Effect of Guipitang on ERK1/2 and p38 MAPK in Rats with Myocardial Ischemia / 中国实验方剂学杂志

ObjectiveTo explore the therapeutic effect and mechanism of Guipitang on rats with myocardial ischemia. MethodFifty SD rats were divided into five groups： a control group， a model group， low and high-dose Guipitang （7.52， 15.04 g·kg-1） groups， and a trimetazidine group （0.002 g·kg-1）. By intragastric administration of vitamin D3 and feeding rats with high-fat forage and injecting isoproterenol， the rat model of myocardial ischemia was established. After drug treatment of 15 d， an electrocardiogram （ECG） was performed to analyze the degree of myocardial injury. A fully automatic biochemical analyzer was used to detect the changes in the serum levels of total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C）. Hematoxylin-eosin （HE） staining and Masson staining were used to observe myocardial histopathological changes. TdT-mediated dUTP nick end labeling （TUNEL） staining was used to detect cardiomyocyte apoptosis. Western blot was adopted to detect the protein levels of extracellular signal-regulated kinase 1/2 （ERK1/2）， phospho-ERK1/2 （p-ERK1/2）， p38 mitogen-activated protein kinase （p38 MAPK）， phospho-p38 MAPK （p-p38 MAPK）， B-cell lymphoma-2 （Bcl-2）-associated X （Bax）， Bcl-2， and cleaved cysteine aspartate proteolytic enzyme （cleaved Caspase-3）. ResultCompared with the control group， the ECG S-T segment decreased in the model group. The serum levels of TC， TG， and LDL-C were increased significantly （P<0.05）. The arrangement of myocardial tissue was disordered， and the proportion of cardiomyocyte apoptosis increased. The protein levels of cleaved Caspase-3， Bax， and p-p38 MAPK in the heart were increased， and the Bcl-2 expression was decreased （P<0.05）. Compared with the model group， the S-T segment downward shift was restored in the low and high-dose Guipitang groups and trimetazidine group， and the levels of TC， TG， and LDL-C were decreased. The protein expression of cleaved Caspase-3 and Bax in the heart dropped， and p-p38 MAPK and p-ERK1/2 protein expressions increased significantly （P<0.05）. The degree of myocardial injury was alleviated， and the proportion of cardiomyocyte apoptosis decreased. Bcl-2 protein expression was increased significantly in the low-dose Guipitang group （P<0.05）. ERK1/2 and p38 MAPK proteins had no significant difference among different groups. ConclusionGuipitang could alleviate myocardial injury and inhibit cardiomyocyte apoptosis in rats by activating the expression of ERK1/2 and p38 MAPK.

Effect of Wenjing Tongluo Decoction on VEGF/VEGFR2/ERK1/2 Signaling Pathway in Mice with Knee Osteoarthritis / 中国实验方剂学杂志

Objective:To investigate the possible mechanism of Wenjing Tongluo decoction （WTD） in alleviating articular cartilage defect in knee osteoarthritis （KOA） and delaying joint degeneration. Method:The KOA model was established by anterior cruciate ligament transection （ACLT）. Mice were classified into sham-operated group， model group， WTD high-dose and low-dose groups， and positive control group. Four weeks after modeling， WTD groups and the positive control group were given WTD （80， 20 g·kg^-1） and glucosamine sulfate capsules （0.29 g·kg^-1）， respectively， and the sham-operated group and model group received normal saline of the equivalent volume. After continuous intervention for 4 weeks， hemoxylin-eosin （HE） staining was used to observe the morphological changes of cartilage and Mankin scoring system was employed to score the knee cartilage. Western blot was combined with Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） to detect the protein and mRNA levels of vascular endothelial growth factor <italic>α</italic> （VEGFA）， vascular endothelial growth factor receptor 2 （VEGFR2）， extracellular signal-related kinase 1/2 （ERK1/2） and a disintegrin and metalloproteinase with thrombospondin motifs 4 （ADAMTS4）. Result:The Mankin score in the model group increased as compared with that in the sham-operated group （<italic>P</italic><0.01）. Compared with the model group， administration groups demonstrated alleviated articular cartilage defect and low Mankin score （<italic>P</italic><0.01）， but there was no statistical significance in Mankin score between the WTD groups and positive control group. The protein and mRNA levels of VEGFA， VEGFR2， ERK1/2， and ADAMTS4 in the model group were significantly higher than those in the sham-operated group （<italic>P</italic><0.01）. The protein expression of VEGFA and ERK1/2 was inhibited in each administration group as compared with that in the model group （<italic>P</italic><0.01）， and the inhibition in the positive control group was stronger than that in the WTD low-dose group （<italic>P</italic><0.05） but weaker than that in the WTD high-dose group （<italic>P</italic><0.01）. Glucosamine Sulfate capsules suppressed the expression of VEGFR2 and ADAMTS4 to the extent the same with low-dose WTD but weaker than the high-dose WTD （<italic>P</italic><0.05）. Conclusion:WTD can relieve the articular cartilage injury in KOA mice， and the mechanism may be related to VEGF/VEGFR2/ERK1/2 signaling pathway.

Study on Mechanism and Validation of Effect of Astragali Radix in Treatment of Parkinson's Disease by Network Pharmacology / 中国实验方剂学杂志

Objective:To analyze the mechanism of Astragali Radix in the treatment of Parkinson's disease by network pharmacology and PC-12 extracellular model. Method:Traditional Chinese medicines systems pharmacology platform （TCMSP） and CD-HIT databases were used to screen out active components and targets of Astragali Radix， GENECARDS and Online Mendelian Inheritance in Man （OMIM） databases were used to screen out targets relating to Parkinson's disease and draw component-target network， STRING database was used to build the protein-protein interaction network， Bioconductor Cluster Profiler was applied in Gene Ontology （GO） enrichment analysis， and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis. PC-12 cells were pretreated with water extract of Astragali Radix， and Western blot was used to assess the expression of phosphorylation extracellular regulatory protein kinase 1/2（p-ERK1/2）， ERK1/2， B cell lymphoma -2 （Bcl-2） associated X protein （Bax）， Bcl-2， cysteine aspartic acid protease -3 （Caspase-3） and cleaved-Caspase3 （c-Caspase-3）. The levels of interleukin-6 （IL-6）， IL-10 and tumor necrosis factor-α（TNF-α） were determined by enzyme-linked immunosorbent assay （ELISA） interleukin-6 （IL-6）， IL-10， tumor necrosis factor-α（TNF-α）. Result:Network pharmacology showed that 14 compounds in Astragali Radix， including emodin and quercetin， played a role through multi-target and multi-channel synergy， involving ERK signal pathway， Bax， Bcl-2， Caspase-3， IL-6， IL-10， TNF-α and other target proteins. Western blot showed that the expressions of Bax， p-ERK1/2， c-Caspase-3 and Bcl-2 in Astragali Radix Extract group decreased， while the expressions of Bcl-2 in Astragali Radix Extract group was significantly higher than that in model group （P<0.05）. There were statistical differences between the two groups. ELISA showed that Astragali Radix water extract could reduce the expressions of IL-6， TNF-α， but increase the expression of IL-10， with statistical differences from the model group （P<0.05）. Conclusion:This study shows that Astragali Radix can affect the expressions of these proteins， and verify the results of network pharmacology， so as to provide a basis for further study of Astragali Radix in the treatment of Parkinson's disease.