Kim Yosep, Kim Jongwan, Han Sang Jun
Department of Biotechnology, College of Fisheries Sciences, Pukyong National University, Busan, Republic of Korea.
Department of Medical Laboratory Science, Dong-Eui Institute of Technology, Busan, Republic of Korea.
Kidney Res Clin Pract. 2023 Mar;42(2):188-201. doi: 10.23876/j.krcp.22.033. Epub 2023 Mar 22.
Diminazene aceturate (DIZE), an angiotensin-converting enzyme 2 (ACE2) activator, exerts anti-inflammatory and antifibrotic effects in a variety of human chronic diseases. However, the role of DIZE in kidney fibrosis and the underlying mechanism remain unclear. Therefore, we investigated the effects of DIZE on the progression of renal fibrosis after unilateral ureteral obstruction (UUO), a well-established model of chronic kidney disease.
C57BL/6 female or male mice were subjected to right UUO. Mice received 15 mg/kg DIZE or vehicle (saline) daily. On the 7th day after UUO, kidneys were collected for analysis of renal fibrosis (α-smooth muscle actin, phosphorylated SMAD3, transforming growth factor (TGF)-β, Masson's trichrome, and Sirius red staining), inflammation (macrophage infiltration, proinflammatory cytokines/ chemokines), apoptosis/necrotic cell death (TUNEL and periodic acid-Schiff staining), and ACE2 activity and messenger RNA (mRNA) expression.
Treatment with DIZE exacerbated renal fibrosis by upregulating the profibrotic TGF-β/SMAD3 pathway, proinflammatory cytokine/chemokines (interleukin [IL]-1β, monocyte chemoattractant protein-1, IL-6, and macrophage inflammatory protein-2) levels, M2 macrophage accumulation (CD206, IL-4, IL-10, and CX3CL1), and apoptotic/necrotic cell death in the obstructed kidneys of female mice but not male mice. However, DIZE treatment had no effect on ACE2 activity or mRNA expression.
DIZE exacerbates UUO-induced renal fibrosis by aggravating tubular damage, apoptosis, and inflammation through independent of angiotensin (1-7), angiotensin II levels, and ACE2 expression/activity, rather than protecting against renal fibrosis after UUO. DIZE also has powerful effects on recruiting macrophages, including the M2-polarized subtype, in female UUO mice.
乙酰氧肟酸二脒那嗪（DIZE）是一种血管紧张素转换酶2（ACE2）激活剂，在多种人类慢性疾病中发挥抗炎和抗纤维化作用。然而，DIZE在肾纤维化中的作用及其潜在机制仍不清楚。因此，我们研究了DIZE对单侧输尿管梗阻（UUO）后肾纤维化进展的影响，UUO是一种成熟的慢性肾脏病模型。
对C57BL/6雌性或雄性小鼠进行右侧UUO手术。小鼠每天接受15mg/kg DIZE或载体（生理盐水）。UUO术后第7天，收集肾脏用于分析肾纤维化（α-平滑肌肌动蛋白、磷酸化SMAD3、转化生长因子（TGF）-β、Masson三色染色和天狼星红染色）、炎症（巨噬细胞浸润、促炎细胞因子/趋化因子）、凋亡/坏死性细胞死亡（TUNEL和过碘酸希夫染色）以及ACE2活性和信使核糖核酸（mRNA）表达。
DIZE治疗通过上调促纤维化的TGF-β/SMAD3途径、促炎细胞因子/趋化因子（白细胞介素[IL]-1β、单核细胞趋化蛋白-1、IL-6和巨噬细胞炎性蛋白-2）水平、M2巨噬细胞积聚（CD206、IL-4、IL-10和CX3CL1）以及雌性小鼠而非雄性小鼠梗阻肾脏中的凋亡/坏死性细胞死亡，加剧了肾纤维化。然而，DIZE治疗对ACE2活性或mRNA表达没有影响。
DIZE通过加重肾小管损伤、凋亡和炎症，独立于血管紧张素（1-7）、血管紧张素II水平和ACE2表达/活性，加剧UUO诱导的肾纤维化，而不是预防UUO后的肾纤维化。DIZE对雌性UUO小鼠招募巨噬细胞（包括M2极化亚型）也有强大作用。
