论文部分内容阅读
目的观察播散性毛孢子菌病小鼠模型中白细胞介素-10(IL-10)、转化生长因子β1(TGF-β1)的动态表达,并探讨其在播散性毛孢子菌病中可能发挥的作用。方法 50只雄性BALB/c小鼠随机分为试验组和对照组,每组25只,尾静脉分别接种阿萨希毛孢子菌(T.asahii)悬液和生理盐水,接种后3 d、7 d、14 d、21 d及28 d,对小鼠肾脏载菌量进行测定;取小鼠内眦静脉血,离心获得血清,通过双抗体夹心酶联免疫吸附试验(ELISA)检测两组小鼠不同感染时间血清IL-10和TGF-β1表达水平;实时荧光定量PCR检测两组小鼠不同感染时间脾脏IL-10 m RNA和TGF-β1 m RNA表达情况;采用Pearson相关分析对试验组小鼠不同感染时间肾脏载菌量与血清IL-10、TGF-β1、脾脏IL-10 m RNA、TGF-β1m RNA表达量及试验组小鼠血清IL-10表达水平与脾脏IL-10 m RNA表达量、血清TGF-β1表达水平与脾脏TGF-β1 m RNA表达量进行相关分析。结果接种后3 d,试验组小鼠肾脏载菌量为(69.5±9.1)cfu/mg,接种后7 d,小鼠肾脏载菌量达到峰值,为(120.3±11.4)cfu/mg,其后载菌量逐渐降低,接种后28 d为(2.0±2.5)cfu/mg。试验组小鼠血清IL-10与TGF-β1表达水平在7 d较对照组明显升高,14 d时达到峰值,随后逐渐降低。接种后7d、14 d、21 d,小鼠血清IL-10表达水平与对照组相比,差异有统计学意义(P均<0.05)。接种后7 d、14 d、21 d,小鼠血清TGF-β1表达水平与对照组相比,差异有统计学意义(P均<0.05)。试验组小鼠脾脏IL-10 m RNA、TGF-β1 m RNA表达量在7 d逐渐增高,14 d时达到峰值,随后逐渐降低至对照组水平。接种后7 d、14 d、21 d,小鼠脾脏IL-10 m RNA、TGF-β1 m RNA表达量与对照组相比,差异有统计学意义(P均<0.05)。试验组小鼠血清IL-10表达水平与脾脏IL-10m RNA、血清TGF-β1表达水平与脾脏TGF-β1 m RNA表达量均呈明显正相关(P均<0.05)。接种后7 d、14 d、21 d,试验组小鼠肾脏载菌量与血清IL-10、TGF-β1表达水平及脾脏IL-10 m RNA、TGF-β1 m RNA表达量均呈明显正相关(P均<0.05),而接种后3 d和28 d,试验组小鼠肾脏载菌量与血清IL-10、TGF-β1表达水平及脾脏IL-10 m RNA、TGF-β1 m RNA表达量均无明显相关性(P>0.05)。结论 IL-10和TGF-β1可能参与了小鼠播散性毛孢子菌感染后的免疫应答反应,且表现为先增高后逐渐下降的趋势,可能减弱了机体清除T.asahii的能力,为T.asahii的存活提供了有利条件。
Objective To observe the dynamic expression of interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) in the mouse model of disseminated Coccidio -omycosis and explore the possible role of interleukin-10 Play a role. Methods Fifty male BALB / c mice were randomly divided into experimental group and control group, with 25 rats in each group. The tail vein was inoculated with T.asahii suspension and normal saline respectively. After 3 days and 7 days d, 14 d, 21 d and 28 d, respectively, and then the mice were inoculated into the kidney to determine the amount of bacteriocin-carrying bacteria. The mice were sacrificed and the serum was collected by centrifugation. The two groups of mice were detected by enzyme-linked immunosorbent assay (ELISA) The levels of IL-10 and TGF-β1 mRNA in serum were detected by real-time fluorescence quantitative PCR at different time of infection. The expression of IL-10 mRNA and TGF-β1 mRNA in spleen of mice in different infection groups were detected by real- The expression of IL-10, TGF-β1, spleen IL-10 m RNA and TGF-β1 mRNA in kidney at different times of infection and the level of IL-10 mRNA in spleen , Serum TGF-β1 expression levels and the expression of TGF-β1 m RNA in spleen correlated. Results At 3 days after inoculation, the number of mice in the kidney of mice in the experimental group was (69.5 ± 9.1) cfu / mg and the number of mice in the kidney reached the peak (120.3 ± 11.4) cfu / mg at 7 days after inoculation The amount of inoculated bacteria decreased gradually and reached (2.0 ± 2.5) cfu / mg 28 days after inoculation. The levels of IL-10 and TGF-β1 in the experimental group were significantly higher than those in the control group on the 7th day and reached the peak on the 14th day, then decreased gradually. The levels of IL-10 in mice serum at 7d, 14d and 21d after inoculation were significantly lower than those in control group (all P <0.05). Compared with the control group, the expression of TGF-β1 in the serum of mice on 7 d, 14 d and 21 d after inoculation was significantly different (all P <0.05). The expression of IL-10 mRNA and TGF-β1 m RNA in the spleens of mice in the experimental group increased gradually on the 7th day and peaked on the 14th day, then gradually decreased to the level of the control group. Compared with the control group, the expression of IL-10 mRNA and TGF-β1 m RNA in the spleens of mice on 7d, 14d and 21d after inoculation were significantly different (all P <0.05). The serum level of IL-10 in experimental group was positively correlated with the expression of IL-10m RNA, serum TGF-β1 and TGF-β1 m RNA in spleen (all P <0.05). At 7 d, 14 d, and 21 d after inoculation, the kidney load of mice in test group was positively correlated with the levels of IL-10, TGF-β1 and IL-10 mRNA and TGF-β1 mRNA (P <0.05). At 3 d and 28 d after inoculation, the renal load, IL-10 and TGF-β1 levels in kidney and IL-10 mRNA and TGF-β1 mRNA expression No significant correlation (P> 0.05). Conclusion IL-10 and TGF-β1 may be involved in the immune response to disseminated C. Sp. Infection in mice and may be the first to increase and then decrease. It may reduce the ability of the organism to clear T.asahii, The survival of .asahii provides favorable conditions.