Protective Effect of Angelicae Sinensis Radix-Chuanxiong Rhizoma Medicated Serum Against H2O2-induced Oxidative Damage of PC12 Cells Based on Nrf2/ARE Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the protective effect and molecular mechanism of Angelicae Sinensis Radix-Chuanxiong Rhizoma medicated serum （ASRCRS） against oxidative damage of PC12 cells induced by H₂O₂. Method:Oxidative damage of PC12 cells was induced by H₂O₂<italic> in vitro</italic>， and intervention was performed in the low-， medium-， and high-dose ASRCRS groups with a final volume fraction of 15%. The cell viability was determined by methyl thiazolyl tetrazolium （MTT） assay. Cell morphology was observed by an inverted fluorescence microscope. The content of lactate dehydrogenase （LDH） and malondialdehyde （MDA）， the activity of superoxide dismutase （SOD）， and the distribution of reactive oxygen species （ROS） in the cell supernatant were detected by the kits. Cell apoptosis was detected by Annexin V-FITC/PI double staining. The protein expression levels of nuclear factor E₂-related factor 2 （Nrf2）， Kelch-like epichlorohydrin associated protein-1 （Keap1）， heme oxygenase-1 （HO-1）， and SOD1 were detected by Western blot. Result:Oxidative damage was induced by 300 μmol·L^-1 H₂O₂ for 24 hours. Compared with the normal group， the model group showed abnormal cell morphology， reduced cell viability （<italic>P</italic><0.01）， increased LDH and MDA （<italic>P</italic><0.01）， blunted SOD activity， elevated intracellular distribution of ROS， down-regulated protein expression of Nrf2， HO-1， and SOD1 （<italic>P</italic><0.05， <italic>P</italic><0.05）， and up-regulated protein expression of Keap1 （<italic>P</italic><0.01）. Compared with the model group， ASRCRS groups displayed improved cell morphology， increased cell viability， inhibited cell apoptosis， potentiated SOD activity （<italic>P</italic><0.01）， suppressed release of LDH （<italic>P</italic><0.01） and generation of ROS， decreased content of MDA （<italic>P</italic><0.01）， up-regulated protein expression of Nrf2， HO-1 and SOD1 （<italic>P</italic><0.05， <italic>P</italic><0.01）， and down-regulated protein expression of Keap1 （<italic>P</italic><0.01）. Conclusion:ASRCRS could protect PC12 cells from oxidative damage induced by H₂O₂ by up-regulating the expression of Nrf2 to activate the Nrf2/antioxidant response element （ARE） signaling pathway， enhancing the ability to resist oxidative damage， and inhibiting cell apoptosis.

Blockade of retinal neovascularization by inhibiting erythropoietin in a mouse model / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To establish the murine model of oxygen induced retinopathy ( OIR) and to evaluate the inhibition of retinal neovascularization by erythropoietin ( EPO) blockade. METHODS: Neonates of C57BL/6 mouse ( P7 ) were exposed to 75%±2% oxygen for 5d and return to normal air environment when 12d ( P12 ) to establish oxygen-induced retinal neovascularization model. The neonates were divided into groups, injected with 0. 5μL solution containing 25ng ( group A), 50ng ( group B), 250ng ( group C) of soluble erythropoietin receptor ( EPO-R) or PBS (group D) at P12, P14 and P16 in the right eye. On P17, the litters were sacrificed and their right eyes were enucleated and fixed with 4% paraformaldehyde, made pathological section. The number of breakthrough internal limiting membrane neovascular nuclei was counted with pathological retinal morphology, understanding theproliferative degree of retinal neovascularization. RESULTS: The pathological sections showed the neovascular cell nuclei which penetrating the inner limiting membrane in intravitreal EPO receptor injection group was reduced statistically than that in PBS injection group, the difference was statistically significant ( P various concentrations of EPO receptor group was statistically significant (P CONCLUSION: Intravitreal injection of soluble EPO receptor can block EPO and improve neovascularization. The new method is expected to become new treatment of ocular neovascular diseases.

Effect of serum containing Jinmaitong Capsule on rats' Schwann cell apoptosis induced by high glucose concentration / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of serum containing Jinmaitong Capsule (JMT) on apoptosis of Schwann cells (SCs) that are cultured in high glucose at the cellular and molecular levels.</p><p><b>METHODS</b>SCs were cultured in Dulbecco's modified Eagle's medium (control group), high glucose (50 mmol/L) medium supplemented with 20% rat serum (HG group), and 50 mmol/L glucose medium supplemented with serum containing JMT (JMT group). SC apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling kit. The expression of Bcl-2 and the caspase-3 p20 subunit in SCs were detected by realtime fluorogenic quantitative polymerase chain reaction and confocal laser scanning microscopy, respectively.</p><p><b>RESULTS</b>No apoptosis was detected in SCs that were cultured in the control group. The percentage of apoptosis of SCs cultured in the HG group was much higher than that in the control group. The apoptosis of SCs in the JMT group was lower than that in the HG group. Fluorescence intensity of Bcl-2 and the expression of Bcl-2 mRNA in SCs that were cultured in the HG group were much lower than those in the control group and much higher than those in the JMT group (P<0.01). The fluorescence intensity of caspase-3 p20 and the expression of caspase-3 p20 mRNA in SCs that were cultured in the HG group were much higher than those in the control group (P<0.01), and they were remarkably lower in the JMT group (P<0.01).</p><p><b>CONCLUSIONS</b>JMT effectively prevents SC apoptosis that is induced by high glucose. This effect may be because of increased expression of Bcl-2 mRNA and protein and decreased expression of caspase-3 p20 mRNA and protein.</p>

Effects of Jinmaitong capsule on oxidative stress and cell apoptosis of dorsal root ganglion in diabetic rats / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the effects of Jinmaitong capsule on oxidative stress and cell apoptosis of dorsal root ganglion (DRG) in rats with diabetic peripheral neuropathy.</p><p><b>METHODS</b>Sixty male SD rats were randomly divided into normal group and model groups. The diabetic rat models were established using Streptozotocin (STZ) method (60 mg/kg of intraperitoneal injection), and then randomly divided Jinmaitong low, middle, and high-dose groups and vitamin C group. All the experimental rats were sacrificed at 16-week and then the DRG was isolated. The morphological changes of DRG were observed using the Nissl's staining, and the NADPH oxidase subunit p22-phox, Cyt C, Bcl-2, and Caspase-3 of DRG in rats were detected by immunohistochemistry and quantitative reverse transcription PCR (qRT-PCR). Cell apoptosis was detected by TUNEL.</p><p><b>RESULTS</b>Compared with the model group, the expressions of NADPH oxidase subunit p22-phox protein, Cyt expression of C protein, Caspase-3 protein, and mRNA cell apoptosis rate in each treatment group significantly decreased whereas the expressions of Bcl-2 mRNA and protein significantly increased (P<0.05 or P<0.01). The Jinmaitong high-dose group had the best effect and was significantly different from that of the vitamin C group (P<0.01).</p><p><b>CONCLUSIONS</b>Jinmaitong capsule can prevent the nerve injury in rats with diabetic peripheral neuropathy by inhibiting oxidative stress and decreasing the apoptosis. The high-dose Jinmaitong capsule has the best effect and is superior to vitamin C.</p>