本文選題：大葉枸浸出液 + 應(yīng)激�。� 參考：《南京農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：在應(yīng)激情況下,動(dòng)物會(huì)產(chǎn)生采食量降低,生產(chǎn)性能改變以及繁殖效率的降低。研究表明飲食中添加維生素、氨基酸、微量物質(zhì)及中藥等,對(duì)減輕機(jī)體的應(yīng)激傷害具有重要的意義。因此,尋求更有效、安全及經(jīng)濟(jì)的藥物具有重要意義。大葉枸是應(yīng)用細(xì)胞工程技術(shù)培育出的新型植株,為桑科多年生落葉喬木,在農(nóng)業(yè)生產(chǎn)中具有很高的營(yíng)養(yǎng)價(jià)值。桑科植物具有豐富的植物素可以治療許多疾病,例如,降低由氧化應(yīng)激引起的睪酮耗竭,改善由肥胖引起的炎癥,抗高血糖以及通過(guò)降低NO(由iNOS介導(dǎo)產(chǎn)生)的水平來(lái)誘導(dǎo)癌細(xì)胞凋亡。水浸束縛型應(yīng)激(Water immersion and restraint stress,WIRS)已經(jīng)作為一個(gè)可以誘導(dǎo)可復(fù)制的應(yīng)激模型被許多的研究人員在不同組織中運(yùn)用。但是,對(duì)于大葉枸在WIRS介導(dǎo)的胃及睪丸損傷中的作用還沒(méi)有報(bào)道。一氧化氮(NO)是由一氧化氮合酶(NOS)合成的具有高活性的親脂質(zhì)小分子,幾乎產(chǎn)生于機(jī)體所有的組織和器官,參與許多生理學(xué)及病理學(xué)的生物進(jìn)程,并介導(dǎo)著細(xì)胞毒及細(xì)胞間的信號(hào)傳遞或稱(chēng)信使作用。本文以WIRS小鼠為模型,研究了大葉枸浸出液對(duì)WIRS誘導(dǎo)的胃及睪丸的影響及NOS的生理作用,并對(duì)無(wú)應(yīng)激組、應(yīng)激3 h組及應(yīng)激恢復(fù)組的小鼠胃及睪丸總抗氧化能力(T-AOC)、NOS活性及睪丸組織的基因表達(dá)水平進(jìn)行了檢測(cè),另外,本試驗(yàn)采用組織學(xué)及免疫組織化學(xué)技術(shù)對(duì)WIRS小鼠胃及睪丸組織損傷及NOS的蛋白定位進(jìn)行了檢測(cè),得到以下結(jié)果。1.WIRS對(duì)胃及睪丸的損傷結(jié)果表明,經(jīng)過(guò)3 h的應(yīng)激作用,可以對(duì)胃及睪丸組織造成一定的損傷,而經(jīng)過(guò)10d的恢復(fù)作用,損傷基本痊愈。另外,添加大葉枸浸出液的小鼠胃及睪丸組織的損傷程度明顯輕于對(duì)照組。2.WIRS小鼠胃及睪丸組織T-AOC及NOS活性水平WIRS可以提高胃組織iNOS水平,降低T-AOC及cNOS水平;而提前飼喂大葉枸浸出液可以降低胃組織iNOS水平,提高cNOS及T-AOC水平。對(duì)于睪丸組織,WIRS可以降低T-AOC的活性水平,而提前飼喂大葉枸浸出液可以提高T-AOC的水平。但是NOS活性水平在睪丸應(yīng)激前后及是否飼喂大葉枸浸出液沒(méi)有顯著變化。3.NOS在胃及睪丸組織上的蛋白定位胃組織免疫組織化學(xué)結(jié)果表明,iNOS及eNOS濃染于胃壁細(xì)胞,而相反,nNOS在胃壁細(xì)胞上的染色比較弱。除此之外,只有iNOS及eNOS在胃組織血管中有表達(dá),nNOS未見(jiàn)表達(dá)。睪丸組織免疫組織化學(xué)結(jié)果表明,iNOS及eNOS除了在間質(zhì)細(xì)胞及支持細(xì)胞表達(dá)外還表達(dá)在精細(xì)胞、精子及血管中;而nNOS在此三種組織中未見(jiàn)表達(dá)。但是,nNOS卻強(qiáng)表達(dá)于間質(zhì)細(xì)胞,提示nNOS在類(lèi)固醇激素的產(chǎn)生中扮演著重要角色。4.NOS mRNA在睪丸中的表達(dá)結(jié)果顯示W(wǎng)IRS小鼠睪丸iNOS及eNOS基因表達(dá)量升高,然而飼喂大葉枸浸出液的小鼠顯示降低的iNOS、eNOS基因水平。綜上述,WIRS可以對(duì)胃及睪丸組織造成一定的損傷,而大葉枸浸出液對(duì)WIRS誘導(dǎo)的胃及睪丸組織損傷具有保護(hù)作用,iNOS及eNOS在WIRS小鼠胃及睪丸組織生理機(jī)制中發(fā)揮重要作用。然而,NOS各亞型在其中的具體調(diào)控機(jī)制還需要進(jìn)一步研究。
[Abstract]:Under stress, animals can produce reduced food intake, change of production performance and decrease of reproductive efficiency. The study shows that dietary supplementation of vitamins, amino acids, trace substances and traditional Chinese medicine is of great significance in reducing the stress injury of the body. Therefore, the search for more effective, safe and economic drugs is of great significance. A new type of plant developed by the use of cellular engineering is a high nutritional value of the mulberry perennial deciduous trees in agricultural production. The mulberry plants have rich phytohormones to treat many diseases, such as reducing the exhaustion of testosterone caused by oxidative stress, improving inflammation caused by obesity, anti hyperglycemia, and reducing N by reducing it. O (iNOS mediated) levels induce apoptosis of cancer cells. Water immersion and restraint stress (WIRS) has been used as a replicable stress model for many researchers in different tissues. However, there is no effect on gastric and testicular damage in WIRS mediated gastric and testicular injury. It is reported that nitric oxide (NO) is a highly active small molecule of lipid synthesized by nitric oxide synthase (NOS), which is almost produced in all tissues and organs of the body, participates in many biological processes in physiology and pathology, and mediates cytotoxicity and signal transmission between cells or messenger. This paper studies WIRS mice as a model. The effect of leachate on WIRS induced gastric and testicular formation and the physiological function of NOS, and detection of the total antioxidant capacity (T-AOC), NOS activity and the gene expression level of testicular tissue in the non stress group, stress 3 h group and stress recovery group, and the expression level of NOS activity and testicular tissue were detected. In addition, the test used histology and immunohistochemical technique to W The damage of gastric and testicular tissue in IRS mice and the protein localization of NOS were detected. The results of.1.WIRS damage to the stomach and testis showed that after 3 h stress, the gastric and testicular tissue could be damaged. After the recovery of 10d, the damage of the base was healed. The damage degree of testis tissue was significantly lighter than that of T-AOC and NOS activity level WIRS in the gastric and testicular tissues of the control group.2.WIRS mice, which could improve the iNOS level of the gastric tissue and reduce the level of T-AOC and cNOS, while the early feeding of the leachate of big leaves could reduce the iNOS level of the gastric tissue and improve the cNOS and T-AOC water level. For testicular tissue, WIRS could reduce T-AOC activity. At the level of sex, the level of T-AOC was enhanced by early feeding of the leach solution. But there was no significant change in the activity level of NOS before and after the stress of the testis and in the leach of the leachate. The immunohistochemical results of.3.NOS in the gastric and testicular tissues showed that iNOS and eNOS were concentrated in the gastric wall cells, but on the contrary, nNOS was on the contrary. In addition to this, only iNOS and eNOS were expressed in the blood vessels of the stomach, and the expression of nNOS was not expressed. The immunohistochemical results of the testicular tissue showed that iNOS and eNOS were expressed in spermatocyte, sperm and blood tube except for the expression of interstitial cells and supporting cells, but the expression of nNOS was not expressed in these three tissues. NNOS is strongly expressed in stromal cells, suggesting that nNOS plays an important role in the production of steroid hormones. The expression of.4.NOS mRNA in the testis shows that the expression of iNOS and eNOS in the testis of WIRS mice is elevated. However, mice fed with the leachate from the large leaf extract show a reduced iNOS, eNOS gene level. In a comprehensive way, WIRS can be used for the stomach and testicles. The tissues cause certain damage, and the leachate of big leaf and wolfberry has protective effect on WIRS induced gastric and testicular tissue damage. INOS and eNOS play an important role in the physiological mechanism of gastric and testicular tissue in WIRS mice. However, the specific regulatory mechanism of each subtype of NOS needs to be studied step by step.

【學(xué)位授予單位】：南京農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：S816.4

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