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Objective: To investigate the role of Nox2 on AngII-stimulated aldosterone biosynthesis. Methods: H295R cells were cultured and randomly divided into control group, Angll group, AngII+gp91ds-tat group, AngII+PEG-Cat group, AngII+Nox2 siRNA group. Q-PCR and western blot were performed to determine CYP1IB2 and Nox2 expression in H295R cells. Aldosterone level in cells culture supernatant was determined by radioimmunoassay. Nox2-derived ROS and H2O2 production in H295R cells were determined by flow cytometric analysis and fluorescence microplate reader. Results: 10 nmol/L Angll increased cellular ROS and H2O2 generation, increased the Nox2 and CYP11B2 expression and accelerated aldosterone biosynthesis in a time-dependent manner (P<0.05). And AngII-induced cellular ROS and H₂O, production in both AngII+gp91ds-tat and AngII+PEG-Cat groups were lower than those in AngII group (P<0.05). However, there were no difference in inhibition of cellular ROS and H2O2 between treatment with gp91ds-tat and PEG-Cat (P>0.05). The expression of CYP11B2 in H295R cells was up-regulated after treated with 10 nmol/L Angll for 24 h (P<0.05), while gp91ds-tat, PEG-Cat and Nox2 siRNA significantly abrogated the CYP11B2 expression in H295R cells (P<0.05). Conclusion: Nox2 dependent ROS play an important role in aldosterone synthesis. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of liraglutide (Lir) on microglia/macrophage polarization and on the expression of nicotinamide adenine dinucleotide phosphate oxidase (NOX2) in the ischemic brain tissue of diabetic rats with focal cerebral ischemia. Method Seventy-five rats were randomly and equally divided into 5 groups: cerebral ischemia (CI) group, diabetes+cerebral ischemia (DCI) group, insulin treatment (Ins) group, Lir treatment (Lir) group, and NOX2 agonist tetrabromocinnamic acid (TBCA) + Lir group (TBCA). On the 3rd day after cerebral ischemia, the neurological deficit scores of the rats were evaluated, and 2, 3, 5-triphenyltetrazolium chloride (TTC) staining was adopted to measure the cerebral infarct volume in each group. The co-labeling of CD16/32 and CD206 with Iba1 on microglia/macrophages were detected by immunofluorescence double-label staining, and the protein expression of NOX2, TNF-α, IL-1β, IL-10 and TGF-β were determined using Western blotting. Results As compared with the DCI group, the neurological deficit scores were decreased, and the cerebral infarction volume was reduced in the Lir group. The cell number of Iba1+/CD16/32+ microglia/macrophage (M1 type) was declined, with down-regulated expression of TNF-α, IL-1β and NOX2 in the Lir group, while the number of Iba1+/CD206+ microglia /macrophage (M2 type) was increased, with improved levels of IL-10 and TGF-β (all P < 0.05). However, the above effects were reversed by the treatment of NOX2 agonist TBCA (P < 0.05). Conclusion The neuroprotective effects of Lir on cerebral ischemic injury may be related to the inhibition of NOX2 expression and promotion of microglia/macrophages conversion to M2 type.

Objective: To determine whether NADPH oxidase II(NOX2) is associated in berberine(BBR)-induced protection on HT22 neuronal cells exposed to β amyloid(Aβ). Methods: HT22 neuronal cells were divided into five groups, including the normal cultured Control group, 10 μM Aβ exposure group(Aβ), 1 μM BBR plus 10 μM Aβ treatment group(BBR+Aβ), NOX2-siRNA treatment group(NOX2-siRNA+BBR+Aβ) and scrambled(SC)-si RNA group(SC-si RNA+BBR+Aβ), after an incubation for 24 h, methylthiazolyldiphenyl-tetrazolium(MTT) was used to assess cell viability, lactic dehydrogenase(LDH) reagent kit was used to evaluate the LDH release in the medium, Western blot was taken to evaluate the apoptosis-associated protein Cleaved caspase-3 and NOX2 expressions, reactive oxygen species(ROS) and malondialdehyde(MDA) reagent kits were used to assess the intracellular ROS and MDA levels respectively. Results: Compared with the control group, 10 μM Aβ reduced the viability of HT22 cells, increased LDH activity, intracellular ROS and MDA levels, upregulated cleaved caspase-3 and NOX2 expressions(P<0.05), and 1 μM BBR treatment significantly inhibited the Aβ-induced injury and oxidative effects mentioned above(P<0.05). However, NOX2-siRNA treatment obviously abrogated BBR-induced neuroprotection and anti-oxidative effects(P<0.05), and SC-si RNA showed no significant effect on BBR-mediated neuroprotective effects(P>0.05). Conclusion: NOX2 may mediate BBR-induced neuroprotective effects on the HT22 neuronal cells exposed to Aβ. [ABSTRACT FROM AUTHOR]

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Objective: To explore the difference of therapeutic effect between tert-butylhydroquinone (tBHQ) and sulforaphane (SFN) in rats with traumatic brain injury. Methods: A total of SO healthy adult male rats were randomly divided into 4 groups: sham group. TBI group, tBHQ treatment group and SFN treatment group, then the traumatic brain injury (TBI) model were prepared by eCCI. tBHQ treatment group rats were given intraperitoneal injection three tunes tBHQ (50 mg/kg) before injury 24 h. and SFN treatment group rats were given intraperitoneally injected SFN (5 mg/kg) after injury 15 min. Neurologic deficits was evaluated 24 h after TBI by mNSS method, brain edema was detected by the wet-dry ratio method. The expression levels of NOX2 protein and Nrf2 protein in the brain tissue were investigated using Western blot and ELISA analysised. respectively. Results: Compared with TBI group, there were significant difference on mNSS scores ((4.5± 0.71) vs (9.2± 0.79). (6.0± 0.82) vs (9.2± 0.79)). brain edema (79.4% vs 85.6%. 80.3% vs 85.6%). the expression levels of NOX2 protein and Nrf2 protein (0.93 ng/mL vs 0.81 ng/mL. 0.87 ng/mL vs 0.81 ng/mL) in tBHQ treatment group and SFN treatment group. In addition, there were also significant differences on mNSS scores ((4.5± 0.71) vs (6.0± 0.82)), the expression levels of NOX2 protein and Nrf2 protein (0.93 ng/mL vs 0.87 ng/mL) between tBHQ treatment group and SFN treatment group. Conclusion: In the TBI model of rats. tBHQ and SFN could effectively attenuated cerebral oxidative stress and improved nerve function after TBI. but the curative effect of tBHQ was better than the SFN. [ABSTRACT FROM AUTHOR]

摘 要:【目的】 观察miR-210 inhibitor对大鼠背脊髓神经元细胞氧化应激的保护作用?【方法】 体外培养大鼠背脊髓神经元细胞,随机分成正常组,H2O2组,H2O2联合miRNA Negative control组以及H2O2联合miR-210 inhibitor组?采用MTT法检测细胞的存活率,流式细胞仪检测细胞的凋亡率,活性氧检测试剂盒检测细胞内活性氧,Elisa检测NOX2?【结果】 qPCR结果显示,100 μmol/L H2O2处理可导致大鼠背脊髓神经元细胞内miR-210表达量上升7.34倍;转染miR-210 inhibitor可明显抑制miR-210的表达量;MTT结果显示,抑制miR-210的表达可明显降低H2O2对大鼠背脊髓神经元细胞存活率的抑制作用;流式细胞仪检测发现抑制miR-210的表达可以明显阻止H2O2对大鼠背脊髓神经元细胞凋亡的诱导,显著降低凋亡率;进一步研究发现,转染miR-210 inhibitor可显著性降低H2O2诱导的细胞内NOX2和ROS的表达?【结论】 H2O2刺激鼠脊髓神经元细胞导致细胞内miR-210表达的上调;降低miR-210的表达能够降低细胞内NOX2和ROS的表达,抑制NOX2/ROS通路从而减缓H2O2对大鼠背脊髓神经元细胞造成的损伤,推测miR-210可以作为脊髓损伤治疗的靶点?