尿酸单钠(MSU)结晶沉积在局部组织和器官诱导炎症反应,导致痛风等疾病。MSU已被认为是各种临床病症中常见且普遍的病理。在这项研究中,我们研究了MSU在糖尿病肾病(DKD)发病机制中的作用.我们使用链脲佐菌素(STZ)诱导糖尿病肾病小鼠的肾损伤,并使用Masson三色染色和胶原蛋白III免疫荧光染色评估肾组织病理学损伤。我们测量了丙二醛(MDA)的水平,超氧化物歧化酶(SOD),和尿酸(UA)使用ELISA。NLRP3、p-NF-κB蛋白表达水平,通过Western印迹分析SHP2、p-STAT3和p-ERK1/2。进一步探讨MSU在糖尿病肾病中的作用,我们进行了体外实验。在我们的体内实验中,我们发现与模型组相比,用MSU治疗后,小鼠肾脏间质纤维化显着增加,伴随着MDA水平的升高,SOD,UA。此外,NLRP3,p-NF-NB的蛋白表达,SHP2、p-STAT3和p-ERK1/2上调。在我们随后对小鼠成纤维细胞(L929细胞)的研究中,我们发现高葡萄糖,MSU,TGF-β能促进P22、GP91、NLRP3、NF-κB的表达,p-NF-κB,p-SHP2,p-EGFR,p-STAT3和胶原-III蛋白。此外,我们发现SHP2可以抵消MSU对p-SHP2、p-EGFR表达的上调趋势,p-STAT3和胶原蛋白III蛋白,和抑制剂YQ128,NAC,和西妥昔单抗表现出相似的效果。此外,免疫荧光结果显示SHP2可抑制L929细胞纤维化标志物α-SMA的表达。这些发现表明,MSU可以促进肾成纤维细胞SHP2的表达,诱导氧化应激,激活NLRP3/NF-κB通路,通过TGFβ/STAT3/ERK1/2信号通路促进糖尿病肾病成纤维细胞增殖,导致肾脏纤维化。
Monosodium urate (MSU) crystallisation deposited in local tissues and organs induce inflammatory reactions, resulting in diseases such as gout. MSU has been recognized as a common and prevalent pathology in various clinical conditions. In this study, we investigated the role of MSU in the pathogenesis of diabetic kidney disease (DKD). We induced renal injury in diabetic kidney disease mice using streptozotocin (STZ) and assessed renal histopathological damage using Masson\'s trichrome staining and Collagen III immunofluorescence staining. We measured the levels of malondialdehyde (MDA), superoxide dismutase (SOD), and uric acid (UA) using ELISA. Protein expression levels of NLRP3, p-NF-κB, SHP2, p-STAT3, and p-ERK1/2 were analyzed by Western blot. To further investigate the role of MSU in diabetic kidney disease, we conducted in vitro experiments. In our in vivo experiments, we found that compared to the Model group, there was a significant increase in interstitial fibrosis in the kidneys of mice after treatment with MSU, accompanied by elevated levels of MDA, SOD, and UA. Furthermore, the protein expression of NLRP3, p-NF-NB, SHP2, p-STAT3, and p-ERK1/2 was upregulated. In our subsequent studies on mouse fibroblasts (L929 cells), we discovered that high glucose, MSU, and TGF-β could promote the expression of P22, GP91, NLRP3, NF-κB, p-NF-κB, p-SHP2, p-EGFR, p-STAT3, and Collagen-III proteins. Additionally, we found that SHP2 could counteract the upregulation trend induced by MSU on the expression of p-SHP2, p-EGFR, p-STAT3, and Collagen-III proteins, and inhibitors YQ128, NAC, and Cetuximab exhibited similar effects. Furthermore, immunofluorescence results indicated that SHP2 could inhibit the expression of the fibrosis marker α-SMA in L929 cells. These findings suggest that MSU can promote renal fibroblast SHP2 expression, induce oxidative stress, activate the NLRP3/NF-κB pathway, and enhance diabetic kidney disease fibroblast proliferation through the TGFβ/STAT3/ERK1/2 signaling pathway, leading to renal fibrosis.