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.

Protective Effect of Ferulic Acid on PC12 Cells with H2O2-induced Oxidative Damage / 中国实验方剂学杂志

Objective:To investigate the protective effect of ferulic acid on PC12 cells injured by H2O2 and the molecular mechanisms. Method:The oxidative stress model was established by treating PC12 cells with H2O2, and then different dosages of ferulic acid (1, 10, 100 μmol·L-1) were used for intervention. Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the cell viability,lactate dehydrogenase (LDH) and malondialdelyde (MDA) in cell supernatant, and superoxidedismutase (SOD) in cells was tested by biochemical method respectively. Insulin-like growth factors-1 (IGF-1) mRNA and protein expressions were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot. Result:The 24 h intervention with different dosages of ferulic acid (1, 10, 100 μmol·L-1) could significantly improve the oxidative damage of PC12 cells induced by H2O2, compared with the model group, ferulic acid at 1,10,100 μmol·L-1 significantly increased PC12 cells viability,significantly decreased LDH and MDA content in cell supernatant (PPPPPConclusion:Ferulic acid exerts a protective effect on H2O2-inducing PC12 cells injury,which might be related to insulin signaling pathways.

Study on diagnosis criteria of fire-heat syndrome based on receiver operating characteristic curve and principal component analysis / 中国结合医学杂志

<p><b>OBJECTIVE</b>To establish the diagnostic quantitative criteria for fire-heat syndrome (FHS) of Chinese medicine (CM) based on the receiver operating characteristic (ROC) curve and principal component analysis (PCA).</p><p><b>METHODS</b>The symptoms and signs of FHS cases and healthy subjects from Guangzhou, Henan and Hunan of China were collected through questionnaire, and the diagnostic quantitative score tables were established for the three regions, respectively, with the method of maximum likelihood analysis. The homogeneity test was then performed on the diagnostic score tables for the three regions with ROC curve, and the diagnostic efficiency of diagnostic score tables for the three regions was compared with the prospective test and retrospective test. The method of PCA was adopted to obtain the analysis matrix for classifying the tapes of FHS.</p><p><b>RESULTS</b>Twenty-seven elements of FHS were confirmed through Chi-square test, and the diagnostic score tables for the three regions were established with the method of maximum likelihood analysis on the basis of the collected case data. According to the ROC curve test, the areas under ROC curve of Guangzhou diagnostic score table assessment with candidates in Guangzhou, Henan and Hunan were 0.998, 0.961 and 0.956, respectively. It showed that the diagnostic efficiency of Guangzhou diagnostic score tables was the highest one. With the prospective test, the area under ROC of Guangzhou diagnostic score table was 0.949, and more than any other diagnostic score table. By PCA, FHS was classified into excess fire and deficiency fire, and then classified into syndrome of flaring up of Heart (Xin) fire, syndrome of Lung (Fei)-Stomach (Wei) excess fire, syndrome of deficiency of Liver (Gan)-yin and Kidney (Shen)-yin, and syndrome of deficiency of Lung-yin from the view of viscera. In the retrospective test, the consistency with clinicians' diagnosis was 69.4%, and in the prospective test, it was 70.1%.</p><p><b>CONCLUSIONS</b>The Guangzhou diagnostic score table could be used as the recommended criteria for the diagnosis of FHS. The classification of FHS was basically in conformity with the clinical situation.</p>