冬虫夏草菌丝体提取物调控免疫改善肾纤维化的实验研究  

An experimental study on effects of Hirsutella sinensis mycelium( HSM) extract on immunoregulation and kidney fibrosis

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3期354-357,共4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)小鼠的免疫反应和肾纤维化程度的影响。方法:建立CLP诱导脓毒症小鼠模型,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小时和术后10天给予HSM提取物200 mg/(kg·d),假术组和模型组给予等量的生理盐水。利用流式细胞术分析了小鼠外周血中巨噬细胞、中性粒细胞和Treg细胞数量以及腹水中巨噬细胞数量;利用QPCR检测了肾脏组织中炎性因子TNF-α和IL-1β及纤维化因子TGF-β1、TIMP1和MMP9的表达;另外,还对肾脏组织切片进行了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:模型组外周血中巨噬细胞、中性粒细胞和腹水中巨噬细胞数量较假术组显著升高,而HSM提取物治疗能下降上述细胞的数量;与正常组相比,模型组和治疗组外周血中Treg细胞数量均无明显差异;模型组肾脏组织中炎性因子TNF-α和IL-1β及纤维化因子TGF-β1、TIMP1和MMP9表达均上调,而治疗组肾脏上述细胞因子表达都有显著降低;HE染色显示模型组肾脏组织炎性样变明显,而治疗组有较好改善;与假术组比,模型组肾小管及间质α-SMA和fibronection表达明显增多,而治疗组肾小管及间质表达低于模型组。结论:HSM提取物具有调控免疫及改善肾纤维化的功能。Objective： To investigate the effect of HSM on the immunoreaction and renal fibrosis in mice with cecal ligation and puncture（ CLP）. Methods： Renal fibrosis was induced by CLP; mice were 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·d） ]was orally administered to HSM group2 hour before surgery and repeated everyday throughout 10 days,while sham group and model group were given the same dose of normal saline. FACS assay was used to analyze the amount of macrophages,neutrophils and Treg in PBMC,as well as macrophages in peritoneal fluid; 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 renal sections. Besides,renal sections were subjected to HE stain and immunohistochemical staining with α-SMA and fibronectin. Results： The amount of model group＇s macrophages and neutrophils in PBMC,as well as macrophages in the peritoneal fluid were significantly higher than sham group＇ s,whereas HSM succeeded in lowering them; contrast to sham group,Tregs＇ amount of CLP group and HSM group in PBMC had no significant changes. The expression of inflammatory molecules（ IL-1β and TNF-α） and fibrogenic cytokines（ TGF-β,MMP9 and TIMP1） from CLP group＇s renal sections were remarkably improved,whereas HSM inhibited that. The CLP group＇s HE results showed obvious renal inflammatory damage,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 could regulate immunity response and had effect in improving renal fibrosis.

关 键 词：冬虫夏草菌丝体 盲肠穿孔结扎 免疫反应 纤维化 

分 类 号：R392[医药卫生—免疫学]