【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(duì)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(cè)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(shí)驗(yàn)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(shí)驗(yàn)證明,紫草與三七提取物對(duì)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(huì)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(cè)CG12744低表達(dá)突變體果蠅生存率,半定量PCR檢測(cè)果蠅老齡化過程中各年齡點(diǎn)CG12744蛋白質(zhì)的表達(dá)情況,免疫染色法檢測(cè)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(shí)驗(yàn)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(shì)；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(shí)驗(yàn)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(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：Q965

【參考文獻(xiàn)】

相關(guān)期刊論文 前9條

1 夏鵬國;張順倉;梁宗鎖;齊志鴻;;三七化學(xué)成分的研究歷程和概況[J];中草藥;2014年17期

2 朱會(huì)超;國晶晶;任煜;;三七總皂苷藥理研究進(jìn)展[J];河南中醫(yī);2014年03期

3 吳穎;孫冰;肖靜;孫桂波;呂圭源;李明;陳素紅;孫曉波;;三七皂苷R1對(duì)LPS誘導(dǎo)的小鼠心肌損傷的保護(hù)作用[J];中國藥理學(xué)通報(bào);2013年02期

4 秦愛萍;張傳印;陸麗;;左旋紫草素抗炎作用的實(shí)驗(yàn)研究[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2009年18期

5 陳祥娜;阮紅;;紫草多糖的免疫調(diào)節(jié)和腫瘤抑制活性研究[J];浙江大學(xué)學(xué)報(bào)(理學(xué)版);2008年06期

6 馬曉東;陳珊珊;李重九;;丙酰紫草素對(duì)煙草花葉病毒外殼蛋白的鈍化作用研究初探[J];分析測(cè)試學(xué)報(bào);2008年S1期

7 周延萌;高允生;;中藥紫草的藥理作用與臨床應(yīng)用[J];醫(yī)藥導(dǎo)報(bào);2008年07期

8 蒲清榮;z丕先;;三七藥理作用研究概述[J];現(xiàn)代醫(yī)藥衛(wèi)生;2007年24期

9 尚西亮;傅華群;劉佳;李學(xué)東;;三七總皂甙對(duì)人肝癌細(xì)胞的抑制作用[J];中國臨床康復(fù);2006年23期

，

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