银翘散对呼吸道合胞病毒感染小鼠呼吸系统的作用研究
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R373.1

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Yinqiao Powder Has Effect on Respiratory System of Mice Infected with Respiratory Syncytial Virus
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    摘要:

    目的:探讨银翘散对呼吸道合胞病毒感染小鼠呼吸系统的治疗效果以及作用机制。方法:小鼠随机分为6组,每组9只,分别是正常组、模型组、银翘散高剂量组、银翘散中剂量组、银翘散低剂量组、利巴韦林组。除正常组外,其他各组小鼠用乙醚浅麻后,以浓度TCID50呼吸道合胞病毒(RSV)液50 μL滴鼻感染小鼠。银翘散高、中、低剂量组分别给予600、300、150 mg/(kg·d) 银翘散灌胃,利巴韦林组给予150 mg/(kg·d) 利巴韦林灌胃。银翘散高、中、低剂量组、利巴韦林组于感染前1天开始灌胃给药,连续5天,正常组、模型组给予等容积蒸馏水灌胃。通过称量小鼠体质量和肺重量计算肺指数。HE染色检测小鼠鼻黏膜和肺组织炎性细胞浸润等病理形态学的改变。酶联免疫吸附法测定检测小鼠肺泡灌洗液中白细胞介素-1β(IL-1β)、肿瘤坏死因子α(TNF-α) 以及白细胞介素-6(IL-6) 的表达水平。实时荧光定量PCR和Western blot检测肺组织核苷酸结合寡聚化结构域样受体3(NALP3) 炎性体、凋亡相关微粒蛋白(ASC) 和半胱天冬酶-1(caspase-1) 的表达。结果:与正常组比较,模型组小鼠肺指数显著升高,肺泡灌洗液中TNF-α、IL-1β和IL-6表达显著升高,肺组织NALP3、ASC、caspase-1 mRNA和蛋白表达显著升高,差异均有统计学意义(P<0.05)。与模型组比较,银翘散高、中、低剂量组、利巴韦林组小鼠肺指数显著降低,肺泡灌洗液中TNF-α、IL-1β、IL-6表达显著降低,肺组织NALP3、ASC、caspase-1 mRNA和蛋白表达显著降低,差异均有统计学意义(P<0.05)。与利巴韦林组比较,银翘散中剂量组小鼠肺泡灌洗液中TNF-α、IL-1β表达较高,肺组织NALP3和caspase-1 mRNA表达较高;银翘散低剂量组小鼠肺指数较高,肺泡灌洗液中TNF-α、IL-1β、IL-6表达较高,肺组织NALP3、ASC、caspase-1 mRNA和蛋白表达较高;差异均有统计学意义(P<0.05)。结论:银翘散能够改善呼吸道合胞病毒感染所致的呼吸道系统炎症,其作用机制与抑制NALP3炎性体的激活和炎症因子的分泌相关。

    Abstract:

    Objective:To discuss the clinical effect of Yinqiao powder on respiratory system of mice model infected with respiratory syncytial virus and its mechanism.Methods: Mice were divided into 6 groups randomly,namely the normal group,the model group,the group of low-dose Yinqiao powder,the group of medium-dose Yinqiao powder,the group of high-dose Yinqiao powder and the ribavirin group,9 mice in each group.Except the normal group,mice in other groups were infected with 50μL nasal drops of TCID50 respiratory syncytial virus(RSV)fluid after received light anesthesia with ether.The group of low-dose Yinqiao powder,the group of medium-dose Yinqiao powder and the group of high-dose Yinqiao powder were respectively given 600 mg/(kg·d),300 mg/(kg·d)and 150 mg/(kg·d)of Yinqiao powder by gavage,while the ribavirin group was given 150 mg/(kg·d)of ribavirin by gavage.The group of low-dose Yinqiao powder, the group of medium-dose Yinqiao powder,the group of high-dose Yinqiao powder and the ribavirin group began to receive medication by gavage one day before infection,continuously lasting for 5 days,while the normal group and the model group were given equal volume of distilled water by gavage.Lung indexes were calculated by weighing body mass and lung weight of mice.HE staining was used to detect the pathological changes in the nasal mucosa and inflammatory cells infiltration in the lung tissueof mice.The levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in alveolar lavage fluid were detected by enzyme-linked immunosorbent assay.The expressions of NALP3 inflammasome,apoptosis-associated microparticles(ASC)and caspase-1 in lung tissue were detected by real-time quantitative PCR and Western blot.Results:Compared with those in the normal group,the lung indexes of the model group were increased significantly,the expression of TNF-α,IL-1β,and IL-6 in alveolar lavage fluid was increased evidently,and the expression of NALP3,ASC,caspase-1 mRNA and protein in lung tissue were obviously increased,differences being significant(P<0.05).Compared with those in the model group,the lung indexes of mice in the groups of Yinqiao powder and the ribavirin group were evidently decreased, the expression of TNF-α, IL-1β, and IL-6 in alveolar lavage fluid was significantly decreased, and the expression of NALP3,ASC, caspase-1 mRNA and protein in lung tissue was obviously decreased, differences being significant(P<0.05).Compared with those in the ribavirin group,the expression of TNF-α and IL-1β in the alveolar lavage fluid of mice in the group of medium-dose Yinqiao powder was higher,and the expression of NALP3 and caspase-1 mRNA in lung tissue were higher, while the lung indexes of mice in the group of low-dose Yinqiao powder were higher, the expression of TNF-α,IL-1β and IL-6 in the alveolar lavage fluid was higher,and the expression of NALP3,ASC,caspase-1 mRNA and protein in lung tissue were higher,differences being significant(P<0.05).Conclusion:Yinqiao powder can improve respiratory system inflammation resulting from infection of respiratory syncytial virus,and its mechanism is related to the inhibition of the activation of NALP3 inflammasome and the secretion of inflammatory cytokines.

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覃黎葵,张奉学,黄笑娟,宋鸿,龚文亮.银翘散对呼吸道合胞病毒感染小鼠呼吸系统的作用研究[J].新中医,2018,50(5):24-30

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  • 在线发布日期: 2018-05-03
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