Sigirr缺失调控NF-κB并参与慢性肾病小鼠发生肾间质纤维化  

Sigirr deletion upregulates NF-κB and is involved in the development of renal interstitial fibrosis in mice with chronic kidney disease

作　　者：童子文 徐德苹 王喆 杨萍 涂珍珍[1] 臧丹丹 周海胜[1,3] Tong Ziwen;Xu Deping;Wang Zhe;Yang Ping;Tu Zhenzhen;Zang Dandan;Zhou Haisheng(Dept of Biochemistry,Anhui Medical University,Hefei 230032;School of Basic Medical Science,Anhui Medical University,Hefei 230032;Center for Scientific Research,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学生物化学教研室,合肥230032 [2]安徽医科大学基础医学院,合肥230032 [3]安徽医科大学科研实验中心,合肥230032

出　　处：《安徽医科大学学报》2023年第5期724-730,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82071832)。

摘　　要：目的研究Sigirr基因缺失小鼠在慢性肾病(CKD)并发肾间质纤维化(RIF)中的作用和机制。方法利用PCR鉴定正常基因型(Sigirr^(+/+))小鼠和Sigirr基因缺失(Sigirr^(-/-))小鼠。含0.2%高腺嘌呤饲料喂养小鼠,连续喂养12周,以建立慢性肾脏损伤并发RIF模型。收集小鼠眼框静脉血以检测肾功能;利用HE染色分析肾脏组织病理变化,通过Masson染色观察肾脏纤维化程度,利用IHC和Western blot检测白细胞介素(IL)-1β、MyD88和NF-κB等信号分子变化,同时观察转化生长因子(TGF)-β1、E-cadherin和Vimentin的表达变化。结果肾功能检测结果显示高腺嘌呤喂养小鼠的血清肌酐和尿素氮较对照组增加;HE和Masson染色结果显示:与Sigirr^(+/+)小鼠比较,Sigirr^(-/-)小鼠的肾组织中,炎症细胞浸润和胶原纤维沉积更明显。IHC和Western blot结果显示IL-1β及其下游的MyD88表达增加,p-P65水平显著增加;Sigirr^(-/-)小鼠的肾组织中IL-1β、MyD88、p-P65等变化较Sigirr^(+/+)小鼠显著;同时TGF-β1显著增加,且Vimentin的表达增加,E-cadherin的表达明显降低,Western blot检测Vimentin、E-cadherin结果与免疫组化一致,且α-SMA也显著升高。结论高腺嘌呤饮食可以建立CKD并发RIF小鼠模型。Sigirr的缺失增加肾间质IL-1β介导NF-κB信号通路的活化,促进TGF-β1表达,有利于上皮-间质细胞转分化相关RIF过程。Objective To investigate the role and mechanism of Sigirr deletion in chronic kidney disease complicated with renal interstitial fibrosis(CKD-RIF)in mice.Methods polymerase chain reaction(PCR)was used for identification of gene types of mice.Mice were continuously fed with the foods containing 0.2%adenine for 12 weeks to establish the CKD-RIF models.Then,serum was collected to detect levels of creatinine and nitrogen when mice were killed.H&E staining was used to analyze the pathological changes of kidney tissues.Masson staining was used to observe the degree of renal fibrosis.Immunohistochemistry was used to detect the changes of the interest proteins,such as IL-1β,MyD88,activated NF-κB,TGF-β1,E-cadherin and Vimentin.Results Serum creatinines and urea nitrogens of mice fed with high adenine(CKD-RIF groups)significantly increased,compared with those of the control groups.H&E and Masson staining results showed that there were more infiltrated inflammatory cells and more critical collagen fiber deposition in the renal tissues of the Sigirr^(-/-)mice with CKD-RIF.Western blot and Immunohistochemical analysis showed that the expression of IL-1βand its downstream MyD88 increased,and the level of phosphorylated NF-κB(p-P65)significantly increased in the renal tissues of CKD-RIF mice compared with the controls.And upregulation of these proteins in renal tissues of Sigirr^(-/-)mice with CKD-RIF was more obvious than that of the CKD-RIF Sigirr^(+/+)mice.TGF-β1,as a key cytokine involved in renal interstitial fibrosis,significantly increased,followed by the increase of vimentin,as well as the decrease of E-cadherin.The results of vimentin and cadherin E detected by Western blot were consistent with those of immunohistochemistry,andα-SMA also increased significantly.Conclusion Adenine diet successfully induces CKD-RIF mice models.Sigirr deletion is beneficial to activation of the IL-1βmediating NF-κB signal pathway,which promotes TGF-β1 expression in the renal interstitiums to induce renal interstitial fibrosis.

关 键 词：Sigirr基因 慢性肾病 肾间质纤维化 IL-1Β NF-ΚB 

分 类 号：R34[医药卫生—基础医学]