The superconducting magnet with a large-gradient, high magnetic field (LG-HMF) and its specifications.

<p>The superconducting magnet can generate a magnetic force field of −1360, 0, and 1312 T²/m in a 51 mm (diameter) room-temperature bore, corresponding to 3 apparent body force levels (μ-g, 1-g, and 2-g) and 3 magnetic induction intensities (12 T, 16 T, and 12 T), respectively. The height of the superconducting magnet is 195 cm with a 51 mm × 450 mm (diameter) cylindrical cavity.</p

Detection of the effects of LG-HMF on MC3T3-E1 (A–L) and MG-63 (M–X) morphology by transmission electron microscopy (TEM).

<p>After MG-63 and MC3T3-E1 cells were cultured in the bore of LG-HMF (μ-g, 1-g, and 2-g) at 37°C for 24 h, the osteoblasts were fixed with 2.5% glutaraldehyde, then dehydrated by gradient acetone and embedded. Ultra-thin sections were contrasted according to Reynolds and examined in a JEOL analytical transmission electron microscope. MC: mitochondria, ER: endoplasmic reticulum, GP: glycogen particles, NMC: nucleus membrane cavity, CSK: cytoskeleton, HC: heterochromatin, MV: microvilli, RER: rough endoplasmic reticulum, ECM: extracellular matrix, AV: autophagic vacuoles. N: A–L: MC3T3-E1cells, M–X: MG-63 cells. A–C and M–O: control, D–F and P–R: μ-g, G–I and S–U: 1-g, J–L and V–X: 2-g.</p

In cell western (ICW) analysis of the effects of LG-HMF on collagen I and fibronectin (FN) expression in MC3T3-E1 (A, B) and MG-63 (C, D).

<p>After 24 h of exposure to LG-HMF, the expression of collagen I and FN MG-63 and MC3T3-E1 cells were examined by ICW. The results indicated that collagen I expression significantly increased in MC3T3-E1 cells under μ-g compared to 2-g conditions and 1-g compared to control conditions (<i>P</i><0.05), and in MG-63 cells collagen I expression increased under 1-g compared to control conditions (<i>P</i><0.05). FN expression was also up-regulated in MG-63 cells under 1-g compared to control conditions (<i>P</i><0.05). *: <i>P</i><0.05, **: <i>P</i><0.01.</p

MTT analysis of the effects of LG-HMF on the viability of MC3T3-E1 (A–C) and MG-63 (D–F) cells.

<p>After MG-63 and MC3T3-E1 cells were cultured at 37°C for 12 h, 24 h, and 48 h in the bore of the superconducting magnet, the cells were removed from LG-HMF. The MTT assay was used to detect the effects of LG-HMF (μ-g, 1-g, and 2-g) on cell viability. Cell viabilities increased slightly in both cell lines compared to control conditions after 24 h or 48 h of exposure to LG-HMF (<i>P</i><0.05). A–C: MG-63 cells; D–F: MC3T3-E1cells. A, D: exposure to LG-HMF for 12 h; B, E: exposure to LG-HMF for 24 h; C, F: exposure to LG-HMF for 48 h. *: <i>P</i><0.05, **: <i>P</i><0.01, ***: <i>P</i><0.001.</p

PNPP analysis of the effects of LG-HMF on MC3T3-E1 (A) and MG-63 (B) ALP production.

<p>A colorimetric p-nitrophenyl phosphate assay was used to measure ALP expression in osteoblasts after 24 h of exposure to the LG-HMF environment. p-NP was quantified based on the spectrophotometric absorbance at 405 nm. The results indicate that the effects of the magnetic field on ALP were dominant in MC3T3-E1 cells or MG-63 cells. *: <i>P</i><0.05, **: <i>P</i><0.01.</p

Effect of ACS84 on oxidative stress induced by 6-OHDA in SH-SY5Y cells.

<p>(A) Dose dependent effect of ACS84 on ROS generation in the 6-OHDA-treated (50 µM) SH-SY5Y cells. Cells were pretreated with ACS84 at different concentrations for 4 h. DCFDAH₂ (10 µM) was given 30 min before the addition of 6-OHDA (50 µM). The results were obtained after the treatment with 6-OHDA for 1 h. (B–C) Effect of ACS84, L-Dopa and NaHS at 10 µM on ROS generation (B) and SOD (C) in SH-SY5Y cells treated with 6-OHDA. SOD activity was measured 4 h after 6-OHDA treatment. Data are presented as mean ± SEM, n = 4–8, ^###<i>P</i><0.001 versus control; *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001, versus 6-OHDA-treated cells; ^†††<i>P</i><0.001, versus ACS84-treated cells.</p

Effect of ACS84 on oxidative stress in the striatum of unilateral 6-OHDA-lesioned PD rat model.

<p>ACS84 treatment (10 mg kg⁻¹ day⁻¹, i.g) alleviated the increased MDA production. Data are presented as mean ± SEM, n = 4–6. *<i>P</i><0.05 versus lesion site of Sham group and ^#<i>P</i><0.05 versus lesion site of Vehicle group.</p

Effect of ACS84 on dopamine and its metabolites in 6-OHDA-lesioned striatum.

<p>The concentration of dopamine and its metabolites in 6-OHDA-lesioned striatum were measured using HPLC. Units for Dopamine, DOPAC and HVA concentrations were ng/g tissue. Data are presented as mean ± SEM, n = 6–8.</p>#<p>p<0.05 versus Sham group and *p<0.05 versus Vehicle group.</p

Effect of ACS84 on 6-OHDA-induced TH+ neuronal degeneration.

<p>Immunofluorescence staining showed that ACS84 (10 mg kg⁻¹ day⁻¹, i.g) alleviated TH+ neuron loss in SN of 6-OHDA-lesioned PD rats. Photos were taken at x100 magnification, and the white bar indicated 0.1 µm. Samples were collected from two independent experiments.</p

Nrf2 Signaling Contributes to the Neuroprotective Effects of Urate against 6-OHDA Toxicity

<div><p>Background</p><p>Mounting evidence shows that urate may become a biomarker of Parkinson's disease (PD) diagnosis and prognosis and a neuroprotectant candidate for PD therapy. However, the cellular and molecular mechanisms underlying its neuroprotective actions remain poorly understood.</p><p>Results</p><p>In this study, we showed that urate pretreatment protected dopaminergic cell line (SH-SY5Y and MES23.5) against 6-hydroxydopamine (6-OHDA)- and hydrogen peroxide- induced cell damage. Urate was found to be accumulated into SH-SY5Y cells after 30 min treatment. Moreover, urate induced NF-E2-related factor 2 (Nrf2) accumulation by inhibiting its ubiquitinationa and degradation, and also promoted its nuclear translocation; however, it did not modulate Nrf2 mRNA level or Kelch-like ECH-associated protein 1 (Keap1) expression. In addition, urate markedly up-regulated the transcription and protein expression of γ-glutamate-cysteine ligase catalytic subunit (γ-GCLC) and heme oxygenase-1 (HO-1), both of which are controlled by Nrf2 activity. Furthermore, Nrf2 knockdown by siRNA abolished the intracellular glutathione augmentation and the protection exerted by urate pretreatment.</p><p>Conclusion</p><p>Our findings demonstrated that urate treatment may result in Nrf2-targeted anti-oxidant genes transcription and expression by reducing Nrf2 ubiquitination and degradation and promoting its nuclear translocation, and thus offer neuroprotection on dopaminergic cells against oxidative stresses.</p></div