LPS致奶牛蹄真皮细胞炎症模型的建立
发布时间：2018-07-23 09:09
【摘要】：为建立LPS致奶牛蹄真皮细胞炎症模型,通过分离、培养奶牛蹄真皮细胞,用不同浓度的LPS处理蹄真皮细胞,倒置显微镜下观察细胞形态,MTT法检测细胞活力,RT-PCR法检测CYP3A4、CYP1A1mRNA的表达,Western blot法检测CYP450蛋白的表达,并通过ELISA法检测细胞上清液中TNF-ɑ、IL-1β的含量,来确定LPS是否对细胞造成损伤。结果表明,随着LPS浓度的升高,对蹄真皮细胞的损伤也随之升高,CYP450蛋白的表达呈升高的趋势,在10μg/mL时表达最明显,10μg/mL LPS处理48h,对CYP3A4、CYP1A1mRNA表达的抑制作用较明显,模型组TNF-ɑ和IL-1β的含量极显著高于对照组(P0.01)。本试验表明,10μg/mL LPS作用48h可建立可靠的蹄真皮细胞炎症模型。
[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.
【作者单位】： 河北农业大学动物医学院;
【基金】：河北省硕士研究生创新资助项目“水飞蓟对LPS诱导的奶牛蹄真皮炎症细胞的影响”(CXZZSS2017067) 河北省现代农业产业技术体系奶牛产业创新团队建设项目“奶牛疾病防控技术研发与示范”(HBCT2013080204)
【分类号】：S858.23
本文编号：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.
【作者单位】： 河北农业大学动物医学院;
【基金】：河北省硕士研究生创新资助项目“水飞蓟对LPS诱导的奶牛蹄真皮炎症细胞的影响”(CXZZSS2017067) 河北省现代农业产业技术体系奶牛产业创新团队建设项目“奶牛疾病防控技术研发与示范”(HBCT2013080204)
【分类号】：S858.23
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