出 处:《北京中医药大学学报》2016年第5期367-371,共5页Journal of Beijing University of Traditional Chinese Medicine
基 金:国家自然科学基金资助项目(No.81573723);教育部博士点博导类基金项目(No.20130013110010)
摘 要:目的比较"截断疗法"和"常规疗法"对流感病毒性肺炎小鼠肺组织炎症反应的作用,以研究"截断疗法"优于"常规疗法"的作用机制,及与病毒感染后机体发生的炎症级联反应间的关系。方法 Balb/c小鼠192只,分为正常组、模型组、常规疗法组和截断疗法组,以50μL 30 LD50流感病毒鼠肺适应株FM1病毒液滴鼻感染后1 h灌胃给药。正常组和模型组以蒸馏水灌胃;常规疗法组第1、2、3天以银翘散水煎剂灌胃,第4、5、6、7天以犀角地黄汤水煎剂灌胃;截断疗法组第1~7天以犀角地黄汤合银翘散水煎剂灌胃;每日给药2次,共给药7 d。以上处理的小鼠分别于第2、4、6、8天摘眼球放血处死并取材、检测。观察肺泡灌洗液(BALF)白细胞总数,酶联免疫吸附测定法(ELISA)检测小鼠肺组织匀浆上清中白细胞介素-1β(IL-1β)和白细胞介素-18(IL-18)水平,实时荧光定量PCR(Real-Time-PCR)检测小鼠肺组织中NLRP3 mRNA的表达。结果截断疗法组和常规疗法组BALF中白细胞总数、IL-1β和IL-18,及NLRP3 mRNA与模型组比较均有不同程度下降,但截断疗法组在第2天BALF中白细胞总数、IL-1β和IL-18水平与模型组比较有显著降低(P<0.01),而常规疗法组尚未显示出明显的作用;第8天截断疗法组白细胞总数明显低于常规疗法组,有显著性差异(P<0.01)。感染后第4天,模型组小鼠肺组织中高表达NLRP3 mRNA,截断疗法组NLRP3 mRNA表达明显减少,常规疗法组与模型组比差异不显著。结论截断疗法在感染早期即可抑制固有免疫应答所诱发的炎症反应,有效阻止了随后的炎症级联反应的发生,截断疾病的传变。其机制可能与抑制NLRP3炎性体的形成、干扰IL-1β和IL-18的成熟和分泌有关。Objective To compare the effects of truncated therapy and conventional therapy on the lung tissue inflammation of mice with pneumonia induced by influenza virus,so as to explore the mechanism of truncated therapy superior to conventional therapy and its relationship with inflammatory cascade after viral infections. Methods 192 Balb / c mice were randomly divided into healthy group,model group,conventional therapy group and truncated therapy group. Except for the healthy group,the mice of the other three groups were infected with 50 μL 30 LD50 mouse lung-adapted influenza virus strain( FM1) by inoculating intranasally. After 1 h of inoculation,healthy group and model group were administered intragastrically( i. g.) distilled water; conventional therapy group was administered i. g. twice daily Yinqiao Powder for the first three days,then Xijiao Dihuang Decoction for the next four days( totae seven days);truncated therapy group was administered i. g. Xijiaodihuang Decoction twice daily for consecutive seven days. Then the mice were sacrificed by taking the eyeballs on the 2nd,4th,6th,and 8thday for sampling and detecting. The WBC count in bronchoalveolar lavage fluid( BALF) was detected,the levels of IL-1βand IL-18 in the supernatants of lung homogenate were measured by ELISA and NOD- like receptor family mem NOD-,LRR-and pyrin domain containing 3( NLRP3) mRNA in the lung tissue were detected by quantitative realtime- PCR. Results Compared with the model group,the WBC counts of BALF,IL-1β,IL-18 and NLRP3 mRNA in truncated therapy group and conventional therapy group decreased( P 0. 01). WBC counts,IL-1β and IL-18 began to show the remarkable differences from that of model group since the 2^(nd) day,while conventional therapy group didn't. On 8thday,WBC count in truncated group was lower significantly than that in the conventional group( P 0. 01). On the 4^(th) day of being infected,NLRP3 mRNA of mice lung tissue expressed highly in the model group,while decreased significan
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