黑骨藤皂苷对特应性皮炎小鼠的治疗作用及对Th1型免疫反应的影响  

Therapeutic effect of Periploca forrestii saponin on atopic dermatitis in mice and its influence on Th1 type immune response

作　　者：鲍春梅[1] 孙广臣[2] 

机构地区：[1]桂林医学院临床学院,广西桂林541000 [2]桂林医学院药学院

出　　处：《山东医药》2017年第13期32-35,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81460474;81260462)

摘　　要：目的探讨黑骨藤皂苷对特应性皮炎(AD)小鼠的治疗作用及对Th1型免疫反应的影响。方法 30只BLAB/c雌性小鼠随机分为正常组8只,模型组10只,黑骨藤皂苷组12只。除正常组外,其余两组均需造模。将小鼠腹部体毛剃去,用1.5%2,4-二硝基氯苯(DNCB)(丙酮∶橄榄油=4∶1)溶液100μL涂在腹部剃毛处初次致敏,2周后在小鼠耳廓内外两侧相同位置涂1%DNCB溶液20μL,1周重复刺激3次,连续2周,共造模30 d。从初次致敏起,黑骨藤皂苷组给予黑骨藤皂苷(0.002 g/d)灌胃,正常组和模型组分别给予等体积生理盐水灌胃,连续灌药30 d。从再次致敏当天(造模第14天)用游标卡尺测量每只小鼠两耳中部厚度,以后每次DNCB刺激前测量1次并记录,观察小鼠两耳外观肿胀情况。在末次耳部DNCB刺激12 h后取材,采用临床症状评分评价小鼠耳朵大体外观;切取耳部皮肤做病理切片,HE染色观察小鼠耳朵组织表皮厚度,甲苯胺蓝染色观察肥大细胞浸润情况;采用免疫组化法检测耳组织CD68蛋白的表达;采用ELISA法测定血清IgE、TNF-α、单核细胞趋化蛋白1(MCP-1)的表达。结果与模型组比较,不同时间正常组与黑骨藤皂苷组左耳厚度均降低(P<0.05或<0.01)。正常组、模型组、黑骨藤皂苷组临床症状评分组间两两比较,P均<0.01。正常组、模型组、黑骨藤皂苷组耳部组织表皮厚度组间两两比较,P均<0.01;真皮层肥大细胞浸润数目组间两两比较,P均<0.01。与正常组比较,模型组组织CD68及血清TNF-α、IgE、MCP-1水平均升高;与模型组比较,黑骨藤皂苷组组织CD68蛋白及血清TNF-α、IgE、MCP-1水平均降低(P<0.05或<0.01)。结论黑骨藤皂苷对AD小鼠有治疗作用,可抑制Th1型免疫反应。Objective To investigate the therapeutic effect of Periploca forrestii saponin（ PFS） on atopic dermatitis（ AD） in mice and its influence on Th1 type immune response. Methods Thirty female BLAB/c mice were randomly divided into the normal group（ n = 8）,model group（ n = 10）,and PFS group（ n = 12）. In addition to the normal group,mice in the other two groups were modeled. The mouse abdomen body was shaved and coated with 1. 5% 2,4-dinitrochlorobenzene（ DNCB）（ acetone∶ olive oil = 4∶ 1） solution 100 μL for the initial sensitization at the abdominal shaving,after 2weeks,20 μL of 1% DNCB solution was applied at the same position on both sides of the mouse auricle,we repeated stimulation 3 times in 1 week,and for 2 weeks,a total of 30 d. From the the initial immunization,the PFS group was given PFS（ 0. 002 g/d）,and the normal group and the model group were given equal volume of normal saline for 30 days. The thickness of the middle ears of each mouse was measured with a vernier caliper on the day of re-sensitization（ day 14）,and the mice were observed and recorded every time before the DNCB was stimulated to observe the swelling of the ears of the mice. At 12 h after last ear DNCB stimulation,the appearance of mouse ears was evaluated by clinical symptom score. The epidermis thickness of the ear tissues was observed by HE staining,and the infiltration of mast cells was observed by toluidine blue staining. The expression of CD68 protein was detected by immunohistochemistry. The levels of Ig E,TNF-α and MCP-1 in serum were measured by ELISA. Results Compared with the model group,the thickness of the left ear of the normal group and the PFS group decreased,P 〈 0. 05 or P 〈 0. 01. Significant differences were found in the clinical evaluations,ear skin tissue thickness and dermal layer mast cell infiltration number between the normal group,model group and PFS group,all P 〈 0. 01. Compared with the normal group,CD68 protein,serum TNF-α,Ig E and MCP-1 increased in the model gro

关 键 词：特应性皮炎 黑骨藤皂苷 Th1型免疫反应 CD68 IGE 肿瘤坏死因子 小鼠 

分 类 号：R758.2[医药卫生—皮肤病学与性病学]