CAJ | 학술논문

目的探讨药理剂量维生素C(5 ～20 mmol/L)体内抗流感病毒A/CA/7/09(H1N12009)的作用及效果.方法 BALB/c小鼠感染流感病毒A/CA/7/09后接受维生素C溶液皮下注射或经口灌胃,对照组为等量生理盐水,每组4～6只小鼠,每天2次,共14 d.每天监测小鼠的体重和死亡情况.在不同时间点取小鼠肺组织,用半数组织培养感染剂量法(TCID50)测定病毒滴度;测定肺组织中炎性因子IL-1β、IL-6、肿瘤坏死因子-α(TNF-α)和干扰素-α(IFN-α)含量;将肺组织切片染色,显微镜下观察炎症反应情况并进行病理评分.结果小鼠感染流感病毒后分别接受生理盐水和药理剂量维生素C治疗,第9天开始生理盐水组出现小鼠死亡,2周内有10％小鼠死亡或体重下降超过25％.而药理剂量维生素C治疗组2周内无小鼠死亡或体重下降超过25％.药理剂量维生素C治疗组小鼠肺内病毒滴度下降至对照组的1/10～1/100,且肺内病毒更早被清除.感染后第4天,药理剂量维生素C治疗组和对照组的肺组织中IL-1含量分别为(0.41±0.13)和(1.06±0.27) ng/g(t =9.36,P＜0.05),IL-6含量分别为(2.17±0.47)和(1.07±0.16) ng/g(t=12.2,P＜0.05),IFN-α含量分别为(1.52±0.3)和(0.84 ±0.19) ng/g(=6.82,P＜0.05).肺组织炎症反应(包括坏死性支气管炎、血管周围炎、细支气管周围炎和肺泡炎等)较对照组轻.结论药理剂量维生素C能降低肺部流感病毒的滴度,降低肺部炎症反应及感染老鼠的死亡.
목적 탐토약리제량유생소C(5 ～20 mmol/L)체내항류감병독A/CA/7/09(H1N12009)적작용급효과.방법 BALB/c소서감염류감병독A/CA/7/09후접수유생소C용액피하주사혹경구관위,대조조위등량생리염수,매조4～6지소서,매천2차,공14 d.매천감측소서적체중화사망정황.재불동시간점취소서폐조직,용반수조직배양감염제량법(TCID50)측정병독적도;측정폐조직중염성인자IL-1β、IL-6、종류배사인자-α(TNF-α)화간우소-α(IFN-α)함량;장폐조직절편염색,현미경하관찰염증반응정황병진행병리평분.결과 소서감염류감병독후분별접수생리염수화약리제량유생소C치료,제9천개시생리염수조출현소서사망,2주내유10％소서사망혹체중하강초과25％.이약리제량유생소C치료조2주내무소서사망혹체중하강초과25％.약리제량유생소C치료조소서폐내병독적도하강지대조조적1/10～1/100,차폐내병독경조피청제.감염후제4천,약리제량유생소C치료조화대조조적폐조직중IL-1함량분별위(0.41±0.13)화(1.06±0.27) ng/g(t =9.36,P＜0.05),IL-6함량분별위(2.17±0.47)화(1.07±0.16) ng/g(t=12.2,P＜0.05),IFN-α함량분별위(1.52±0.3)화(0.84 ±0.19) ng/g(=6.82,P＜0.05).폐조직염증반응(포괄배사성지기관염、혈관주위염、세지기관주위염화폐포염등)교대조조경.결론 약리제량유생소C능강저폐부류감병독적적도,강저폐부염증반응급감염로서적사망.
Objective To investigate the effects of pharmacologic ascorbate (vitamin C) against Influenza A/CA/7/09 (H1N12009).Methods BALB/c mice inoculated intranasally with influenza virus were treated with ascorbate (3 mg/g) twice daily by intraperitoneal (i.p.) injection for up to 14 d.Control groups received an equivalent volume of normal saline.Body weights were measured daily.To quantify the level of viral replication in the respiratory tract,the mice were euthanized and lungs removed and prepared as whole lung homogenates.Viral titers were determined by TCID50 assay in MDCK cells.Cytokine titers were determined by ELISA following the manufacturer' s protocol (IL-1 β,IL-6,TNF-α,IFN-α).For lung histopathologic evaluation,lungs were fixed with 10％ neutral-buffered formalin at time of isolation,and then coded,processed into paraffin blocks,sectioned onto glass slides and stained (hematoxylin and eosin).Slides were examined and scored by a pathologist.Results Mice infected with influenza virus and treated with pharmacologic ascorbate had higher survival and less weight loss,and had lung viral titers reduced by as much as 10 to 100-fold compared to the controls.Pathologic study of the lungs showed that the treated animals had little inflammation (bronchiolitis,perivasculitis,alveolitis,and vasculitis) compared to the controls.IL-1,IL-6,and IFN-alpha lung levels were lower in the treated animals compared to the controls.Conclusion Pharmacologic ascorbate is a pro-oxidant that eliminates influenza virus in the lung,and therefore reduces lung inflammation and lowers death rate in this mouse model.