BAFF对免疫性血小板减少症模型小鼠的氧化应激和炎症反应的调控机制  

Study on the regulatory mechanism of BAFF on oxidative stress and inflammatory response in a mouse model of immune thrombocytopenia

作　　者：李巍[1] 马西虎[1] 刘晓[1] 费飞 秦兰[1] 买尔吾甫·木合布力 白玉盛[1] LI Wei;MA Xihu;LIU Xiao;FEI Fei;QIN Lan;MAIERWUFU Muhebuli;BAI Yusheng(Department of Hematology,The Fourth Affiliated Traditional Chinese Medicine Hospital of Xinjiang Medical University,Urumqi 830001,China;Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第四附属中医医院血液科,新疆乌鲁木齐830001 [2]新疆医科大学,新疆乌鲁木齐830054

出　　处：《西部医学》2025年第2期168-174,179,共8页Medical Journal of West China

基　　金：新疆维吾尔自治区自然科学基金项目(2022D01C167)。

摘　　要：目的 探讨B细胞激活因子(BAFF)对免疫性血小板减少症模型小鼠的氧化应激和炎症反应的影响和潜在机制。方法 制备豚鼠抗小鼠血小板抗血清(GP-APS),并将50只无特定病原级别的成年雄性BALB/c小鼠(7~8周龄)分为对照组(空白对照)、ITP组(GP-APS诱导)、ITP+rhBAFF组(ITP组联合静脉注射50μg/kg/50μL重组人BAFF蛋白)、ITP+rhBAFF+shDll1(100μg/kg)组和ITP+rhBAFF+shJagged1(25μg/kg)组[构建了沉默Dll1的重组腺病毒载体(shDll1)和沉默Jagged1的重组腺病毒载体(shJagged1),在ITP+rhBAFF组的基础上静脉注射腺病毒载体,每日1次,注射1周],每组10只。1周后取小鼠外周血,并分离单个核细胞和血清。对外周血中的血小板进行计数。用酶联免疫吸附法(ELSIA)检测小鼠外周血血清中BAFF、Dll1、Jagged1、白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-α、活性氧(ROS)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-PX)、超氧化物歧化酶(SOD)的水平。皮尔森相关性分析法分析外周血血清中BAFF分别与Dll1和Jagged1的相关性。Western blot检测小鼠外周血单个核细胞中BAFF、Dll1、Jagged1的蛋白表达。结果 与对照组相比,ITP组小鼠的外周血血小板数目降低(P<0.05)。ITP小鼠外周血血小板数与BAFF的水平呈显著负相关(r=-0.5687;P=0.0338,95%CI:-0.8445~-0.05464);外周血血小板数与Dll1的水平呈显著负相关(r=-0.6361;P=0.0145,95%CI:-0.8723~-0.1592);外周血血小板数与Jagged1的水平呈显著负相关(r=-0.6409;P=0.0135,95%CI:-0.8742~-0.1672)。ITP小鼠外周血中BAFF的水平与Dll1呈显著正相关(r=0.5794;P=0.0236,95%CI:0.09544~0.8418)。BAFF的水平与Jagged1的水平呈显著性正相关(r=0.5723;P=0.0258,95%CI:0.08489~0.8387)。与对照组比较,ITP组外周血中BAFF、Dll1、Jagged1、IL-1β、IL-6、TNF-α、ROS、MDA水平上调,GSH-PX、SOD水平下调(均P<0.05)。与ITP组比较,ITP+rhBAFF组外周血中BAFF、Dll1、Jagged1、IL-1β、IL-6、TNF-α、ROS、MDA水平上调,�Objective To investigate the effects and potential mechanisms of B-cell activating factor(BAFF) on oxidative stress and inflammatory response in a mouse model of immune thrombocytopenia(ITP).Methods Guinea pig anti-mouse platelet antiserum(GP-APS) was prepared,and 50 specific pathogen-free adult male BALB/c mice(7-8 weeks old) were randomly divided into 5 groups with n=10 in each group.The groups included a control group(blank control),an ITP group(induced by GP-APS),an ITP+rhBAFF group(ITP combined with intravenous injection of 50 μg/kg/50 μL recombinant human BAFF protein).Recombinant adenovirus vectors silencing Dll1(shDll1) and Jagged1(shJagged1) were constructed,and adenovirus vectors were intravenously injected on top of the ITP+rhBAFF group once a day for 1 week,named the ITP+rhBAFF+shDll1(100 μg/kg) group and ITP+rhBAFF+shJagged1(25 μg/kg) group,respectively.After 1 week,peripheral blood was collected from the mice and peripheral blood mononuclear cells and serum were separated.Platelet counts in peripheral blood were measured.Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of BAFF,Dll1,Jagged1,interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-α,reactive oxygen species(ROS),malondialdehyde(MDA),glutathione peroxidase(GSH-PX),and superoxide dismutase(SOD) in mouse serum.Pearson correlation analysis was used to analyze the correlation between BAFF and Dll1,Jagged1 levels in peripheral blood serum.Western blot was performed to detect the protein expression of BAFF,Dll1,and Jagged1 in peripheral blood mononuclear cells.Results Compared to the control group,the peripheral blood platelet count in the ITP group of mice was significantly reduced(P<0.05).There was a significant negative correlation between the peripheral blood platelet count in ITP mice and the level of BAFF(r=-0.5687;P=0.0338,95% CI:-0.8445 to-0.05464).Similarly,a significant negative correlation was observed between the peripheral blood platelet count and the level of Dll1(r=-0.6361;P=0.0145,95% CI:-0.8723 to-0.1592

关 键 词：B细胞激活因子 免疫性血小板减少症 小鼠 氧化应激 炎症反应 

分 类 号：R558.2[医药卫生—血液循环系统疾病]