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发布于:2020-11-6 17:01:33  访问:2024 次 回复:0 篇
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Myeloperoxidase , malondialdehyde , superoxide dismutase , catalase , decreased glutathione and organo gold (from Youtube) glutathione peroxidase (GSH-Px) had been detected for evaluating the effect of GLPP on IR-mediated oxidative anxiety in the kidney. Compared with the sham group, RIRI considerably increased the levels of MPO and MDA and decreased the activities of SOD, CAT, GSH and GSH-Px while GLPP reversed these modifications caused by IR (Fig. 2A–F).
The H/R considerably decreased the activities of SOD and GSH, which was associated with a reciprocal raise in MDA level and ROS production (Fig. 5C,D). A CCK-8 assay was utilised to assess the cytotoxicity of GLPP. At concentrations from 1.1 μg/ml to 810 μg/ml, GLPP showed no clear cytotoxicity on NRK-52E cells (Fig. 5A). Hypoxia for 12 h followed by reoxygenation for 1 h drastically decreased the cell viability compared to manage cells. Pretreatment with GLPP improved cell viability in a dose-dependent manner (Fig. 5B).
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Impaired redox status final results in accumulation of ROS, thus activates mitochondrial and ER stress. When homeostasis is disrupted and adaptive responses fail to compensate for the tension, apoptosis is triggered. A TUNEL assay was utilized to evaluate apoptosis in renal tissues induced by IR. A lot more apoptotic cells appeared in kidneys subjected to IR than in the sham-operated kidneys. GLPP reduced IR-induced TUNEL-optimistic cells by 21.75%, which suggests that GLPP protects kidneys from renal tubular apoptosis (Fig. 3A).
Final results had been confirmed by Western blot analysis, demonstrated as decreased ratios of p-p53/p53 and cleaved caspase-3/caspase-3 in the IR GLPP-treated group (Fig. 3B). Oxidative anxiety plays a crucial part in the activation of mitochondrial and ER anxiety, which benefits in a series of abnormalities such as apoptosis, necrosis and other significant consequences.
We then detected the expression of manganese superoxide dismutase (Mn-SOD), which is an critical cellular antioxidant enzyme. 2G, IR significantly decreased the expression of Mn-SOD while GLPP improved its expression. Also, we isolated cell membrane and cytosol proteins separately for evaluating the adjustments of p47phox, a core regulatory subunit of NADPH oxidase, to market NADPH oxidase-dependent production of ROS. We additional explored irrespective of whether postoperative administration of GLPP protected against RIRI. GLPP was intraperitoneally administered at the beginning of reperfusion.
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In line with the mouse model, we located that H/R elevated cell apoptosis by signifies of up-regulating of the ratios of p-p53/p53 and cleaving capsase-3/caspase-three. However, pretreatment with GLPP reversed these ratios in a dose-dependent manner, which confirmed the outcomes of the TUNEL evaluation.
All these information recommend that GLPP has a protective impact against H/R-induced apoptosis in renal tubular cells. We then studied the effect of GLPP against cellular oxidative strain.
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