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目的研究微小隐孢子虫刺激对小鼠腹腔巨噬细胞细胞因子分泌的影响,以及TLR2在微小隐孢子虫刺激小鼠腹腔巨噬细胞中的作用。方法用微小隐孢子虫子孢子刺激野生型(WT)小鼠腹腔巨噬细胞8h,并设置不加任何物质对照组。收取细胞培养上清,采用ELISA法检测IL-6、IL-10、IL-12和TNF-α水平。其次,将微小隐孢子虫子孢子分别与WT小鼠腹腔巨噬细胞和TLR2-/-小鼠腹腔巨噬细胞共培养,收取2、4、8h细胞培养上清,采用ELISA法检测IL-6、IL-10、IL-12、TNF-α水平并进行比较,分析TLR2对微小隐孢子虫刺激小鼠腹腔巨噬细胞分泌细胞因子的影响。结果微小隐孢子虫子孢子刺激WT小鼠腹腔巨噬细胞IL-6(t=10.34)、IL-10(t=55.10)、IL-12(t=10.91)、TNF-α(t=11.078)水平较对照组显著升高(P<0.01);微小隐孢子虫子孢子刺激TLR2-/-小鼠腹腔巨噬细胞与WT小鼠比较IL-6(t8hr=4.299)显著下调(P<0.01),IL-10(t8hr=8.858)和IL-12(t4hr=7.329,t8hr=17.479)显著上调(P<0.01),TNF-α水平差异无统计学意义。结论微小隐孢子虫可以诱导小鼠腹腔巨噬细胞IL-6、IL-10、IL-12、TNF-α分泌水平增高,而TLR2对微小隐孢子虫刺激小鼠腹腔巨噬细胞细胞因子分泌具有调节作用。
Objective To study the effect of Cryptosporidium parvum stimulation on the secretion of cytokines in mouse peritoneal macrophages and the role of TLR2 in the stimulation of mouse peritoneal macrophages by Cryptosporidium parvum. Methods Wild type (WT) mice peritoneal macrophages were stimulated with Cryptosporidium parvum sporozoites for 8 hours, and no substance control group was set. The cell culture supernatants were collected and the levels of IL-6, IL-10, IL-12 and TNF-α were detected by ELISA. Second, the Cryptosporidium parvum sporozoites were co-cultured with peritoneal macrophages and TLR2 - / - mice peritoneal macrophages of WT mice respectively, and the supernatants of 2,4,8h cells were collected. The levels of IL-6, IL-10, IL-12, TNF-α levels were compared and analyzed, TLR2 on Cryptosporidium parvum-stimulated murine peritoneal macrophage secretion of cytokines. Results The levels of IL-6, IL-10 (t = 55.10), IL-12 (t = 10.91) and TNF- α (t = 11.078) in WT mouse peritoneal macrophages were significantly increased (P <0.01). IL-6 (t8hr = 4.299) was significantly lower in TLR2 - / - mice peritoneal macrophages stimulated by C. parvum spores than in WT mice (P <0.01) -10 (t8hr = 8.858) and IL-12 (t4hr = 7.329, t8hr = 17.479) (P <0.01). There was no significant difference in TNF- Conclusions Cryptosporidium parvum can induce the secretion of IL-6, IL-10, IL-12 and TNF-α in mouse peritoneal macrophages to increase, while TLR2 has the inhibitory effect on the secretion of cytokines in peritoneal macrophages stimulated by C. parvum Regulatory effect.