摘要
目的探讨维生素D对肥胖型哮喘模型小鼠高迁移率组蛋白B1/高糖基化终末产物受体(HMGB1/RAGE)通路和脂肪因子水平的调节作用。方法实验完成于2023年2—9月,实验平台位于嘉兴学院第二医院实验中心及嘉兴学院实验室。30只小鼠采用数字耳号进行标识,并通过耳号数字随机分为Ⅰ组(正常对照组)、Ⅱ组(哮喘组)、Ⅲ组(肥胖型哮喘组)、Ⅳ组(哮喘+维生素D组)和Ⅴ组(肥胖型哮喘+维生素D组),每组6只。单纯肥胖小鼠模型通过高脂饮食诱导,哮喘模型小鼠需采用卵清蛋白(OVA)腹腔注射致敏和雾化吸入方式诱导,维生素D干预模型小鼠则按照1 mL/d连续灌胃2周方式完成干预。采用酶联免疫吸附试验(ELISA)法检测小鼠血液中白细胞介素4(IL-4)、IL-6、IL-10、脂联素及瘦素等因子水平,使用定量逆转录聚合酶链反应(qRT-PCR)方法检测HMGB1和RAGE基因的表达,最后对上述结果采用SPSS进行统计学分析。结果Ⅱ组支气管肺泡灌洗液(BALF)中白细胞计数(1.34±0.48)×10^(5)/L,与Ⅳ组的(4.07±0.14)×10^(5)/L比较,差异有统计学意义(t=-18.28,P<0.001);Ⅲ组BALF中白细胞计数(9.61±0.91)×10^(5)/L,与Ⅴ组的(4.89±0.38)×10^(5)/L比较,差异有统计学意义(t=11.75,P<0.001)。Ⅱ组BALF中的嗜酸性粒细胞百分比(28.75±1.94)%,与Ⅳ组的(11.51±1.99)%比较,差异有统计学意义(t=15.20,P<0.001);Ⅴ组BALF中嗜酸性粒细胞百分比为(12.50±1.42)%,与Ⅲ组的(29.80±1.96)%比较,差异有统计学意义(t=17.74,P<0.001)。ELISA结果显示,与Ⅲ组相比,Ⅴ组血清IL-4、IL-6、白介素17(IL-17)、白介素1β(IL-1β)、免疫球蛋白E(IgE)、肿瘤坏死因子α(TNFα)水平均显著降低,两组差异均有统计学意义(t=15.24、9.65、2.26、5.83、10.86、2.50,均P<0.001);Ⅲ组血清IL-10表达水平为(4.97±0.25)pg/mL,与Ⅴ组的(8.84±0.64)pg/mL比较,差异有统计学意义(t=-13.89,P<0.001);Ⅴ组血清脂联素水平为(1.95±0.85)mg/L,与Ⅲ组的(1.15±0.13)
Objective To investigate the regulatory effect of vitamin D on the HMGB1/RAGE pathway and adipokine levels in a mouse model of obesity and asthma.Methods This study was conducted at the Experimental Center of the Second Affiliated Hospital of Jiaxing University and the laboratory of Jiaxing University from February to September 2023.Thirty mice were marked with digital ear numbers and were randomly divided into five groups,with six mice in each group:GroupⅠ(normal control group),GroupⅡ(asthma group),GroupⅢ(obesity and asthma group),GroupⅣ(asthma+vitamin D group),and Group V(obesity and asthma+vitamin D group).An obesity mouse model was induced using a high-fat diet,while an asthma mouse model was induced through sensitization via intraperitoneal injection of ovalbumin and aerosol inhalation.The vitamin D intervention consisted of continuous intragastric administration of vitamin D(1 mL/d)for 2 weeks.Blood levels of interleukin-4,interleukin-6,interleukin-10,adiponectin,and leptin were determined using the enzyme-linked immunosorbent assay method.The expression of genes encoding high mobility group protein B1(HMGB1)and the receptor for advanced glycation end products(RAGE)was detected using a quantitative reverse transcription polymerase chain reaction.All the obtained results were statistically analyzed using SPSS software.Results The white blood cell count in bronchoalveolar lavage fluid(BALF)of GroupⅡwas(1.34±0.48)×10^(5)/L,which was significantly lower than(4.07±0.14)×10^(5)/L in GroupⅣ(t=-18.28,P<0.001).The white blood cell count in BALF in GroupⅢwas(9.61±0.91)×10^(5)/L,which was significantly higher than(4.89±0.38)×10^(5)/L in Group V(t=11.75,P<0.001).The percentage of eosinophils in BALF of Group II was(28.75±1.94)%,which was significantly higher than(11.51±1.99)%in GroupⅣ(t=15.20,P<0.001).The percentage of eosinophils in BALF of Group V was(12.50±1.42)%,which was significantly lower than(29.80±1.96)%in GroupⅢ(t=17.74,P<0.001).The ELISA results demonstrated that serum levels of
作者
范宜佳
祝峰
朱晓凤
Fan Yijia;Zhu Feng;Zhu Xiaofeng(Department of Pediatrics,Second Affiliated Hospital of Jiaxing University,Jiaxing 314000,Zhejiang Province,China;Department of Orthopedics,The Second Hospital of Jiaxing,Jiaxing 314000,Zhejiang Province,China)
出处
《中国基层医药》
CAS
2024年第7期970-975,共6页
Chinese Journal of Primary Medicine and Pharmacy
基金
浙江省嘉兴市科技计划(2022AD30023)。
