紫草、三七提取物与CG12744基因对果蝇肠道免疫的影响

发布时间：2018-08-07 14:05

【摘要】：肠道免疫是果蝇抵御外界病原体入侵的重要防线。为研究中药紫草与三七对果蝇肠道损伤的保护作用与机制,利用十二烷基硫酸钠(sodium dodecyl sulphate, SDS)与百草枯(Paraquat)诱导果蝇肠道损伤,分析果蝇生存率。构建由SDS诱导的果蝇肠道损伤模型,通过免疫染色法检测果蝇肠道前体细胞数量、处于分裂期的干细胞数量、肠道上皮细胞内活性氧自由基水平与死亡细胞的数量,并分析肠道形态学变化。实验结果表明喂食紫草与三七提取物均可以明显提高果蝇的生存率、显著抑制肠道损伤引起的前体细胞与肠道干细胞的过度增殖、降低肠道上皮细胞的死亡；此外,紫草与三七提取物均可以降低肠道上皮细胞内活性氧的水平,保护和维持肠道形态。实验证明,紫草与三七提取物对SDS与Paraquat诱导的果蝇肠道损伤均具有修复作用。由老龄化引起的干细胞功能衰退会引发多种疾病。为了研究果蝇CG12744基因对肠道干细胞的影响机制,喂食P araquat、SDS和NaC1检测CG12744低表达突变体果蝇生存率,半定量PCR检测果蝇老龄化过程中各年龄点CG12744蛋白质的表达情况,免疫染色法检测果蝇老龄化肠道中前体细胞和处于分裂期的干细胞的数量。实验结果表明,在喂食Paraquat后有4种CG12744低表达突变体生存率明显降低,喂食SDS后有2种CG12744低表达突变体生存率降低,而在喂食NaC1后CG12744低表达突变体生存率没有明显变化；在果蝇的老龄化过程中CG12744的表达量呈现增加的趋势；老龄化过程中,CG12744低表达突变体中肠内出现前体细胞和干细胞的过度增殖。实验结果证明CG12744基因在果蝇老龄化的过程中起到不可或缺的作用,为进一步分析果蝇CG12744基因在老龄化的功能奠定了基础。
[Abstract]:Intestinal immunity is an important defense line for Drosophila to resist the invasion of external pathogens. In order to study the protective effect and mechanism of traditional Chinese medicine Laver and Panax notoginseng on intestinal injury of Drosophila melanogaster, sodium dodecyl sulfate (sodium dodecyl sulphate, SDS) and paraquat (Paraquat) (Paraquat) were used to induce intestinal injury and survival rate of Drosophila melanogaster was analyzed. The intestinal injury model of Drosophila melanogaster induced by SDS was established. The number of intestinal precursor cells, the number of stem cells in the mitotic phase, the level of reactive oxygen free radicals and the number of dead cells in intestinal epithelial cells were detected by immunostaining. The changes of intestinal morphology were analyzed. The results showed that both the extracts of Rhizoma yezoensis and Panax notoginseng could significantly improve the survival rate of Drosophila melanogaster, significantly inhibit the excessive proliferation of precursor cells and intestinal stem cells induced by intestinal injury, and reduce the death of intestinal epithelial cells. Both the extract of Arnebia chinensis and Panax notoginseng can decrease the level of reactive oxygen species in intestinal epithelial cells and protect and maintain intestinal morphology. The results showed that both the extracts of Rhizoma yezoensis and Panax notoginseng could repair the intestinal injury induced by SDS and Paraquat in Drosophila melanogaster. The decline in stem cell function caused by aging can lead to many diseases. In order to study the effect of CG12744 gene on intestinal stem cells, the survival rate of Drosophila melanogaster with low expression CG12744 was detected by NaC1 and P araquatine SDS, and the expression of CG12744 protein was detected by semi-quantitative PCR at all ages during the aging of Drosophila melanogaster. The number of progenitor cells and mitotic stem cells in the aging intestine of Drosophila melanogaster was detected by immunostaining. The results showed that the survival rates of four CG12744 low expression mutants were significantly decreased after Paraquat feeding, two CG12744 low expression mutants were decreased after SDS feeding, but the survival rates of CG12744 low expression mutants did not change significantly after NaC1 feeding. During the aging of Drosophila melanogaster, the expression of CG12744 increased, and the overproliferation of precursor cells and stem cells appeared in the midgut of low expression mutant CG12744. The results show that CG12744 gene plays an indispensable role in the aging process of Drosophila melanogaster, which lays a foundation for further analysis of the function of Drosophila CG12744 gene in aging.
【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：Q965

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