LPS致奶牛蹄真皮細胞炎癥模型的建立
發(fā)布時間：2018-07-23 09:09
【摘要】：為建立LPS致奶牛蹄真皮細胞炎癥模型,通過分離、培養(yǎng)奶牛蹄真皮細胞,用不同濃度的LPS處理蹄真皮細胞,倒置顯微鏡下觀察細胞形態(tài),MTT法檢測細胞活力,RT-PCR法檢測CYP3A4、CYP1A1mRNA的表達,Western blot法檢測CYP450蛋白的表達,并通過ELISA法檢測細胞上清液中TNF-ɑ、IL-1β的含量,來確定LPS是否對細胞造成損傷。結(jié)果表明,隨著LPS濃度的升高,對蹄真皮細胞的損傷也隨之升高,CYP450蛋白的表達呈升高的趨勢,在10μg/mL時表達最明顯,10μg/mL LPS處理48h,對CYP3A4、CYP1A1mRNA表達的抑制作用較明顯,模型組TNF-ɑ和IL-1β的含量極顯著高于對照組(P0.01)。本試驗表明,10μg/mL LPS作用48h可建立可靠的蹄真皮細胞炎癥模型。
[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)大學動物醫(yī)學院;
【基金】：河北省碩士研究生創(chuàng)新資助項目“水飛薊對LPS誘導的奶牛蹄真皮炎癥細胞的影響”(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)新團隊建設(shè)項目“奶牛疾病防控技術(shù)研發(fā)與示范”(HBCT2013080204)
【分類號】：S858.23
本文編號：2138914
[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)大學動物醫(yī)學院;
【基金】：河北省碩士研究生創(chuàng)新資助項目“水飛薊對LPS誘導的奶牛蹄真皮炎癥細胞的影響”(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)新團隊建設(shè)項目“奶牛疾病防控技術(shù)研發(fā)與示范”(HBCT2013080204)
【分類號】：S858.23
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