Potentiating effect and mechanism of extract of Jingfang Granules on activation of macrophages / 中国中药杂志

This study aimed to explore the potentiating effect and mechanism of the extract of Jingfang Granules(JFG) on the activation of macrophages. The RAW264.7 cells were treated with JFG extract and then stimulated by multiple agents. Subsequently, mRNA was extracted, and reverse transcription-polymerase chain reaction(RT-PCR) was used to measure the mRNA transcription of multiple cytokines in RAW264.7 cells. The levels of cytokines in the cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA). In addition, the intracellular proteins were extracted and the activation of signaling pathways was determined by Western blot. The results showed that JFG extract alone could not promote or slightly promote the mRNA transcription of TNF-α, IL-6, IL-1β, MIP-1α, MCP-1, CCL5, IP-10, and IFN-β, and significantly enhance the mRNA transcription of these cytokines in RAW264.7 cells induced by R848 and CpG in a dose-dependent manner. Furthermore, JFG extract also potentiated the secretion of TNF-α, IL-6, MCP-1, and IFN-β by RAW264.7 cells stimulated with R848 and CpG. As revealed by mechanism analysis, JFG extract enhanced the phosphorylation of p38, ERK1/2, IRF3, STAT1, and STAT3 in RAW264.7 cells induced by CpG. The findings of this study indicate that JFG extract can selectively potentiate the activation of macrophages induced by R848 and CpG, which may be attributed to the promotion of the activation of MAPKs, IRF3, and STAT1/3 signaling pathways.

Molecular identification of Tibetan medicinal plants based on ITS2 sequence / 中草药

Objective: The barcoded ITS2 DNA sequence was used to identify 44 Tibetan medicinal plants. Methods: Genomic DNA of Tibetan medicinal plants were extracted with high salt and low pH method. The PCR technique was performed to obtain the ITS2. A total of 145 ITS2 sequences was obtained belonging to 24 families, 39 genera, and 44 species. Some homologous sequences were also selected according to sequence alignment from Genbank database. The ITS2 sequences were aligned using Bioedit and the intraspecific and interspecific Kimura 2-parameter genetic distance was calculated using MEGA, and the neighbor-joining (NJ) phylogenetic trees were constructed to analyze phylogenetic relationship. Results: ITS2 regions have significant intra-and inter-specific difference, phylogenetic analysis based on ITS2 regions concurred with the result of morphological classification, and it could also determine the phylogenetic relationship between species. In addition, the secondary structure of the ITS2 sequence of Tibetan medicinal plants is different, providing another method for species identification. Conclusion: ITS2 can be used as a very effective single-locus barcode in the identification and phylogenetic study of Tibetan medicinal plants. The barcoding technique provides a scientific baseline for the utilization of resources and conservation of Tibetan medicinal plants.