Vukovic Andrija, Karanovic Danijela, Mihailovic-Stanojevic Nevena D, Miloradovic Zoran, Brkic Predrag, Zivotic Maja, Nesovic Ostojic Jelena, Ivanov Milan, Kovacevic Sanjin, Vajic Una-Jovana, Jovovic Djurdjica, De Luka Silvio R
Institute of Pathological Physiology, Faculty of Medicine, University of Belgrade, Dr Subotića 1, 11000 Belgrade, Serbia.
Institute for Medical Research, National Institute of Republic of Serbia, University of Belgrade, Dr Subotića 4, 11000 Belgrade, Serbia.
Biomedicines. 2024 Dec 9;12(12):2788. doi: 10.3390/biomedicines12122788.
BACKGROUND/OBJECTIVES: Chronic kidney disease (CKD) is a progressive pathological condition which results in the severe fibrosis of the kidneys. However, the mechanisms of CKD progression and fibrogenesis remain unclear. We wanted to examine the effects that apocynin and hyperbaric oxygen therapy (HBOT) have on renal function and structure in animals with CKD induced through 5/6 nephrectomy (5/6 Nx-L).
Male Wistar rats were divided in 5 groups (n = 8/group) as follows: control-sham-operated rats; Nx-L-rats with 5/6 Nx-L; APO-5/6 Nx-L + apocynin treatment; HBOT-5/6 Nx-L + hyperbaric oxygen treatment, and APO+HBOT-5/6 Nx-L, treated with both treatments. All treatments started 4 weeks after the final step of CKD induction and lasted for 4 weeks. At the end of the experiment, urine samples were collected for the proteinuria assessment and the mean arterial pressure (MAP) was measured. Kidneys were collected for histopathological, Western blot, and immunohistochemical analyses.
All treatments significantly decreased MAP compared to the Nx-L group ( < 0.001). In the APO and APO+HBOT groups, the level of proteinuria was decreased compared to the Nx-L group ( < 0.05 and < 0.01, respectively). All examined treatments significantly decreased the intensity of lesions in the kidney compared to those observed in the Nx-L group ( < 0.001). Isolated treatments with apocynin and HBOT induced a significant decrease in desmin expression compared to the Nx-L group ( < 0.05); meanwhile, they did not affect the levels of fibronectin (FN) and hypoxia-inducible factor-1α (HIF-1α). Combined treatment did not affect desmin expression levels; however, it induced a significant increase in fibronectin expression compared to Nx-L ( < 0.001).
Apocynin treatment decreased BP and protein loss, and it improved renal morphology at least partly through the downregulation of desmin expression without changing FN and HIF-1α. Hyperbaric oxygen therapy improved hypertension but failed to significantly affect the level of proteinuria. Combined treatment (apocynin and HBOT) normalized blood pressure (BP) values, renal function, and improved kidney structure by modulating FN and HIF-1α, without affecting desmin protein expression. Further studies are needed to elucidate the mechanisms of slowing down the progression of CKD in this experimental model.
背景/目的：慢性肾脏病（CKD）是一种进行性病理状态，可导致肾脏严重纤维化。然而，CKD进展和纤维化形成的机制仍不清楚。我们想研究阿朴吗啡和高压氧疗法（HBOT）对通过5/6肾切除术（5/6 Nx-L）诱导的CKD动物的肾功能和结构的影响。
将雄性Wistar大鼠分为5组（每组n = 8），如下：假手术对照组大鼠；5/6 Nx-L诱导的CKD大鼠；APO-5/6 Nx-L + 阿朴吗啡治疗组；HBOT-5/6 Nx-L + 高压氧治疗组，以及APO+HBOT-5/6 Nx-L组，接受两种治疗。所有治疗在CKD诱导最后一步后4周开始，持续4周。实验结束时，收集尿液样本进行蛋白尿评估，并测量平均动脉压（MAP）。收集肾脏进行组织病理学、蛋白质印迹和免疫组织化学分析。
与Nx-L组相比，所有治疗均显著降低了MAP（< 0.001）。在APO和APO+HBOT组中，与Nx-L组相比，蛋白尿水平降低（分别为 < = 0.05和 < = 0.01）。与Nx-L组相比，所有检查的治疗均显著降低了肾脏病变的严重程度（< 0.001）。与Nx-L组相比，单独使用阿朴吗啡和HBOT治疗可显著降低结蛋白表达（ < 0.05）；同时，它们不影响纤连蛋白（FN）和缺氧诱导因子-1α（HIF-1α）的水平。联合治疗不影响结蛋白表达水平；然而，与Nx-L组相比，它诱导纤连蛋白表达显著增加（< 0.001）。
阿朴吗啡治疗可降低血压和蛋白质丢失，并至少部分通过下调结蛋白表达改善肾脏形态，而不改变FN和HIF-1α。高压氧疗法可改善高血压，但未能显著影响蛋白尿水平。联合治疗（阿朴吗啡和HBOT）通过调节FN和HIF-1α使血压（BP）值、肾功能正常化，并改善肾脏结构，而不影响结蛋白的表达。需要进一步研究以阐明在该实验模型中减缓CKD进展的机制。
