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目的研究血吸虫可溶性虫卵抗原(SEA)与哮喘发生的关系及其对CD4+CD25+调节性T细胞(CD4+CD25+Treg)及相关基因Foxp3表达的影响。方法 BALB/C小鼠腹腔及足垫共注射SEA 50μg/只,每周1次,共4次。对照组注射生理盐水。然后用卵清白蛋白(OVA)诱导小鼠产生过敏性哮喘。剖杀小鼠,取肺组织,做病理学检查;收集支气管肺泡灌洗液(BALF),涂片、染色后进行细胞分类计数;分离脾细胞,用流式细胞仪检测CD4+CD25+调节性T细胞(CD4+CD25+Treg)占CD4+T细胞的比例;提取脾脏RNA,以逆转录得到的cDNA为模板扩增Foxp3基因,检测脾脏Foxp3mRNA表达水平。结果 SEA免疫并OVA致哮喘组小鼠肺组织炎症较OVA单纯哮喘对照组轻,只有少量炎性细胞浸润。BALF涂片染色检查SEA免疫组BALF中的细胞密度低于对照组,其中嗜酸粒细胞占总细胞数的(2.22±1.52)%,对照组占(19.93±4.08)%,差异有统计学意义(P<0.05);流式细胞仪检测SEA免疫组小鼠脾细胞中CD4+CD25+Treg占CD4+T细胞总数的(32.24±2.19)%,对照组占(27.41±2.87)%,差异有统计学意义(P<0.05);PCR检测SEA免疫组脾细胞Foxp3mRNA表达水平高于对照组。结论 SEA对哮喘的发生有一定抑制作用,可能通过CD4+CD25+Treg影响机体的免疫调节机制。
Objective To study the relationship between schistosoma soluble egg antigen (SEA) and asthma and its effect on the expression of CD4 + CD25 + regulatory T cells (CD4 + CD25 + Treg) and related genes Foxp3. Methods BALB / C mice were injected intraperitoneally with footpads 50 μg / SEA once a week for 4 times. The control group was injected with saline. Mice were then challenged with ovalbumin (OVA) to induce allergic asthma. The mice were sacrificed and the lung tissues were removed for histopathological examination. Bronchoalveolar lavage fluid (BALF) was collected and stained. The cells were sorted and counted. The spleen cells were isolated and the percentage of CD4 + CD25 + regulatory T cells was measured by flow cytometry (CD4 + CD25 + Treg) accounted for the proportion of CD4 + T cells; spleen RNA was extracted, reverse transcription of the cDNA obtained as a template for amplification of Foxp3 gene, detection of spleen Foxp3 mRNA expression levels. Results SEA immunization and OVA-induced asthma in mice lung tissue inflammation than OVA simple asthma control group, with only a small amount of inflammatory cell infiltration. The BALF smear staining showed that the cell density of BALF in SEA-immunized group was lower than that in control group (2.22 ± 1.52%) and control group (19.93 ± 4.08%), the difference was statistically significant (32.24 ± 2.19)% of the total number of CD4 + T cells in the spleen cells of the SEA immunized group, and (27.41 ± 2.87)% in the control group, the difference was (P <0.05) Statistical significance (P <0.05). The level of Foxp3 mRNA in spleen cells of SEA immunized group was higher than that of control group by PCR. Conclusions SEA can inhibit the occurrence of asthma and may affect the immune regulation mechanism by CD4 + CD25 + Treg.