Parkinson's disease (PD) is a common neurodegenerative disorder associated with striatal dopaminergic neuronal loss in the Substantia nigra. Oxidative stress plays a significant role in several neurodegenerative diseases. Paraquat (PQ) is considered a potential neurotoxin that affects the brain leading to the death of dopaminergic neurons mimicking the PD phenotype. Various scientific reports have proven that cryptotanshinone possesses antioxidant and anti-inflammatory properties. We hypothesized that cryptotanshinone could extend its neuroprotective activity by exerting antioxidant effects. This study was designed to evaluate the effects of cryptotanshinone in both cellular and animal models of PQ-induced PD. Annexin V-PI double staining and immunoblotting were used to detect apoptosis and oxidative stress proteins, respectively. Reactive oxygen species kits were used to evaluate oxidative stress in cells. For in vivo studies, 18 B6 mice were divided into three groups.The rotarod data revealed the motor function and immunostaining showed the Yu-Jung Lin and Chih-Yang Huang contributed equally to this work.

Traumatic optic neuropathy (TON) may cause severe visual loss following direct or indirect head trauma which may result in optic nerve injuries and therefore contribute to the subsequent loss of retinal ganglion cells by inflammatory mediators and reactive oxygen species (ROS). Granulocyte colony-stimulating factor (G-CSF) provides the anti-inflammatory and anti-oxidative actions but has a short half-life and also induces leukocytosis upon typical systemic administration. The purpose of the present study was to investigate the relationship between the anti-oxidative response and neuroprotective effects of long-acting pegylated human G-CSF (PEG-G-CSF) in a rat model of optic nerve crush (ONC). Adult male Wistar rats (150–180 g) were chosen to have a sham operation in one eye and have ONC in the other. PEG-G-CSF or phosphate-buffered saline (PBS control) was immediately administered after ONC by intravitreal injection (IVI). We found the IVI of PEG-G-CSF does not induce systemic leukocytosis, but increases survival of RGCs and preserves the visual function after ONC. TUNEL assays showed fewer apoptotic cells in the retina in the PEG-G-CSF-treated eyes. The number of sorely ED1-positive cells was attenuated at the lesion site in the PEG-G-CSF-treated eyes. Immunoblotting showed up-regulation of p-Akt1, Nrf2, Sirt3, and HO-1 in the ON of the PEG-G-CSF-treated eyes. Our results demonstrated that one IVI of long-acting PEG-G-CSF is neuroprotective in the rONC. PEG-G-CSF activates the p-Akt1/Nrf2/Sirt3 and the p-Akt1/Nrf2/HO-1 axes to provide the antioxidative action and further attenuated RGC apoptosis and neuroinflammation. This provides crucial preclinical information for the development of alternative therapy with IVI of PEG-G-CSF in TON.

Supplementing with vitamin B3 has been reported to protect against retinal ganglion cell (RGC) damage events and exhibit multiple neuroprotective properties in a mouse model of optic nerve injury. In this study, a rat model of anterior ischemic optic neuropathy was used to assess the neuroprotective benefits of vitamin B3 (rAION). Vitamin B3 (500 mg/kg/day) or phosphate-buffered saline (PBS) was administered to the rAION-induced rats every day for 28 days. The vitamin B3-treated group had significantly higher first positive and second negative peak (P1-N2) amplitudes of flash visual-evoked potentials and RGC densities than the PBS-treated group (p < 0.05). A terminal deoxynucleotidyl transferase dUTP nick end labeling assay conducted on vitamin B3-treated rats revealed a significant reduction in apoptotic cells (p < 0.05). Superoxide dismutase and thiobarbituric acid reactive substance activity showed that vitamin B3 treatment decreased reactive oxygen species (p < 0.05). Therefore, vitamin B3 supplementation preserves vision in rAION-induced rats by reducing oxidative stress, neuroinflammation, and mitochondrial apoptosis.

An ischemic insult at optic nerve (ON) is followed by detrimental neuroinflammation that results in progressive and long‐lasting retinal ganglion cell (RGC) death and vision loss. Icariin was reported to be a safe and effective natural anti‐inflammatory drug. Herein, we evaluated the long‐term therapeutic effects of a single intravitreal injection of poly(lactide‐co‐glycolide) PLGA–icariin in a rat model of anterior ischemic optic neuropathy (rAION). Treatment with PLGA microspheres of icariin preserved the visual function and RGC density for 1 month in the rAION model. In addition, ON edema and macrophage infiltration were inhibited by treating PLGA microspheres of icariin. We found that the binding complex of icariin and CCAAT enhancer binding protein beta (CEBP‐β) significantly induced endogenous granulocyte colony‐stimulating factor (G‐CSF) expression to activate noncanonical nuclear factor kappa B (NF‐κB) signaling pathway by promoting an alternative phosphorylation reaction of IKK‐β. Activation of noncanonical NF‐κB signaling pathway promoted the M2 microglia/macrophage polarization and AKT1 activation, which prevented neuroinflammation and RGC apoptosis after ON infarct. This study concluded that protective mechanism of icariin is a CEBP‐β/G‐CSF axis‐induced noncanonical NF‐κB activation, which provides the long‐term neuroprotective effects via anti‐inflammatory and antiapoptotic actions after ON ischemia.

BackgroundAn ischemic insult at the optic nerve is followed by detrimental neuroinflammation that results in the loss of the retinal ganglion cells (RGCs) and vision. NF-κB is the key transcription factor of inflammatory cytokines in response to neuroinflammatory events. Icariin, an anti-inflammatory drug, is involved in the regulation of NF-κB activation. However, the protective mechanisms of icariin-induced NF-κB activation remain little known. Here, the protective mechanisms of icariin-induced NF-κB activation were investigated in experimental optic nerve ischemia. MethodsThe GOLD bio-informatics tool was used to select target drug based on CEBP-β binding affinity. The neuroprotective effects of target drug, Icariin, were assessed by measuring the flash visual electrode potentials, density of fluorogold-labeled cells, and TUNEL-positive cells in ganglion cell layer. The processes affecting RGC death were probed using optical coherence tomography, immunohistochemistry and immunoblotting techniques. ResultsThe simulation analysis and in vivo test demonstrated that the binding complex of icariin and CEBP-β significantly induced endogenous G-CSF expression. A single intravitreal injection of poly(lactic-co-glycolic acid) (PLGA)-icariin preserved visual function and RGC density after optic nerve infarct. The optic nerve edema, RGC apoptosis, and macrophage infiltration were inhibited by treatment with PLGA-icariin. The endogenous G-CSF expression switched the canonical NF-kB activation to the noncanonical NF-kB activation by promoting an alternative phosphorylation reaction of IKK-β. The noncanonical NF-κB activation further promoted the M2 microglia/macrophage polarization and AKT1 activation, which prevented the neuroinflammation and RGC apoptosis Granulocyte colony-stimulating factor. ConclusionOur study concluded that the protective mechanism of icariin is a G-CSF-induced non-canonical NF-kB activation, which may provide a potential therapeutic strategy for a patient with neuroinflammation diseases.