冬虫夏草菌丝体提取物通过抑制炎性因子及Treg细胞功能缓解肺纤维化  

Hirsutella sinensis mycelium(HSM) extract relieves pulmonary fibrosis though suppress inflammation and function of Treg cells

作　　者：岳会敏[1] 刘飞[1] 李范林 马菲雅 沈苏南[1] 侯亚义[1] 

机构地区：[1]南京大学医学院和生物医药技术国家重点实验室,南京210093

出　　处：《中国免疫学杂志》2016年第10期1472-1476,共5页Chinese Journal of Immunology

摘　　要：目的:观察冬虫夏草菌丝体(Hirsutella sinensis mycelium,HSM)提取物对盲肠穿孔结扎(CLP)小鼠的炎症反应和肺纤维化程度的影响。方法:建立CLP诱导脓毒症5 d和10 d小鼠模型,分别分为假术组(sham,n=5)、模型组(CLP,n=11)、治疗组(HSM,n=11)。冬虫夏草菌丝体组在术前2 h和术后给予HSM提取物200 mg/(kg·d)(低于人体用量0.27 g/kg),假术组和模型组给予等量的生理盐水。利用Q-PCR检测了肺组织中炎性因子TNF-α和IL-1β及纤维化因子TGF-β1、TIMP1和MMP9的表达;利用流式细胞术分析了小鼠外周血和脾脏中Th1细胞以及脾脏组织中Treg细胞的数量;另外,还对肺组织切片进行了HE染色以及α-平滑肌肌动蛋白(α-SMA)和纤连蛋白(fibronection)免疫组化染色。结果:模型组外周血和脾脏中Th1细胞以及脾脏组织中Treg细胞的数量较假术组显著降低,而HSM提取物治疗能上调上述细胞的数量。5 d时,模型组肺组织中IL-1β及纤维化因子TGF-β1、MMP9和TIMP1表达显著上调,到第10天时有所降低但仍高于sham组,而治疗组上述因子表达明显降低;各组肺组织TNF-α表达无显著差异;HE染色显示模型组肺部炎性样变明显,到第10天时炎性样变比5 d时减轻,而治疗组均有改善;免疫组化显示与假术组比,模型组肺内间质α-SMA和fibronection表达明显增多,而治疗组肺内间质表达低于模型组。结论:HSM提取物具有抑制炎症、平衡免疫及缓解肺纤维化的功能。Objective： To investigate the effect of HSM on the inflammation and pulmonary fibrosis in mice with cecal ligation and puncture（ CLP）. Methods： Both 5 day and 10 day timepoint pulmonary fibrosis were induced by CLP; mice were randomly divided into three groups,including sham group（ sham,n = 5）,model group（ CLP,n = 11）,therapy group（ HSM,n = 11）. HSM extract（ 200 mg/kg,less than human dose of 0. 27 g/kg） was orally administered to HSM group 2 hour before surgery and repeated everyday,while sham group and model group were given the same dose of normal saline. We used Q-PCR assay to analyze the expression of inflammatory molecules（ IL-1β and TNF-α） and fibrogenic cytokines（ TGF-β,MMP9 and TIMP1） from lung tissues,we used FACS assay to analyze the amount of Th1 cells in PBMC and spleen sections,as well as Treg in spleen sections. Besides,lung sections were subjected to HE stain and immunohistochemical staining with α-SMA and fibronectin. Results： The amount of model group＇s Th1 cells in PBMC and spleen sections,as well as Treg in spleen sections were significantly lower than sham group＇ s,whereas HSM succeeded in raising them. By day 5,the expression of inflammatory molecules IL-1β and fibrogenic cytokines（ TGF-β,MMP9 and TIMP1） from CLP group＇s lung sections were remarkably improved,by day 10,the expression had decreased but was higher than sham group＇s; HSM inhibited these cytokines＇ expression. The expression of TNF-α of CLP group and HSM group had no significant changes. The CLP group＇s HE results showed obvious pulmonary inflammatory damage,by day 10,the situation had improved,whereas HSM reduced the histopathologic alterations; contrast to sham group,model group＇s expression of α-SMA and fibronectin was remarkably improved,while HSM group showed lower expression. Conclusion： HSM extract can suppress inflammation,balances immune system and relieves pulmonary fibrosis.

关 键 词：冬虫夏草菌丝体 炎症反应 肺纤维化 

分 类 号：R392[医药卫生—免疫学]