摘要
目的通过检测鼠流感的肺病理变化及脾细胞Th1/Th2免疫因子及气管粘膜TLR2/4 mRNA的表达变化,初步探讨冰香散挥发油制剂对流感病毒的防治作用及其免疫调节机制。方法选用BALB/c小鼠,随机分为正常组、病毒对照组、利巴韦林组、冰香散挥发油制剂滴鼻组、雾化组,采用不同的给药方式预防性给药7d,第8天以15LD50攻毒,再持续给药5天后取材。肺组织经HE染色后观察其病变程度;脾脏研碎制得脾细胞悬浮液,以RT-PCR法检测细胞因子IFN-γ、IL-4、IL-5的分泌情况;RT-PCR检测气管粘膜上TLR2/4基因表达的水平。结果小鼠经H1N1感染后肺部出现严重的炎性细胞浸润,与正常组相比TLR2/4 mRNA、IFN-γmRNA表达水平上升(P<0.01),IL-4、L-5 mRNA表达水平下降(P<0.01)。与病毒对照组相比利巴韦林组可显著增加IL-4 mRNA的表达(P<0.01),降低IFN-γmRNA表达(P<0.05);冰香散挥发油制剂滴鼻、雾化组能显著增加IL-4 mRNA、IL-5 mRNA的表达水平(P<0.01或P<0.05),但在降低IFN-γmRNA的表达上冰香散挥发油制剂滴鼻组的效果比较显著(P<0.05)。各给药组都有降低TLR2/4基因表达的趋势,但只有冰香散挥发油制剂滴鼻组的作用比较显著(P<0.01)。结论冰香散挥发油制剂对流感病毒性肺炎病变有一定的防治作用,调控炎性细胞因子及TLR2/4基因的表达水平可能是其抗流感所致肺炎的重要作用机制之一。
Objective To explore the underlying mechanisms of Bing - Xiang Powder on influenza prevention by examining the histopathology of lungs and detecting the level of mRNA expression levels of Th1/Th2 cytokines and TLRs in influenza infected mice. Methods BALB/c mice of either sex were randomly divided into 5 groups, including Negative group, Influenza control group, Ribavirin group, Bing - Xiang Powder aerosol group, Bing - Xiang Powder intranasal group. All mice were intranasally chal- lenged with 15LD50 （ LD50 = 10^-5.5） doses of FM1 except the negative control group. Mice were treated with Bing - Xiang Powder （in aerosol or intranasal route） or ribavirin （oral administration） for 7 days before virus infection,followed by additional 5 days. The histopathology of lungs were stained with hematoxylin and eosin（HE） and examined under microscopy in a double - blinded manner. Single splenic cell suspensions were made by grinding the spleens and filtering through stainless steel . The mRNA expres- sion levels of IFN - γ, IL - 4, IL - 5 and the pathogen recognition receptors （ TLR2, TLR4 ） in the tracheal mucosa of infected mice were detected by use of RT - quantity PCR methods. Results The photomicrograph showed large area of pneumonia with intense exudates in bronchial lumen, severe alveolitis with marked infiltration of neutrophiles in the bronchiolar and the mRNA expression of TLR2 ,TLR4 in Influenza control group markedly higher than that in Negative group （P 〈 0.01 ） with a decreased level of IL -4, IL -5mRNA expression（ P 〈 0.01 ） . Bing - Xiang Powder（ in aerosol or intranasal route） promoted the expression level of IL - 4 mRNA, IL - 5mRNA（ P 〈 0.01 or P 〈 0.05 ）. However, the expression level of IFN - γmRNA in Bing - Xiang Powder intranasal group was lower than that of Bing - Xiang Powder aerosol group （ P 〈 0.05 ）. Bing - Xiang Powder intranasal group reduced the mRNA expression level of TLR2, TLR4 most dramatically among all of the groups （ P 〈 0.01 ）. Co
出处
《时珍国医国药》
CAS
CSCD
北大核心
2015年第4期797-800,共4页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(No.81072948)
广东省自然科学基金研究团队项目(No.S20120030006598)
广东省建设中医药强省立项资助科研课题(No.2010397)
