大叶枸浸出液对WIRS小鼠胃及睾丸的保护作用及NOS的表达测定

发布时间：2018-04-21 19:25

本文选题：大叶枸浸出液 + 应激　；参考：《南京农业大学》2015年硕士论文

【摘要】：在应激情况下,动物会产生采食量降低,生产性能改变以及繁殖效率的降低。研究表明饮食中添加维生素、氨基酸、微量物质及中药等,对减轻机体的应激伤害具有重要的意义。因此,寻求更有效、安全及经济的药物具有重要意义。大叶枸是应用细胞工程技术培育出的新型植株,为桑科多年生落叶乔木,在农业生产中具有很高的营养价值。桑科植物具有丰富的植物素可以治疗许多疾病,例如,降低由氧化应激引起的睾酮耗竭,改善由肥胖引起的炎症,抗高血糖以及通过降低NO(由iNOS介导产生)的水平来诱导癌细胞凋亡。水浸束缚型应激(Water immersion and restraint stress,WIRS)已经作为一个可以诱导可复制的应激模型被许多的研究人员在不同组织中运用。但是,对于大叶枸在WIRS介导的胃及睾丸损伤中的作用还没有报道。一氧化氮(NO)是由一氧化氮合酶(NOS)合成的具有高活性的亲脂质小分子,几乎产生于机体所有的组织和器官,参与许多生理学及病理学的生物进程,并介导着细胞毒及细胞间的信号传递或称信使作用。本文以WIRS小鼠为模型,研究了大叶枸浸出液对WIRS诱导的胃及睾丸的影响及NOS的生理作用,并对无应激组、应激3 h组及应激恢复组的小鼠胃及睾丸总抗氧化能力(T-AOC)、NOS活性及睾丸组织的基因表达水平进行了检测,另外,本试验采用组织学及免疫组织化学技术对WIRS小鼠胃及睾丸组织损伤及NOS的蛋白定位进行了检测,得到以下结果。1.WIRS对胃及睾丸的损伤结果表明,经过3 h的应激作用,可以对胃及睾丸组织造成一定的损伤,而经过10d的恢复作用,损伤基本痊愈。另外,添加大叶枸浸出液的小鼠胃及睾丸组织的损伤程度明显轻于对照组。2.WIRS小鼠胃及睾丸组织T-AOC及NOS活性水平WIRS可以提高胃组织iNOS水平,降低T-AOC及cNOS水平;而提前饲喂大叶枸浸出液可以降低胃组织iNOS水平,提高cNOS及T-AOC水平。对于睾丸组织,WIRS可以降低T-AOC的活性水平,而提前饲喂大叶枸浸出液可以提高T-AOC的水平。但是NOS活性水平在睾丸应激前后及是否饲喂大叶枸浸出液没有显著变化。3.NOS在胃及睾丸组织上的蛋白定位胃组织免疫组织化学结果表明,iNOS及eNOS浓染于胃壁细胞,而相反,nNOS在胃壁细胞上的染色比较弱。除此之外,只有iNOS及eNOS在胃组织血管中有表达,nNOS未见表达。睾丸组织免疫组织化学结果表明,iNOS及eNOS除了在间质细胞及支持细胞表达外还表达在精细胞、精子及血管中;而nNOS在此三种组织中未见表达。但是,nNOS却强表达于间质细胞,提示nNOS在类固醇激素的产生中扮演着重要角色。4.NOS mRNA在睾丸中的表达结果显示WIRS小鼠睾丸iNOS及eNOS基因表达量升高,然而饲喂大叶枸浸出液的小鼠显示降低的iNOS、eNOS基因水平。综上述,WIRS可以对胃及睾丸组织造成一定的损伤,而大叶枸浸出液对WIRS诱导的胃及睾丸组织损伤具有保护作用,iNOS及eNOS在WIRS小鼠胃及睾丸组织生理机制中发挥重要作用。然而,NOS各亚型在其中的具体调控机制还需要进一步研究。
[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.

【学位授予单位】：南京农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S816.4

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