RNA Sequencing Analysis and Verification of Paeonia ostii 'Fengdan' CuZn Superoxide Dismutase (PoSOD) Genes in Root Development.

Tree peony (Paeonia suffruticosa) is a significant medicinal plant. However, the low rooting number is a bottleneck problem in the micropropagation protocols of P. ostii 'Fengdan'. The activity of superoxide dismutase (SOD) is closely related to root development. But research on the SOD gene's impact on rooting is still lacking. In this study, RNA sequencing (RNA-seq) was used to analyze the four crucial stages of root development in P. ostii 'Fengdan' seedlings, including the early root primordium formation stage (Gmfq), root primordium formation stage (Gmf), root protrusion stage (Gtq), and root outgrowth stage (Gzc). A total of 141.77 GB of data were obtained; 71,718, 29,804, and 24,712 differentially expressed genes (DEGs) were identified in the comparison groups of Gmfq vs. Gmf, Gmf vs. Gtq, and Gtq vs. Gzc, respectively. Among the 20 most highly expressed DEGs in the three comparison groups, only the CuZnSOD gene (SUB13202229, PoSOD) was found to be significantly expressed in Gtq vs. Gzc. The overexpression of PoSOD increased the number of adventitious roots and promoted the activities of peroxidase (POD) and SOD in P. ostii 'Fengdan'. The gene ADVENTITIOUS ROOTING RELATED OXYGENASE1 (PoARRO-1), which is closely associated with the development of adventitious roots, was also significantly upregulated in overexpressing PoSOD plants. Furthermore, PoSOD interacted with PoARRO-1 in yeast two-hybrid (Y2H) and biomolecular luminescence complementation (BiFC) assays. In conclusion, PoSOD could interact with PoARRO-1 and enhance the root development of tube plantlets in P. ostii 'Fengdan'. This study will help us to preliminarily understand the molecular mechanism of adventitious root formation and improve the root quality of tree peony and other medicinal plants.

Meta-Analysis of Preventive and Therapeutic Effects of Ligustrazine on Airway Remodeling in Asthmatic Rats.

Objective: To assess the effect of ligustrazine on airway remodeling in asthmatic rats. Methods: To collect studies on the effects of ligustrazine on airway remodeling in asthmatic rat models, PubMed, Embase, CBM, Cochrane, Chinese Knowledge Infrastructure (CNKI), VIP, and Wanfang data (WANFANG) were searched using a computer. Two investigators independently screened the literature, extracted the data, and assessed the methodological quality by complying with the inclusion criteria. Moreover, a meta-analysis was conducted by adopting Stata 11.0. Results: On the whole, 10 articles were included. As indicated from the meta-analysis, we have the following: ① ligustrazine was capable of reducing the thickness of the airway smooth muscle and inhibiting the proliferation of smooth muscle (WMD = -5.98, 95% CI (-7.75, -4.42), P ≤ 0.001); ② ligustrazine could reduce the thickness of the airway wall and mitigate tracheal stenosis (WMD = 0.12, 95% CI (0.05, 0.20), P ≤ 0.001); ③ ligustrazine could decrease the number of eosinophils in the lung tissue and reduce airway inflammation (WMD = -14.47, 95% CI (-18.09, -10.86), P ≤ 0.001). Conclusion: Ligustrazine was demonstrated to be an effective therapeutic drug in asthmatic rats by preventing and treating airway remodeling. Further high-quality experimental studies should be conducted to investigate the mechanism of ligustrazine action in depth.

Mechanism of Tetrandrine Against Endometrial Cancer Based on Network Pharmacology.

BACKGROUND: Endometrial cancer (EC) is one of the most common gynaecological malignancies, and its incidence has been rising over the past decade. Tetrandrine, a bisbenzylisoquinoline alkaloid, has been isolated from a vine used in traditional Chinese medicine, Stephania tetrandra. However, the key mechanism of tetrandrine in EC is still unclear. PURPOSE: This research was designed to predict the molecular mechanisms of tetrandrine against EC based on network pharmacology and to further verify these predictions by in vitro experiments. METHODS: The potential therapeutic targets of tetrandrine against EC were predicted by using public databases. Afterwards, the protein-protein interaction (PPI) network of the common targets was constructed, and the key gene targets were obtained. Biological function and pathway enrichment analyses were performed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Furthermore, molecular docking and in vitro experiments were carried out to verify the predictions. The cell counting kit­8 (CCK­8) assay, Hoechst 33258 staining, flow cytometry analysis, qRT-PCR, Western blot analysis and an immunofluorescence assay were performed. RESULTS: Our findings identified 111 potential therapeutic targets of tetrandrine against EC. We obtained 7 key gene targets from the PPI network analysis. Furthermore, GO enrichment analysis indicated that these targets were mainly associated with metabolic processes, responses to stimulus, and biological regulation. The KEGG pathway analysis showed that the common targets were mainly distributed in the PI3K/Akt signalling pathway. A potential interaction of tetrandrine with Akt1 was revealed by molecular docking. In addition, in vitro experiments showed that tetrandrine significantly inhibited cell proliferation and induced apoptosis in Ishikawa and HEC-1-B cells in dose- and time-dependent manners. The results also revealed that tetrandrine can downregulate the expression of Bcl-2 and upregulate the expression of Bax at the mRNA level. The mRNA levels of Akt were not significantly different in the various tetrandrine (0, 10 and 20µM) groups. However, Western blot analysis demonstrated that the protein expression ratios of p-Akt/Akt decreased at the protein level. The results were further confirmed by immunofluorescence assays. CONCLUSION: Based on bioinformatic analysis and experimental verification, our findings demonstrated that tetrandrine exerted tumour-suppressive effects on EC by regulating the PI3K/Akt signalling pathway.