本文選題：咖啡酸　切入點(diǎn)：脂多糖　出處：《甘肅農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：犬子宮內(nèi)膜炎是寵物臨床上常見的一種產(chǎn)科疾病,嚴(yán)重威脅寵物的健康�？Х人崾且环N具有抗菌抗炎作用的酚酸類化合物。本文通過咖啡酸抗LPS誘導(dǎo)的犬子宮內(nèi)膜上皮細(xì)胞炎癥試驗(yàn),探討咖啡酸的抗炎作用效果,為臨床治療犬子宮內(nèi)膜炎的新藥研發(fā)提供理論依據(jù)。本論文通過酶消法分離培養(yǎng)犬子宮內(nèi)膜上皮細(xì)胞;通過MTT方法檢測LPS對犬子宮內(nèi)膜上皮細(xì)胞細(xì)胞活力的影響,從而篩選出誘導(dǎo)犬子宮內(nèi)膜上皮細(xì)胞炎癥模型的濃度和時間條件;通過MTT方法進(jìn)行咖啡酸的藥物毒性試驗(yàn),確定咖啡酸的安全的劑量范圍和時間,并選擇出高、中、低三個咖啡酸劑量;通過MTT方法測不同劑量咖啡酸對LPS誘導(dǎo)的細(xì)胞活力的影響;通過ELISA方法檢測不同劑量咖啡酸對LPS誘導(dǎo)的細(xì)胞培養(yǎng)液中炎性因子水平的影響;通過一氧化氮試劑盒檢測細(xì)胞培養(yǎng)液中NO濃度的變化;通過AO和EB熒光染料雙染細(xì)胞方法,檢測咖啡酸對LPS誘導(dǎo)的細(xì)胞凋亡的影響。取得試驗(yàn)結(jié)果如下:LPS能夠顯著降低細(xì)胞活力,且呈時間劑量依賴性,50μg/m L LPS作用12 h成功誘導(dǎo)了細(xì)胞炎癥模型;咖啡酸作用12 h后,1 ng/m L~200μg/m L組抑制率與對照組相比沒有顯著差異;咖啡酸作用24 h后,1 ng/m L~25μg/m L組活力無顯著下降。在安全范圍內(nèi)選擇25μg/m L、50μg/m L、100μg/m L作為咖啡酸低、中、高劑量。用中、高咖啡酸預(yù)處理3 h,細(xì)胞活力與對照組相比無顯著差異,比模型組顯著升高。模型組NO、IL-6、TNF-α水平與對照組相比顯著升高;而咖啡酸組則出現(xiàn)不同程度的下降。AO/EB染色后,在熒光顯微鏡下能夠觀察到對照組多數(shù)為活細(xì)胞,模型組有大量凋亡細(xì)胞,而咖啡酸組凋亡細(xì)胞明顯少于模型組。結(jié)果表明,咖啡酸具有一定的抗LPS誘導(dǎo)的犬子宮內(nèi)膜上皮細(xì)胞炎性損傷作用。
[Abstract]:Endometritis is a common obstetrical disease in pets, which is a serious threat to the health of pets.Caffeic acid is a phenolic acid compound with antibacterial and anti-inflammatory effects.In this paper, the anti-inflammatory effect of caffeic acid on canine endometrial epithelial cells induced by LPS was studied, which provides a theoretical basis for the development of new drugs for the treatment of canine endometritis.In this paper, the canine endometrial epithelial cells were isolated and cultured by enzyme digestion, the effects of LPS on the viability of canine endometrial epithelial cells were detected by MTT method, and the concentration and time conditions of the inflammatory model of canine endometrial epithelial cells were screened out.The drug toxicity test of caffeic acid was carried out by MTT method. The safe dose range and time of caffeic acid were determined, and three doses of caffeic acid were selected as high, middle and low doses.The effects of different doses of caffeic acid on the cell viability induced by LPS were measured by MTT method, and the levels of inflammatory factors in cell culture medium induced by LPS were detected by ELISA method.The changes of no concentration in cell culture medium were detected by nitric oxide kit, and the effects of caffeic acid on apoptosis induced by LPS were detected by AO and EB fluorescent dye double staining cell methods.The results were as follows: lipopolysaccharide could significantly reduce cell viability, and the inflammatory model was induced in a dose-dependent manner by 50 渭 g / mL LPS for 12 h, but there was no significant difference in the inhibition rate of 1 ng/m L ~ (200 渭 g / mL) group compared with the control group after 12 h of caffeic acid treatment.There was no significant decrease in activity of 1 ng/m / L 25 渭 g / mL group treated with caffeic acid for 24 h.In the safe range, 25 渭 g / mL 50 渭 g / mL 50 渭 g / mL 100 渭 g / mL caffeic acid was selected as low, medium and high dose of caffeic acid.After treated with high caffeic acid for 3 h, the cell viability was not significantly different from that of the control group, and was significantly higher than that of the model group.Compared with the control group, the level of NO-IL-6 TNF- 偽 in the model group was significantly higher than that in the control group, while in the caffeic acid group, it was observed that most of the cells in the control group were living cells, and a large number of apoptotic cells were found in the model group.The apoptotic cells in caffeic acid group were significantly lower than those in model group.The results showed that caffeic acid could inhibit the inflammatory injury induced by LPS in canine endometrial epithelial cells.
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S858.292

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相關(guān)期刊論文 前1條

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本文編號：1706788