轮状病毒感染乳鼠后细胞因子水平及肠道外损伤的影响

    Study on cytokines level and extraintestinal injury in suckling mice infected with rotavirus

    • 摘要: 目的:建立BALB/c乳鼠肠道感染轮状病毒(RV)的动物模型,检测血Th1细胞因子Ⅰγ-干扰素(IFN-γ)和Th17细胞因子白细胞介素-17(IL-17)的水平,比较两类细胞因子在不同时间点变化的差异性,进一步探讨RV感染后机体的细胞免疫过程及相关细胞因子的关系,深入了解RV感染的免疫病理特点,为临床防治RV感染及肠道外损伤寻找新的办法。方法:以恒河猴胚肾细胞(MA-104)培养猴RV SA-11株,扩增。实验动物选用清洁级BALB/c乳鼠60只,日龄3~5 d,雌雄不拘,随机分为2组,每组30只,即观察组(RV感染模型组):胃内灌入100 μL(1×10-4)TCD50感染性滴度的SA-11株RV;对照组:胃内灌入100 μL0.9%氯化钠注射液。接种当天以第0天计,每天观察乳鼠情况,收集大便检测RV抗原。分别在接种后的第3天和第8天留取血清检测IFN-γ和IL-17水平。处死乳鼠,留取肝脏、心脏等光镜下观察形态学改变。结果:感染后第3天观察组IFN-γ水平高于对照组(P<0.05),第8天时水平与对照组差异无统计学意义(P>0.05);感染后第3天观察组IL-17水平与对照组比较差异无统计学意义(P>0.05),第8天时水平明显高于对照组(P<0.01)。光镜下观察组乳鼠第3天大部分肝细胞肿胀,中、重度空泡变性,灶状淋巴细胞浸润;心肌细胞浊肿,心肌间质水肿,心内膜、心肌层淋巴细胞浸润,第8天时个别肝脏细胞稍肿胀,轻度空泡变性,心肌细胞大致正常。对照组肝脏及心脏未见明显病理改变。结论:乳鼠感染RV早期以IFN-γ表达为主,且伴有不同程度的肠外脏器损害,但总体上程度轻。而后期IL-17应答占优势,说明IL-17可能在RV的控制上起一定作用,且对肠外脏器的损害有一定修复作用。

       

      Abstract: Objective: To establish the suckling mice model infected with intestinal rotavirus(RV),detect the serum levels of γ-interferon(IFN-γ) and interleukin(IL-17) in suckling mice,compare two kinds of cytokine changes at different time points,further investigate the relationship between cellular immunity and related cytokine,and understand the immunopathological features of RV infection for seeking a new method in preventing RV infection and extraintestinal injury in clinic.Methods: Simian RV strain SA-11 was cultured and amplified from rhesus monkey kidney cells(MA-104).Sixty clean grade BALB/c suckling mice,3 to 5 day-old,half of male and female,were randomly divided into the observation group and control group(30 cases each group).The observation group(RV infection model group) were poured with 100μL(1×10-4) the TCD50 infectious titer of the SA-11 strain RV into stomach,and the control group were poured with 100 μL 0.9% sodium chloride injeetion into stomach.The day of virus inoculation was set as the day 0,and the suckling mice activities were observed after inoculation daily,and the RV antigen was detected in stool.After 3 and 8 days of inoculation,the serum levels of IFN-γ and IL-17 were measured.The suckling mice were sacrificed,and the pathological changes of heart and liver were observed using light microscopy.Results: The serum level of IFN-γ in observation group after 3 days of infection was higher than that in control group(P<0.05),and the difference of which after 8 days of infection between two groups was not statistically significant(P>0.05).The difference of the serum level of IL-17 between two groups after 3 days of infection was not statistically significant(P>0.05),and the serum level of IL-17 in observation group after 8 days of infection was significantly higher than that in control group(P<0.05).Under light microscope,most hepatocytes swelling,median and severe vacuolar degenerating,spotty lymphocytes infiltrating,myocardial cells cloudy swelling,myocardial interstitial edema and endocardial myocardial layer lymphocytes infiltrating were found in observation group after 3 days of infection.After 8 days of infection,the individual liver cells slightly swelling,mild vacuolar degenerating and generally normal myocardial cells were identified in observation group.No obvious changes in liver and heart was found in control group.Conclusions: In intestinal rotavirus infection,especially in early period,the expression of IFN-γ predominates,but it is mild,and the expression of IL-17 predominates in later period.IL-17 may play a role on controlling the RV,and repairing the parenteral organs damage.

       

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