语对白嫖老妇胖老太| 久久精品夜夜夜夜夜久久| 婷婷激情六月| 九九久久精品一区二区三区av | 蜜桃臀av一区二区三区| 美女mm131爽爽爽| 国产在线无码一区二区三区| 国产精品亚洲片夜色在线| 亚洲国产精品一区亚洲国产| 国产黄久色一区2区三区| 久久精品国产99国产精品亚洲| 人妻少妇边接电话边娇喘| 无码中文字幕在线DVD| 亚洲精品国产不卡在线观看| 亚洲av推荐网站在线观看| 国产一级内射视频在线观看| 女人张开腿让男桶喷水高潮| 欧美高大丰满freesex|