Sanggenol P activates HIF-1α to delay skin aging-raw data

It mainly includes high-throughput screening, cell function experiments, protein expression detection, and molecular mechanism analysis, among other types of data. The high-throughput screening part used a HEK293 cell model stably transfected with the HRE luciferase reporter gene (pGL4.42 [luc2P/HRE/Hygro]) to screen for HIF signal activation activity from a compound library containing 439 natural product small molecules.

The processing time was 24 hours, and the luciferase activity was detected. The cell experiment data mainly comes from human immortalized keratinocytes (HaCaT) and human skin fibroblasts (HSF), and H ₂ O ₂ - induced oxidative stress and cell aging models were established, with SP treatment concentration ranging from 0-40 μ M and treatment time of 24-48 h. The detection indicators include cell viability (MTT method), oxidative stress indicators (MDA, SOD, GSH Px), inflammatory factors (TNF - α, IL-6), aging related indicators (SA - β - gal), as well as extracellular matrix related genes (MMP-1, COL1A1, COL3A1) and tight junction protein ZO-1 expression. The protein expression data was obtained from Western blot experiments, mainly detecting the expression changes of HIF-1 α and HIF-2 α at different treatment times (6, 12, 24 h); The gene expression data was obtained through RT qPCR and subjected to relative quantitative analysis using the 2 °Δ Ct method.