安石榴苷對脂多糖誘導(dǎo)奶牛子宮內(nèi)膜上皮細(xì)胞炎癥損傷的保護(hù)作用(英文)
發(fā)布時(shí)間:2018-05-22 20:09
本文選題:奶牛子宮內(nèi)膜上皮細(xì)胞 + 炎癥因子; 參考:《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2017年06期
【摘要】:目的:評估安石榴苷對脂多糖誘導(dǎo)奶牛子宮內(nèi)膜上皮細(xì)胞炎癥損傷的保護(hù)作用,并初步探討其作用機(jī)制。創(chuàng)新點(diǎn):首次證明安石榴苷對脂多糖誘導(dǎo)奶牛子宮內(nèi)膜上皮細(xì)胞炎癥損傷具有保護(hù)作用,且此作用與核轉(zhuǎn)錄因子κB(NF-κB)和絲裂原活化蛋白激酶(MAPK)信號通路的抑制相關(guān)。方法:用不同濃度的脂多糖(1、10、30、50和100μg/ml)刺激奶牛子宮內(nèi)膜上皮細(xì)胞3、6、9、12和18 h,篩選出建立炎癥損傷的最佳作用濃度和時(shí)間。安石榴苷預(yù)處理細(xì)胞2 h后用脂多糖刺激12 h,用逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(RT-PCR)檢測炎癥因子白細(xì)胞介素1β(IL-1β)、白細(xì)胞介素6(IL-6)、白細(xì)胞介素8(IL-8)及腫瘤壞死因子α(TNF-α)的表達(dá)。用蛋白質(zhì)免疫印跡試驗(yàn)(Western blotting)的方法檢測核因子κB抑制蛋白α(IκBα)、磷酸化的p65、p38、c-Jun氨基末端激酶(JNK)和細(xì)胞外調(diào)節(jié)蛋白激酶(ERK)的表達(dá)水平。結(jié)論:MTT結(jié)果顯示,30μg/ml脂多糖刺激奶牛子宮內(nèi)膜上皮細(xì)胞12 h能夠造成細(xì)胞活力下降和形態(tài)改變(圖2和3)。RT-PCR結(jié)果顯示,安石榴苷預(yù)處理后炎癥因子顯著降低(圖4)。Western blotting結(jié)果顯示,安石榴苷預(yù)處理能顯著抑制IκBα降解以及p65、p38、JNK和ERK的磷酸化表達(dá)水平(圖5和6)。綜上所述,安石榴苷對脂多糖誘導(dǎo)奶牛子宮內(nèi)膜上皮細(xì)胞炎癥損傷具有保護(hù)作用,在治療奶牛子宮內(nèi)膜炎中具有重要價(jià)值。
[Abstract]:Aim: to evaluate the protective effect of amomegranate on inflammatory injury induced by lipopolysaccharide (LPS) in dairy cow endometrial epithelial cells and to explore its mechanism. Innovation: it is the first time to prove the protective effect of amomegranate on lipopolysaccharide induced inflammatory injury of bovine endometrial epithelial cells, which is related to the inhibition of NF- 魏 B NF- 魏 B) and mitogen-activated protein kinase (MAPK) signaling pathway. Methods: bovine endometrial epithelial cells were stimulated with different concentrations of lipopolysaccharide (lipopolysaccharide) at 50 and 100 渭 g / ml for 12 and 18 hours, and the optimal concentration and time for the establishment of inflammatory injury were selected. After pretreatment with amomegranate for 2 h, lipopolysaccharide was used to stimulate cells for 12 h. The expression of inflammatory cytokines IL-1 尾, IL-6, IL-8 and TNF- 偽 were detected by reverse transcription-polymerase chain reaction (RT-PCR). Western blotting assay was used to detect the expression of nuclear factor-kappa B inhibitor, phosphorylated p65, p38, c-Jun amino terminal kinase (JNKK) and extracellular regulated protein kinase (ERK). Conclusion the cell viability and morphological changes were decreased by 30 渭 g/ml lipopolysaccharide stimulation for 12 h (Fig. 2 and 3).RT-PCR results showed that the inflammatory cytokines were significantly decreased after pretreatment with amgranate (4).Western blotting). The pretreatment of amomegranate could significantly inhibit the degradation of I 魏 B 偽 and the phosphorylation of p65, p38, JNK and ERK (Fig. 5 and 6). To sum up, amomegranate has protective effect on lipopolysaccharide induced inflammatory injury of dairy cow endometrial epithelial cells, and has important value in the treatment of dairy cow endometritis.
【作者單位】: Beijing
【基金】:supported by the National Key Technology R&D Program of China(No.2013BAD10B04) the Importation and Development of High-Caliber Talents Project of Beijing Municipal Institutions(No.CIT&TCD20130324),China
【分類號】:S858.23
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本文編號:1923441
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