摘要
目的探讨PU.1抑制剂DB2313对MRL/lpr小鼠狼疮性肾炎(lupus nephritis,LN)的改善作用及机制。方法将30只雌性MRL/lpr小鼠随机分配至3组:模型组、DB2313组、泰他西普(TACI-Ig)组,每组10只。选取另外10只雌性BALB/c小鼠作为对照组。DB2313组小鼠隔日腹腔DB2313注射,TACI-Ig组隔日皮下注射TACI-Ig。每日腹腔注射给予对照组与模型组等量0.9%NaCl注射液。在药物干预前及干预后的1~5周内,定期收集小鼠尿液,测定尿蛋白含量,评估肾脏损伤指数。HE、Masson及PAS染色观察肾组织病理学变化;免疫组化方法检测肾组织内免疫复合物C3含量;试剂盒检测血清中肌酐(Scr)、尿素氮(BUN)、IL-6及TNF-α的水平;免疫荧光技术测定肾组织中PU.1与FLT3的表达水平;Western blot技术检测肾组织PU.1、FLT3、PI3K、AKT及p-AKT的蛋白表达。结果DB2313治疗能明显减轻MRL/lpr小鼠肾脏病理损伤,降低C3沉积、肾脏损伤指数及24 h尿蛋白量;ELISA结果表明,DB2313给药可明显降低MRL/lpr小鼠血清BUN、Scr、IL-6及TNF-α的水平;免疫荧光与Western blot结果进一步表明,DB2313治疗可明显下调PU.1、PI3K及p-AKT的蛋白表达,上调FLT3的蛋白表达。结论DB2313对MRL/lpr小鼠LN具有改善作用,其潜在机制可能涉及抑制转录因子PU.1介导的信号通路。
Aim To investigate the effect of PU.1 inhibitor DB2313 on lupus nephritis in MRL/lpr mice and its mechanism.Methods Thirty female MRL/lpr mice were randomly divided into the model group,DB2313 group and TACI-Ig group,with 10 mice in each group.Another 10 female BALB/c mice were selected as normal control groups.Mice in the DB2313 group received intraperitoneal DB2313 injections every two days,and those in the TACI-Ig group received subcutaneous injections of TACI-Ig every two days.Mice in the control group and model group were intragastrically given the same amount of 0.9%NaCl injection every day.Before the drug intervention and for 1 to 5 weeks after the intervention,the urine of mice was collected regularly,the urine protein content was measured,and the renal damage index was evaluated.The histopathological changes of kidney were observed by HE,Masson and PAS staining.The expression levels of immune complex of C3 in kidney tissue were detected by immunohistochemistry.The concentrations of urea nitrogen(BUN),serum creatinine(Scr),interleukin-6(IL-6),and tumor necrosis factor alpha(TNF-α)in the serum samples were assayed utilizing the respective kits.The expression levels of PU.1 and FLT3 in kidney tissues were determined by immunofluorescence technology,and the protein expressions of PU.1,FLT3,PI3K,AKT and phosphorylated AKT(p-AKT)in kidney tissues were detected by Western blot.Results DB2313 treatment significantly alleviated the pathological damage of kidney in MRL/lpr mice,and reduced the deposition of C3,kidney injury index and 24-hour urine protein in renal tissue.The results of ELISA showed that DB2313 administration could significantly reduce the serum levels of BUN,Scr,IL-6 and TNF-αin MRL/lpr mice.The results of immunofluorescence and Western blot further showed that DB2313 treatment could significantly down-regulate the protein expression of PU.1,PI3K and p-AKT,and up-regulate the protein expression of FLT3.Conclusion DB2313 has an ameliorating effect on lupus nephritis in MRL/lpr mice,and its underlying mechanism may involve the inhibition of the transcription factor PU.1-mediated signaling pathway.
作者
徐诺
郭婷婷
李颖
王康
魏伟
严尚学
XU Nuo;GUO Ting-ting;LI Ying;WANG Kang;WEI Wei;YAN Shang-xue(Institute of Clinical Pharmacology,Key Laboratory of Anti-inflammatoryand Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-infammatory and Immuno Drugs,Anhui Medical University;Experimental Animal Center,Anhui Medical University,Hefei 230032,China)
出处
《中国药理学通报》
北大核心
2025年第8期1478-1484,共7页
Chinese Pharmacological Bulletin
基金
安徽高校自然科学研究重大项目(No KJ2020ZD15)
安徽省转化医学研究院科研基金重点项目(No 2023zhyx-B14)。
作者简介
徐诺(2000-),女,硕士生,研究方向:抗炎免疫药理学,E-mail:2529649350@qq.com;通信作者:严尚学(1973-2025),男,研究员,研究方向:抗炎免疫药理学,硕士生导师,E-mail:yan-shx@163.com。
