Wnt2过表达对PINK1 B9 转基因果蝇的作用及机制研究
发布时间：2021-02-16 08:11
目的:探讨Wnt2基因对PINK1B9转基因果蝇（帕金森病模型）的作用及其机制。方法:利用GAL4-UAS系统,构建正常对照组、PINK1B9疾病模型组、Wnt2OE干预组、Wnt2RNAi干预组。第5天时观察各组果蝇的异翅率与飞行率,HE染色观察飞行肌形态,电镜观察线粒体形态,通过Western blot检测Ndufs3、MnSOD、β-catenin、Tubulin蛋白含量,用实时荧光定量PCR法检测线粒体复合物相关基因ND1、ND42、ND75、sdhB、cytb、COX-1,线粒体代谢合成相关基因PGC-1α、NRF1、TFAM及氧化应激相关基因FOXO mRNA表达水平,高分辨率线粒体呼吸测定系统检测ComplexⅠ、ComplexⅡ功能,试剂盒检测ATP、ROS、MDA、GSH相对生成水平。结果:Wnt2过表达降低了PINK1B9转基因果蝇的异翅率（从68.75%±1.18%降低至5.11%±0.65%）,提高了飞行率（从10.79%±1.73%升高至39.35%±2.44%,所有P<0.05）。Wnt...
【文章来源】：桂林医学院广西壮族自治区
【文章页数】：66 页
【学位级别】：硕士
【部分图文】：
PINK1B9转基果蝇中PINK1mRNA表达水平PINK1B9疾病模型组中PINK1mRNA的表达水平极低(ErrorBar表示x±SD，3次独立重复试验
2 A 各组转基因果蝇中 Wnt2 mRNA 的表达情况。UAS-Wnt2OE 与 UAS-Wnt2RNAi 果蝇经HC-GAL4 启动子启动后表现出对应的 Wnt2 mRNA 过表达或降低。( ***表示 P<0.001，各组常对照组比较)。 HA 蛋白 Western blot 图。结果提示 UAS-Wnt2OE 果蝇为正确的 Wnt2 过表达果蝇。分组：a: 对照组，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 各组转基因果蝇中 Wnt2 mRNA 的表达情况。UAS-Wnt2OE 与 UAS-Wnt2RNAi 果HC-GAL4 启动子启动后表现出对应的 Wnt2 mRNA 过表达或降低。( ***表示 P<0.00常对照组比较)。 HA 蛋白 Western blot 图。结果提示 UAS-Wnt2OE 果蝇为正确的 Wnt2 过表达果蝇。分对照组，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.
【参考文献】：
期刊论文
[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)
本文编号：3036477
【文章来源】：桂林医学院广西壮族自治区
【文章页数】：66 页
【学位级别】：硕士
【部分图文】：
PINK1B9转基果蝇中PINK1mRNA表达水平PINK1B9疾病模型组中PINK1mRNA的表达水平极低(ErrorBar表示x±SD，3次独立重复试验
2 A 各组转基因果蝇中 Wnt2 mRNA 的表达情况。UAS-Wnt2OE 与 UAS-Wnt2RNAi 果蝇经HC-GAL4 启动子启动后表现出对应的 Wnt2 mRNA 过表达或降低。( ***表示 P<0.001，各组常对照组比较)。 HA 蛋白 Western blot 图。结果提示 UAS-Wnt2OE 果蝇为正确的 Wnt2 过表达果蝇。分组：a: 对照组，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 各组转基因果蝇中 Wnt2 mRNA 的表达情况。UAS-Wnt2OE 与 UAS-Wnt2RNAi 果HC-GAL4 启动子启动后表现出对应的 Wnt2 mRNA 过表达或降低。( ***表示 P<0.00常对照组比较)。 HA 蛋白 Western blot 图。结果提示 UAS-Wnt2OE 果蝇为正确的 Wnt2 过表达果蝇。分对照组，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.
【参考文献】：
期刊论文
[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)
本文编号：3036477
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