Action characteristics and mechanisms of Panlongqi Tablets in treating fractures based on research strategy integrating Meta-analysis,network pharmacology,and experimental verification

This study adopted a research strategy and method integrating Meta-analysis, network pharmacology, and experimental verification, aiming to clarify the clinical effect and action characteristics of Panlongqi Tablets in treating fractures and preliminarily reveal the related action mechanism. Firstly, 13 randomized controlled trials(RCTs) of Panlongqi Tablets for treating fractures that met the criteria were retrieved and included. The analysis results show that compared with the control group, Panlongqi Tablets could significantly improve the imaging manifestations and motor function of patients, increase the total effective rate, reduce the swelling score and coagulation indicators, and increase the osteogenic factors, thereby promoting fracture healing. Secondly, the differential gene set of rat fractures at different stages(seven, 14, and 28 days) in the GEO database, namely GSE262132, was collected, and the drug target spectra of Panlongqi Tablets were obtained from databases such as TCMSP. Through the correspondence evaluation for the " disease at different stages and prescription" by using Jaccard and cosine similarity, it was found that the best correspondence was at seven days(early stage). Through the construction of the " differential genes at different fracture stages and candidate targets of Panlongqi Tablets" interaction network for functional enrichment analysis, it was found that the core targets in each stage were enriched in signaling pathways regulating inflammation immunity, pain, blood circulation, and bone metabolism, among which the correlation was the strongest as the phosphatidylinositol-3-kinase(PI3K)-protein kinase B(Akt) signaling pathway. The mechanism of the above candidate signaling molecules was verified by using a rat model of tibial transverse fracture. Western blot was used to detect the expression levels of PI3K, Akt, phosphorylated protein kinase B(p-Akt), and vascular endothelial growth factor A(VEGFA), and ELISA was used to detect the expression levels of transforming growth factor β(TGF-β), bone morphogenetic protein-2(BMP-2), bone alkaline phosphatase(BALP), type I procollagen amino-terminal propeptide(PINP), and osteocalcin(OCN). The results show that Panlongqi Tablets could significantly increase the protein expression levels of PI3K, p-Akt, Akt, and VEGFA in the healing tissue and could significantly elevate the contents of TGF-β, BMP-2, BALP, PINP, and OCN in the serum, indicating that the mechanism of Panlongqi Tablets promoting fracture healing may be related to the activation of the PI3K-Akt signaling pathway. The research results are helpful to clarify the efficacy connotation of the " Qi yao" and guide the rational clinical medication of Panlongqi Tablets.