Study On Synergistic Mechanism Of Scutellaria Baicalensis Fractions On FGF1 Promoting Wound Healing

Objective:In view of the research on the synergistic effect of the active ingredients of traditional Chinese medicine,based on multi-compartment electrophoresis screening technology,the active components that have a synergistic effect with FGF1 to promote wound healing were screened from Scutellaria baicalensis Georgi,and explore the synergistic mechanism of the components of Scutellaria baicalensis Georgi and FGF1.Method:1.Optimize multi-compartment electrophoresis screening technical parameters and screen the components of Scutellaria baicalensis Georgi that synergistically with FGF1:optimize multi-compartment electrophoresis screening technical parameters(p H,time,voltage);use FGF1 as the target protein to screen synergistic components;establish LC-MS /MS method detects component qualitative and quantitative analysis.The Molecular Operating Environment 2019 was used to simulate the molecular docking of Scutellaria baicalensis Georgi components and FGF1 to verify the binding mode of the selected Scutellaria baicalensis Georgi components and FGF1.2.Exploring the synergistic effect of FGF1-Scutellaria baicalensis Georgi component: after combined administration of FGF1-Scutellaria baicalensis Georgi components,the effects of NIH3T3 cell proliferation and cell migration were investigated.ELISA kit was used to detect collagen content and the synergistic effect of FGF1-Scutellaria baicalensis Georgi components was investigated.3.Based on the PI3K/AKT/mTOR signal pathway to explore the molecular mechanism of the Scutellaria baicalensis Georgi component-FGF1 synergy of Scutellaria baicalensis Georgi: extract sample RNA,and use q RT-PCR technology to detect the relative relationship of PI3 K,AKT,and mTOR mRNA genes in the PI3K/AKT/mTOR signal pathway the amount of expression.Western blot was used to detect the relative expression of PI3 K,p-PI3 K,AKT,p-AKT,mTOR and p-mTOR proteins in the PI3K/AKT/mTOR signaling pathway.Results:1.Multi-compartment electrophoresis screening technical parameters: Scutellaria baicalensis Georgi extract and FGF1 target protein,incubated at 37 ℃ for 30 min,10 mM ammonium acetate p H 4,electrophoresis voltage 14 V,electrophoresis time 15 min,target protein FGF1 mobility 60.58%(RSD=1.825%).2.Based on the multi-compartment electrophoresis screening technology,three components were screened from Scutellaria baicalensis Georgi,and the three active components were qualitatively identified by LC-MS/MS: Baicalin and Oroxylin A-7-O-β-D-glucuronide,Wogonoside.At the same time,the MRM mode was used to quantify the three components,and the molar ratio was determined to be 2:2:1.3.After the Molecular Operating Environment 2019 software is docked,they are all combined with FGF1,and the bonding method is hydrogen bonding.4.When FGF1 is 2.5 nM,the effect of promoting cell proliferation is the best;the combined administration of FGF1 and Scutellaria baicalensis Georgi component can promote cell proliferation,which is significantly different from the FGF1 group(P<0.001);and can increase collagen The synthesis of FGF1 and 3STD is significantly different from that of FGF1 and 3STD alone(P<0.001);it can also increase the migration rate of cells,which is significantly different from FGF1 and 3STD alone(P<0.001).5.The combined administration of FGF1 and Scutellaria baicalensis Georgi can activate the relative expression of PI3 K,AKT and mTOR mRNA genes and proteins in the PI3K/AKT/mTOR signaling pathway in cells.Conclusion:Using multi-compartment electrophoresis screening technology,three active components(Baicalin,Oroxylin A-7-O-β-D-glucuronide and Wogonoside)that act synergistically with the target protein FGF1 were screened from Scutellaria baicalensis Georgi.The molar ratio is 2:2:1.After combined administration,it synergistically promotes the proliferation and migration of NIH3T3 cells,and increases the synthesis of cell collagen.At the same time,it activates the PI3K/AKT/mTOR signaling pathway and regulates the expression of PI3 K,AKT,mTOR genes and proteins.The preliminary coordination mechanism between FGF1 and Scutellaria baicalensis Georgi may be related to the activation of PI3K/AKT/mTOR signaling pathway.