Journal: Scientific Reports

Article Title: Polysaccharide from Prinsepia utilis Royle maintains the skin barrier by mediating differentiation, lipid metabolism and tight junction of keratinocyte

doi: 10.1038/s41598-025-01960-w

Figure Lengend Snippet: PUR-P contributes to the expression of proteins related to differentiation, lipid metabolism and tight junctions in keratinocytes. ( A – E ) After 48 and 72 h of treatment with 10 mg/ml PUR-P, the expression of differentiation-associated (K1 and K10; A ), proliferation-associated (K16 and K17; A), sebum-synthesis-associated (CERS3, FAS, and HMGCS2; B ), tight junction-associated (ZO-1, Occludin, and Claudin-1; C ), cornified envelope-associated (FLG, INV, and LOR; D ), and PPAR-associated (PPARα, PPARβ/δ, and PPARγ; E ) proteins were identified using western blot assay. The original gel blot is shown in the Supplementary Information file. DRPs: keratinocyte differentiation-related proteins; PRPs: keratinocyte proliferation-related proteins; SSRPs: sebum synthesis-related proteins; TJRPs: tight junction-related proteins; CEPs: cornified envelope-related proteins; PPRPs: PPAR-related proteins. N = 3; One-way ANOVA test with Tukey’s multiple comparisons test or Kruskal–Wallis test with Dunn’s multiple comparisons test; Statistical significance was defined as P < 0.05.

Article Snippet: The KD-NC and KD-PPARα groups were infected with sh-NC and sh-PPARα_1 lentivirus using the HitransG P (REVG005, Genechem, China) for 48 h. The Norathyriol group were treated with 100 μM Norathyriol, a PPARα antagonist (HY-N1029, MCE, USA) for 24 h. Subsequently, the PUR-P, Norathyriol, KD-NC, and KD-PPARα groups were treated with 10 mg/ml PUR-P for 48 and 72 h. The Solvent group were treated with equal volumes of sterile water.

Techniques: Expressing, Western Blot

Journal: Scientific Reports

Article Title: Polysaccharide from Prinsepia utilis Royle maintains the skin barrier by mediating differentiation, lipid metabolism and tight junction of keratinocyte

doi: 10.1038/s41598-025-01960-w

Figure Lengend Snippet: PUR-P facilitates the expression of mRNAs related to differentiation, lipid metabolism and tight junctions in keratinocytes. ( A – D ) PUR-P promotes the expression of differentiation-associated (K1 and K10; A ), sebum synthesis-associated (CERS3 and FAS; B ), tight junction-associated (ZO-1; C ), and PPAR-associated (PPARα and PPARβ/δ; D ) mRNAs in keratinocytes. DRPs: keratinocyte differentiation-related proteins; PRPs: keratinocyte proliferation-related proteins; SSRPs: sebum synthesis-related proteins; TJRPs: tight junction-related proteins; PPRPs: PPAR-related proteins. N = 3; One-way ANOVA test with Tukey’s multiple comparisons test or Kruskal–Wallis test with Dunn’s multiple comparisons test; Statistical significance was defined as P < 0.05.

Article Snippet: The KD-NC and KD-PPARα groups were infected with sh-NC and sh-PPARα_1 lentivirus using the HitransG P (REVG005, Genechem, China) for 48 h. The Norathyriol group were treated with 100 μM Norathyriol, a PPARα antagonist (HY-N1029, MCE, USA) for 24 h. Subsequently, the PUR-P, Norathyriol, KD-NC, and KD-PPARα groups were treated with 10 mg/ml PUR-P for 48 and 72 h. The Solvent group were treated with equal volumes of sterile water.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Polysaccharide from Prinsepia utilis Royle maintains the skin barrier by mediating differentiation, lipid metabolism and tight junction of keratinocyte

doi: 10.1038/s41598-025-01960-w

Figure Lengend Snippet: Norathyriol and PPARα shRNA lentivirus alleviate PUR-P-induced upregulation of PPARα expression. ( A , B ) Western blot ( A ) and qRT-PCR ( B ) assays characterized the effect of shRNA lentivirus on the expression of PPARα protein and mRNA in keratinocytes. ( C , D ) Representative images of PPARα gel blot ( C ) and PPARα mRNA expression in different groups ( D ). PUR-P facilitates the expression of PPARα protein and mRNA in keratinocytes, and this is limited by Norathyriol (PPARα antagonist) and PPARα shRNA lentivirus. The original gel blot is shown in the Supplementary Information file. N = 3; One-way ANOVA test with Tukey’s multiple comparisons test; Statistical significance was defined as P < 0.05.

Article Snippet: The KD-NC and KD-PPARα groups were infected with sh-NC and sh-PPARα_1 lentivirus using the HitransG P (REVG005, Genechem, China) for 48 h. The Norathyriol group were treated with 100 μM Norathyriol, a PPARα antagonist (HY-N1029, MCE, USA) for 24 h. Subsequently, the PUR-P, Norathyriol, KD-NC, and KD-PPARα groups were treated with 10 mg/ml PUR-P for 48 and 72 h. The Solvent group were treated with equal volumes of sterile water.

Techniques: shRNA, Expressing, Western Blot, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Polysaccharide from Prinsepia utilis Royle maintains the skin barrier by mediating differentiation, lipid metabolism and tight junction of keratinocyte

doi: 10.1038/s41598-025-01960-w

Figure Lengend Snippet: Norathyriol and PPARα shRNA lentiviral restricted promotion effects of PUR-P on keratinocyte differentiation, lipid metabolism and tight junctions. ( A – D ) Western blot and qRT-PCR assays characterized the effects of Norathyriol (PPARα antagonist) and PPARα shRNA lentivirus on the expression of differentiation-associated (K1 and K10; A ), sebum-synthesis-associated (CERS3 and FAS; B ), cornified envelope-associated (FLG, INV, and LOR; C ) and tight junction-associated (ZO-1; D ) proteins and mRNA. The original gel blot is shown in the Supplementary Information file. DRPs: keratinocyte differentiation-related proteins; SSRPs: sebum synthesis-related proteins; CEPs: cornified envelope-related proteins; TJRPs: tight junction-related proteins. N = 3; One-way ANOVA test with Tukey’s multiple comparisons test or Kruskal–Wallis test with Dunn’s multiple comparisons test; Statistical significance was defined as P < 0.05.

Article Snippet: The KD-NC and KD-PPARα groups were infected with sh-NC and sh-PPARα_1 lentivirus using the HitransG P (REVG005, Genechem, China) for 48 h. The Norathyriol group were treated with 100 μM Norathyriol, a PPARα antagonist (HY-N1029, MCE, USA) for 24 h. Subsequently, the PUR-P, Norathyriol, KD-NC, and KD-PPARα groups were treated with 10 mg/ml PUR-P for 48 and 72 h. The Solvent group were treated with equal volumes of sterile water.

Techniques: shRNA, Western Blot, Quantitative RT-PCR, Expressing

Journal: Scientific Reports

Article Title: Polysaccharide from Prinsepia utilis Royle maintains the skin barrier by mediating differentiation, lipid metabolism and tight junction of keratinocyte

doi: 10.1038/s41598-025-01960-w

Figure Lengend Snippet: PUR-P alleviates sodium lauryl sulfate (SLS)-induced skin barrier damage in the FulKutis model. Fulkutis is a two-layer skin model constructed with fibroblasts and keratinocytes as seed cells. The FulKutis model was used to evaluate the effect of PUR-P on SLS-induced skin damage. ( A ) Immunofluorescence staining was used to detect and localize the expression of a negative marker (K15) and a positive marker (Vimentin) in human-derived fibroblasts. Scale: 50 μm. ( B ) HE staining was used to observe the structural integrity of the FulKutis model in each group. Representative images of FulKutis model from CTRL, SLS, PUP-R, Norathyriol, KD-NC and KD-PPARα groups. Scale: 50 μm. C-D: TEER ( C ) and FITC-Dextran permeability ( D ) of FulKutis model in each group were measured using RE1600 epithelial cell voltage-resistance meter and Multiskan SkyHigh microplate reader, respectively, to evaluate the therapeutic effect of PUR-P on SLS-induced skin barrier damage. Ep, De, SC, SG, SS and SB indicate epidermis, dermis, stratum corneum, stratum granulosum, stratum spinosum, and stratum basale, respectively. N = 3; One-way ANOVA test with Tukey’s multiple comparisons test; Statistical significance was defined as P < 0.05.

Article Snippet: The KD-NC and KD-PPARα groups were infected with sh-NC and sh-PPARα_1 lentivirus using the HitransG P (REVG005, Genechem, China) for 48 h. The Norathyriol group were treated with 100 μM Norathyriol, a PPARα antagonist (HY-N1029, MCE, USA) for 24 h. Subsequently, the PUR-P, Norathyriol, KD-NC, and KD-PPARα groups were treated with 10 mg/ml PUR-P for 48 and 72 h. The Solvent group were treated with equal volumes of sterile water.

Techniques: Construct, Immunofluorescence, Staining, Expressing, Marker, Derivative Assay, Permeability

Journal: Scientific Reports

Article Title: Polysaccharide from Prinsepia utilis Royle maintains the skin barrier by mediating differentiation, lipid metabolism and tight junction of keratinocyte

doi: 10.1038/s41598-025-01960-w

Figure Lengend Snippet: PUR-P facilitates the expression of proteins related to differentiation, lipid metabolism and tight junctions by regulating PPARα in the FulKutis model. ( A , B ) The expression of differentiation-associated (K1 and K10; A ), sebum-synthesis-associated (CERS3 and FAS; B ), cornified envelope-associated (FLG, INV, and LOR; C ), and tight junction-associated (ZO-1; D ) proteins were identified using western blot assay in the FulKutis model. The original gel blot is shown in the Supplementary Information file. DRPs: keratinocyte differentiation-related proteins; SSRPs: sebum synthesis-related proteins; CEPs: cornified envelope-related proteins; TJRPs: tight junction-related proteins. N = 3; One-way ANOVA test with Tukey’s multiple comparisons test; Statistical significance was defined as P < 0.05.

Article Snippet: The KD-NC and KD-PPARα groups were infected with sh-NC and sh-PPARα_1 lentivirus using the HitransG P (REVG005, Genechem, China) for 48 h. The Norathyriol group were treated with 100 μM Norathyriol, a PPARα antagonist (HY-N1029, MCE, USA) for 24 h. Subsequently, the PUR-P, Norathyriol, KD-NC, and KD-PPARα groups were treated with 10 mg/ml PUR-P for 48 and 72 h. The Solvent group were treated with equal volumes of sterile water.

Techniques: Expressing, Western Blot

Journal: Scientific Reports

Article Title: Polysaccharide from Prinsepia utilis Royle maintains the skin barrier by mediating differentiation, lipid metabolism and tight junction of keratinocyte

doi: 10.1038/s41598-025-01960-w

Figure Lengend Snippet: Molecular mechanisms of PUR-P for maintaining the skin barrier. PUR-P upregulates differentiation- related (K1 and K10), proliferation- related (K16 and K17), sebum synthesis- related (CERS3, FAS, and HMGCS2), tight junction- related (ZO-1, Occludin, and Claudin-1), and cornified envelope-related (FLG, INV, and LOR) proteins in keratinocytes through activation of the PPARα pathway, thereby enhancing the skin barrier. It was obtained from Freepik ( https://www.freepik.com/free-vector/anatomical-structure-human-skin-layers-with-cross-section-labelled-parts-realistic-infographics-vector-illustration_26762467.htm#fromView=search&page=1&position=1&uuid=dc21ff73-da36-48c9-aeee-3ddafbc95021&query=skin+structure ). According to Freepik’s Terms of use, the way we use it does not re-quire permission file. For more information, please see https://www.freepik.com/legal/terms-of-use .

Article Snippet: The KD-NC and KD-PPARα groups were infected with sh-NC and sh-PPARα_1 lentivirus using the HitransG P (REVG005, Genechem, China) for 48 h. The Norathyriol group were treated with 100 μM Norathyriol, a PPARα antagonist (HY-N1029, MCE, USA) for 24 h. Subsequently, the PUR-P, Norathyriol, KD-NC, and KD-PPARα groups were treated with 10 mg/ml PUR-P for 48 and 72 h. The Solvent group were treated with equal volumes of sterile water.

Techniques: Activation Assay, Plasmid Preparation