【摘要】：腸道免疫是果蠅抵御外界病原體入侵的重要防線。為研究中藥紫草與三七對果蠅腸道損傷的保護(hù)作用與機(jī)制,利用十二烷基硫酸鈉(sodium dodecyl sulphate, SDS)與百草枯(Paraquat)誘導(dǎo)果蠅腸道損傷,分析果蠅生存率。構(gòu)建由SDS誘導(dǎo)的果蠅腸道損傷模型,通過免疫染色法檢測果蠅腸道前體細(xì)胞數(shù)量、處于分裂期的干細(xì)胞數(shù)量、腸道上皮細(xì)胞內(nèi)活性氧自由基水平與死亡細(xì)胞的數(shù)量,并分析腸道形態(tài)學(xué)變化。實驗結(jié)果表明喂食紫草與三七提取物均可以明顯提高果蠅的生存率、顯著抑制腸道損傷引起的前體細(xì)胞與腸道干細(xì)胞的過度增殖、降低腸道上皮細(xì)胞的死亡；此外,紫草與三七提取物均可以降低腸道上皮細(xì)胞內(nèi)活性氧的水平,保護(hù)和維持腸道形態(tài)。實驗證明,紫草與三七提取物對SDS與Paraquat誘導(dǎo)的果蠅腸道損傷均具有修復(fù)作用。由老齡化引起的干細(xì)胞功能衰退會引發(fā)多種疾病。為了研究果蠅CG12744基因?qū)δc道干細(xì)胞的影響機(jī)制,喂食P araquat、SDS和NaC1檢測CG12744低表達(dá)突變體果蠅生存率,半定量PCR檢測果蠅老齡化過程中各年齡點CG12744蛋白質(zhì)的表達(dá)情況,免疫染色法檢測果蠅老齡化腸道中前體細(xì)胞和處于分裂期的干細(xì)胞的數(shù)量。實驗結(jié)果表明,在喂食Paraquat后有4種CG12744低表達(dá)突變體生存率明顯降低,喂食SDS后有2種CG12744低表達(dá)突變體生存率降低,而在喂食NaC1后CG12744低表達(dá)突變體生存率沒有明顯變化；在果蠅的老齡化過程中CG12744的表達(dá)量呈現(xiàn)增加的趨勢；老齡化過程中,CG12744低表達(dá)突變體中腸內(nèi)出現(xiàn)前體細(xì)胞和干細(xì)胞的過度增殖。實驗結(jié)果證明CG12744基因在果蠅老齡化的過程中起到不可或缺的作用,為進(jìn)一步分析果蠅CG12744基因在老齡化的功能奠定了基礎(chǔ)。
[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.
【學(xué)位授予單位】：東北林業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：Q965

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