A Novel Biochemical Study of Anti-Dermal Fibroblast Replicative Senescence Potential of Panax Notoginseng Oligosaccharides

Abstract

Dermal fibroblast replicative senescence that often occurs in aging skin is characterized by loss of cell proliferative capacity, cell cycle arrest, decreased cell elongation, and decreased synthesis of dermal extracellular matrix (ECM) components. Although Panax notoginseng is known for its effectiveness in alleviating many age-related degenerative diseases, few studies have evaluated P. notoginseng components for efficacy or mechanisms of action in delaying cell replicative senescence. In this study, P. notoginseng oligosaccharides (PNO) were isolated using a stepwise purification procedure involving water extraction and alcohol precipitation followed by DEAE Sepharose Fast Flow column chromatography, preparative high performance liquid chromatography, and size-exclusion chromatography. Monosaccharides detected in PNO constituents included mannose, galactose, and sorbitose in relative molar proportions of 14.2:12.3:1, respectively, aligning with PNO absorption spectrum results resembling typical known spectra for sugars. In vitro, PNO treatment of replicative senescent NIH-3T3 fibroblasts significantly promoted cell vitality, inhibited SA-β-galactosidase (SA-β-Gal) activity, and reduced p16 and p21 protein-level expression. Moreover, PNO treatment of senescent fibroblasts led to a lower proportion of G1 phase cells and higher proportion of S phase cells, while also inducing aging NIH-3T3 cells to migrate and synthesize collagen-I (CoL-I). Mechanistically, PNO treatment up-regulated expression of proliferating cell nuclear antigen (PCNA), cyclin E, cyclin D1, and cyclin-dependent kinase 4 (CDK4) proteins and promoted phosphorylation of MEK, p38, and ERK1/2 to trigger cell cycle progression. Additionally, PNO treatment also up-regulated protein-level expression of TGF-β1 and levels of p-Smad2/3, p-FAK, and p-Pax to trigger CoL-I synthesis and cell migration. Taken together, these findings demonstrate that oligosaccharides purified from P. notoginseng could reverse fibroblast replicative senescence by promoting fibroblast cell proliferation, migration, and CoL-I production.

Protective Effect of Panax notoginseng Extract Fermented by Four Different Saccharomyces cerevisiae Strains on H2O2 Induced Oxidative Stress in Skin Fibroblasts

“…Zhai et al found that Panax notoginseng oligosaccharides isolated from Panax notoginseng could reverse fibroblast replicative senescence by promoting fibroblast proliferation, migration and CoL-I production. 35 Panax notoginseng saponins can protect tumor necrosis factor-α-induced osteoarthritis rat chondrocytes from aging and apoptosis by regulating the PI3K-AKT-mTOR pathway, thereby delaying the destruction of cartilage by osteoarthritis. 36 Moreover, Panax notoginseng polysaccharides can significantly prolong the average lifespan of wildtype Caenorhabditis elegans and may increase the survival rate of wildtype Caenorhabditis elegans under heat stress by increasing the activities of SOD and CAT and reducing the content of MDA.…”

Ginsenosides improve reproductive capability of aged female Drosophila through mechanism dependent on ecdysteroid receptor (ECR) and steroid signaling pathway

“…Kr-h1 (Santa Cruz Biotechnology, Santa Cruz, USA). Next, the membranes were incubated with appropriate secondary antibodies (Li-Cor, Lincoln, NE, USA) for 1 h at room temperature ( 31 ). Finally, chemiluminescence-based visualization and analysis of protein bands were conducted using a FluorChem imaging system (ProteinSimple, San Jose, CA, USA).…”

Effects of Carissa carandas Linn. Fruit, Pulp, Leaf, and Seed on Oxidation, Inflammation, Tyrosinase, Matrix Metalloproteinase, Elastase, and Hyaluronidase Inhibition

“…Besides, fibroblasts are present throughout the matrix and connective tissue of the body. Therefore, NIH-3T3 cell line has been previously used for studying the regulation of fibrillar collagen synthesis, cellular cytotoxicity, genotoxicity, inhibition of UV-induced skin photoaging, inhibition of MMPs, etc., of tested compounds or topical formulations [ 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 ]. Briefly, NIH/3T3 cell line were cultured in DMEM culture medium with 10.0% newborn calf serum, and 1.0% antibiotic–antimycotic (100×).…”