UPLC-Q-TOF-MS Reveals Mechanisms of Modified Qing'e Formula in Delaying Skin Photoaging and Regulating Circadian Rhythm
Objective:To reveal the active substances and mechanisms of modified Qing'e formula(MQEF)in delaying skin photoaging by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS),network pharmacology,and cell experiments.Methods:UPLC-Q-TOF-MS and a literature review were employed to analyze the transdermally absorbed components in mice after the topical application of MQEF.The potential targets of MQEF in treating skin photoaging were retrieved from databases.The compound-potential target network and protein-protein interaction network were constructed to screen the key components and core targets.A photoaging cell model was established by irradiating HaCaT cells with medium-wave ultraviolet B(UVB).The safe doses of bakuchiol(BAK)and salvianolic acid B(SAB)for treating HaCaT cells and the effects of BAK and SAB on the viability of cells exposed to UVB irradiation were determined by the cell counting kit-8(CCK-8)method.The reactive oxygen species(ROS)fluorescent probe was used to measure the ROS production in the cells treated with BAK and SAB.The expression levels of genes related to oxidative stress,inflammation,collagen metabolism,and circadian rhythm clock were measured by Real-time PCR.Results:A total of 24 transdermally absorbed components of MQEF were identified,which acted on 367 potential targets,and 417 targets related to skin photoaging were screened out,among which 47 common targets were predicted as the targets of MQEF in treating skin photoaging.MQEF exerted the anti-photoaging effect via key components such as BAK and SAB,which acted on core proteins such as serine/threonine kinase 1(Aktl)and mitogen-activated protein kinase 3(MAPK3)and intervened in core pathways such as the tumor necrosis factor(TNF)and phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt)signaling pathways.Compared with the model group,the administration of BAK and SAB increased the survival rate of HaCaT cells(P＜0.01),down-regulated the mRNA levels of cyclooxygenase-2(COX-2),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),matrix metalloproteinase-1(MMP-1),and matrix metalloproteinase-9(MMP-9)(P＜0.01),and up-regulated the mRNA levels of heme oxygenase-1(HO-1)and NAD(P)H quinone dehydrogenase 1(NQO-1)(P＜0.05,P＜0.01)in photoaged HaCaT cells.In addition,it eliminated excess ROS production induced by UVB and up-regulated the mRNA levels of brain and muscle ARNT-like 1(BMAL1)and circadian locomotor output cycles kaput(CLOCK)associated with circadian clock(P＜0.05,P＜0.01).Conclusion:MQEF delays skin photoaging through the coordinated effects of various components,multiple targets,and diverse pathways.The key components BAK and SAB in MQEF exhibit anti-photoaging properties,which involve inhibiting oxidative stress,preventing collagen degradation,mitigating inflammation,and maintaining normal skin circadian rhythms by regulating clock gene expression.
modified Qing'e formula(MQEF)skin photoagingcircadian rhythmultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS)network pharmacology
万方数据
维普
