發(fā)布時(shí)間：2021-02-16 08:11

　　目的:探討Wnt2基因?qū)INK1^B9轉(zhuǎn)基因果蠅（帕金森病模型）的作用及其機(jī)制。方法:利用GAL4-UAS系統(tǒng),構(gòu)建正常對(duì)照組、PINK1^B9疾病模型組、Wnt2OE干預(yù)組、Wnt2RNAi干預(yù)組。第5天時(shí)觀察各組果蠅的異翅率與飛行率,HE染色觀察飛行肌形態(tài),電鏡觀察線粒體形態(tài),通過Western blot檢測Ndufs3、MnSOD、β-catenin、Tubulin蛋白含量,用實(shí)時(shí)熒光定量PCR法檢測線粒體復(fù)合物相關(guān)基因ND1、ND42、ND75、sdhB、cytb、COX-1,線粒體代謝合成相關(guān)基因PGC-1α、NRF1、TFAM及氧化應(yīng)激相關(guān)基因FOXO mRNA表達(dá)水平,高分辨率線粒體呼吸測定系統(tǒng)檢測ComplexⅠ、ComplexⅡ功能,試劑盒檢測ATP、ROS、MDA、GSH相對(duì)生成水平。結(jié)果:Wnt2過表達(dá)降低了PINK1^B9轉(zhuǎn)基因果蠅的異翅率（從68.75%±1.18%降低至5.11%±0.65%）,提高了飛行率（從10.79%±1.73%升高至39.35%±2.44%,所有P<0.05）。Wnt... 

【文章來源】：桂林醫(yī)學(xué)院廣西壯族自治區(qū)

【文章頁數(shù)】：66 頁

【學(xué)位級(jí)別】：碩士

【部分圖文】：

PINK1B9轉(zhuǎn)基果蠅中PINK1mRNA表達(dá)水平PINK1B9疾病模型組中PINK1mRNA的表達(dá)水平極低(ErrorBar表示x±SD，3次獨(dú)立重復(fù)試驗(yàn)

2 A 各組轉(zhuǎn)基因果蠅中 Wnt2 mRNA 的表達(dá)情況。UAS-Wnt2OE 與 UAS-Wnt2RNAi 果蠅經(jīng)HC-GAL4 啟動(dòng)子啟動(dòng)后表現(xiàn)出對(duì)應(yīng)的 Wnt2 mRNA 過表達(dá)或降低。( ***表示 P<0.001，各組常對(duì)照組比較)。 HA 蛋白 Western blot 圖。結(jié)果提示 UAS-Wnt2OE 果蠅為正確的 Wnt2 過表達(dá)果蠅。分組：a: 對(duì)照組，b: UAS-Wnt2OE，c: UAS-Wnt2RNAi.ig.2 (A) Expression level of Wnt2 mRNA. (The graph shows the x ±SD of 3 independentxperiments. Statistical analysis was performed using one-way ANOVA, and the post hoc Bonferroniorrection was also used. *** means P<0.001，each flies compared with the control flies)B)Western blot of HA protein. a: control flies, b: UAS-Wnt2OE，c: UAS-Wnt2RNAi.

2 A 各組轉(zhuǎn)基因果蠅中 Wnt2 mRNA 的表達(dá)情況。UAS-Wnt2OE 與 UAS-Wnt2RNAi 果HC-GAL4 啟動(dòng)子啟動(dòng)后表現(xiàn)出對(duì)應(yīng)的 Wnt2 mRNA 過表達(dá)或降低。( ***表示 P<0.00常對(duì)照組比較)。 HA 蛋白 Western blot 圖。結(jié)果提示 UAS-Wnt2OE 果蠅為正確的 Wnt2 過表達(dá)果蠅。分對(duì)照組，b: UAS-Wnt2OE，c: UAS-Wnt2RNAi.ig.2 (A) Expression level of Wnt2 mRNA. (The graph shows the x ±SD of 3 independentxperiments. Statistical analysis was performed using one-way ANOVA, and the post hoc Bonforrection was also used. *** means P<0.001，each flies compared with the control flies)B)Western blot of HA protein. a: control flies, b: UAS-Wnt2OE，c: UAS-Wnt2RNAi.

【參考文獻(xiàn)】：
期刊論文
[1]Wnt3a:functions and implications in cancer[J]. Sha He,Yi Lu,Xia Liu,Xin Huang,Evan T.Keller,Chao-Nan Qian,Jian Zhang.  Chinese Journal of Cancer. 2015(12)



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