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目的建立乳鼠感染的模型,检测轮状病毒(Rotavirus,RV)肠道内、外感染乳鼠肝脏IFN-γ和IL-10水平,比较肝脏IFN-γ和IL-10在轮状病毒肠道内外感染,以及不同时间点变化的差异性,进一步探讨IFN-γ和IL-10免疫自稳失衡与轮状病毒肠道外感染发病的关系,推测以恢复细胞因子平衡为目标的治疗策略可能是治疗轮状病毒肠炎的一种新方法。方法实验动物选用3~5日龄的清洁级BALB/C乳鼠,将54只乳鼠随机分为3组,每组18只,即实验组,包括肠道外组:通过腹腔注入0.10 mL(1×10-5)TCID50感染性滴度计量的SA-11株病毒;肠道内组:通过口腔灌入0.10 mL相同病毒;对照组无特殊处理。感染后,各组动物隔离饲养。观察乳鼠的活动、饮食、体型、毛色和大便变化情况等,收集大便经胶体金法检测其中RV抗原。在接种后的3、5和8 d处死乳鼠,留取肝脏,免疫组化方法检测IFN-γ和IL-10水平。结果对照组2种细胞因子表达量较少,肠道内组IFN-γ水平在接种RV后的第3天明显增多,第3天至第8天缓慢减少;IL-10水平较正常组增高但整个过程未见明显变化。肠道外组IFN-γ水平与肠道内组比较差异无统计学意义;IL-10水平在感染第3天也明显增多,第3天至第8天有所减少但仍存在且高于正常组。结论 BALB/C乳鼠肠道内外感染RV动物模型建立成功。乳鼠肠道外感染早期及后期肝脏内细胞因子呈现出不同改变。所以在肠道外组,肝脏细胞因子平衡机制失衡,进一步阐明这种失衡可能是RV肠道外播散的重要机制。
Objective To establish a model of suckling mice infection and detect the levels of IFN-γ and IL-10 in the liver of swine infected with rotavirus (RV) Infection and different time points to further explore the relationship between the imbalance of immune homeostasis of IFN-γ and IL-10 and the incidence of rotavirus extra-intestinal infections. It is speculated that the therapeutic strategy aiming at restoring the balance of cytokines may be the treatment round A new method for viral enteritis. Methods The experimental animals were selected clean BALB / C suckling mice aged 3 to 5 days. 54 suckling rats were randomly divided into 3 groups with 18 rats in each group, ie experimental group including parenteral group: intraperitoneal injection of 0.10 mL (1 × 10-5) TCID50 infectious titer of SA-11 virus; Intestinal group: 0.10 mL of the same virus was instilled into the oral cavity; no special treatment was given in the control group. After infection, all groups of animals were kept in isolation. Observation of activity, diet, body shape, coat color and stool changes of suckling mice, collecting stool by colloidal gold method to detect RV antigen. The rats were sacrificed at 3, 5, and 8 days after inoculation, the liver was harvested and the levels of IFN-γ and IL-10 were detected by immunohistochemistry. Results The expression levels of two cytokines in the control group were less than those in the control group. The levels of IFN-γ in the gut group were significantly increased on the third day after RV inoculation and slowly decreased on the third to the eighth day. The levels of IL-10 in the gut group were higher than those in the normal group No significant change in the process. The level of IFN-γin the gut group was not significantly different from that in the gut group. The level of IL-10 was also significantly increased on the third day of infection, decreased from the third day to the eighth day but still higher than that in the normal group. Conclusion The animal model of RV infection in BALB / C suckling intestine is established successfully. Early and late stages of parenteral infection in neonatal rat liver cytokines showed different changes. Therefore, in the extragalactic group, the imbalance of hepatic cytokine balance mechanism further clarifies that this imbalance may be an important mechanism of RV pod dispersion.