【摘要】：為建立LPS致奶牛蹄真皮細(xì)胞炎癥模型,通過分離、培養(yǎng)奶牛蹄真皮細(xì)胞,用不同濃度的LPS處理蹄真皮細(xì)胞,倒置顯微鏡下觀察細(xì)胞形態(tài),MTT法檢測(cè)細(xì)胞活力,RT-PCR法檢測(cè)CYP3A4、CYP1A1mRNA的表達(dá),Western blot法檢測(cè)CYP450蛋白的表達(dá),并通過ELISA法檢測(cè)細(xì)胞上清液中TNF-ɑ、IL-1β的含量,來確定LPS是否對(duì)細(xì)胞造成損傷。結(jié)果表明,隨著LPS濃度的升高,對(duì)蹄真皮細(xì)胞的損傷也隨之升高,CYP450蛋白的表達(dá)呈升高的趨勢(shì),在10μg/mL時(shí)表達(dá)最明顯,10μg/mL LPS處理48h,對(duì)CYP3A4、CYP1A1mRNA表達(dá)的抑制作用較明顯,模型組TNF-ɑ和IL-1β的含量極顯著高于對(duì)照組(P0.01)。本試驗(yàn)表明,10μg/mL LPS作用48h可建立可靠的蹄真皮細(xì)胞炎癥模型。
[Abstract]:In order to establish an inflammatory model of bovine hoof dermis cells induced by LPS, the hoof dermal cells were cultured and treated with LPS at different concentrations. The expression of CYP450 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of CYP450 protein was detected by Western blot method. The level of TNF-1 尾 in the supernatant was detected by Elisa to determine whether LPS caused damage to the cells. The results showed that with the increase of LPS concentration, the expression of CYP450 protein increased with the increase of LPS concentration. The expression of CYP450 protein was the most obvious at 10 渭 g / mL LPS for 48 h, and the inhibitory effect on CYP1A1 mRNA expression was more obvious at 10 渭 g / mL LPS. The levels of TNF- and IL-1 尾 in the model group were significantly higher than those in the control group (P 0.01). The results showed that 10 渭 g / mL LPS could establish a reliable inflammatory model of hoof dermal cells for 48 h.
【作者單位】： 河北農(nóng)業(yè)大學(xué)動(dòng)物醫(yī)學(xué)院;
【基金】：河北省碩士研究生創(chuàng)新資助項(xiàng)目“水飛薊對(duì)LPS誘導(dǎo)的奶牛蹄真皮炎癥細(xì)胞的影響”(CXZZSS2017067) 河北省現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系奶牛產(chǎn)業(yè)創(chuàng)新團(tuán)隊(duì)建設(shè)項(xiàng)目“奶牛疾病防控技術(shù)研發(fā)與示范”(HBCT2013080204)
【分類號(hào)】：S858.23

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