Journal: Journal of Cellular and Molecular Medicine

Article Title: Casticin From Sanse Powder‐ In Vitro Release Medium Inhibits FAK / PI3K / AKT Phosphorylation Reducing Synovial Senescence and Alleviates Inflammation in Knee Osteoarthritis

doi: 10.1111/jcmm.71391

Figure Lengend Snippet: SSP‐IVRM alleviates synovial inflammation and pathological progression in KOA rats. (A) ELISA was used to detect the levels of IL‐1β, IL‐6, CXCL1 and CXCL3 in the serum of rats in each group; (B) Representative images of HE, Masson and Sirius Red staining of knee synovial tissues from each group, along with Krenn's scoring; (C) Representative images of IL‐1β immunofluorescence in knee synovial tissues from each group; (D) Western blot analysis of the protein expression levels of MMP3, MMP9, COL I and TGF‐β in the synovial tissues of rats in each group. (E) qRT‐PCR analysis of MMP3, MMP9, COL1A1 and TGF‐β mRNA expression in synovial tissues. Statistical graphs are presented as Mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 versus Ctrl group; # p < 0.05, ## p < 0.01, ### p < 0.001 versus KOA group.

Article Snippet: DMEM high‐glucose medium (Cat# C11995500CP) and fetal bovine serum (Cat# 10099‐141) were purchased from GIBCO (USA); LPS (Cat# L2880) was obtained from Sigma‐Aldrich (USA); Casticin (Cat# B20835 , ≥ 98%) was purchased from Yuanye Biotechnology (China); MIR002 (Cat# HY‐143412), Y15 (Cat# HY‐12444) and VSLRGDTRG acetate (Cat# HY‐P10941A) were obtained from MCE (China); Senescence‐Tracker Senescent Cell Fluorescent Probe (Cat# C0603), Enhanced Mitochondrial Membrane Potential Assay Kit (Cat# C2003S), Reactive Oxygen Species Assay Kit (Cat# S0035S) and Cell Counting Kit‐8 (Cat# C0038) were purchased from Beyotime Biotechnology (China); Rat CXCL1 ELISA Kit (Cat# E‐EL‐R0003), Rat CXCL3 ELISA Kit (Cat# E‐EL‐R0697), Rat IL‐6 ELISA Kit (Cat# E‐EL‐R0015) and Rat IL‐1β ELISA Kit (Cat# E‐EL‐R0012) were obtained from Elabscience (China); TGF‐β Antibody (Cat# 3711), CDC2 Antibody (Cat# 77055), FAK Antibody (Cat# 3285), PI3 Kinase p85 Antibody (Cat# 4292), AKT Antibody (Cat# 9272) and Phospho‐FAK (Tyr397) Antibody (Cat# 3283) were purchased from CST (USA); Phospho‐PI3K p85 (Tyr458)/p55 (Tyr199) Antibody (Cat# AF3242) was obtained from Affinity (China); Phospho‐AKT (Ser473) Antibody (Cat# 80455‐1‐RR), IL‐1β Antibody (Cat# 26048‐1‐AP), IL‐6 Antibody (Cat# 21865‐1‐AP), MMP3 Antibody (Cat# 17873‐1‐AP), MMP9 Antibody (Cat# 10375‐2‐AP), p53 Antibody (Cat# 10442‐1‐AP), p21 Antibody (Cat# 10355‐1‐AP) and GAPDH Antibody (Cat# 60004‐1‐Ig) were purchased from Proteintech (China).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Immunofluorescence, Western Blot, Analysis, Expressing, Quantitative RT-PCR

Journal: Journal of Cellular and Molecular Medicine

Article Title: Casticin From Sanse Powder‐ In Vitro Release Medium Inhibits FAK / PI3K / AKT Phosphorylation Reducing Synovial Senescence and Alleviates Inflammation in Knee Osteoarthritis

doi: 10.1111/jcmm.71391

Figure Lengend Snippet: CAS alleviates synovial inflammation and cellular senescence in KOA rats. (A) Representative images of HE, Masson and Sirius Red staining of synovial tissue from rat knee joints in each group, along with Krenn's scores; (B) Serum levels of IL‐1β, IL‐6, CXCL1 and CXCL3 in each group of rats as measured by ELISA; (C) Representative immunofluorescence images of IL‐1β in synovial tissue from rat knee joints in each group; (D) Protein expression levels of MMP3, MMP9, COL I, TGF‐β, p53, p21, CDC2 and CCNA1 in synovial tissue from each rat group as determined by Western Blot. (E) qRT‐PCR analysis of the mRNA expression levels of MMP3, MMP9, COL1A1, TGF‐β, p53, p21/CDKN1A, CDC2 and CCNA1 in synovial tissues from each rat group. Statistical graphs are presented as Mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 versus Ctrl group; # p < 0.05, ## p < 0.01, ### p < 0.001 versus KOA group.

Article Snippet: DMEM high‐glucose medium (Cat# C11995500CP) and fetal bovine serum (Cat# 10099‐141) were purchased from GIBCO (USA); LPS (Cat# L2880) was obtained from Sigma‐Aldrich (USA); Casticin (Cat# B20835 , ≥ 98%) was purchased from Yuanye Biotechnology (China); MIR002 (Cat# HY‐143412), Y15 (Cat# HY‐12444) and VSLRGDTRG acetate (Cat# HY‐P10941A) were obtained from MCE (China); Senescence‐Tracker Senescent Cell Fluorescent Probe (Cat# C0603), Enhanced Mitochondrial Membrane Potential Assay Kit (Cat# C2003S), Reactive Oxygen Species Assay Kit (Cat# S0035S) and Cell Counting Kit‐8 (Cat# C0038) were purchased from Beyotime Biotechnology (China); Rat CXCL1 ELISA Kit (Cat# E‐EL‐R0003), Rat CXCL3 ELISA Kit (Cat# E‐EL‐R0697), Rat IL‐6 ELISA Kit (Cat# E‐EL‐R0015) and Rat IL‐1β ELISA Kit (Cat# E‐EL‐R0012) were obtained from Elabscience (China); TGF‐β Antibody (Cat# 3711), CDC2 Antibody (Cat# 77055), FAK Antibody (Cat# 3285), PI3 Kinase p85 Antibody (Cat# 4292), AKT Antibody (Cat# 9272) and Phospho‐FAK (Tyr397) Antibody (Cat# 3283) were purchased from CST (USA); Phospho‐PI3K p85 (Tyr458)/p55 (Tyr199) Antibody (Cat# AF3242) was obtained from Affinity (China); Phospho‐AKT (Ser473) Antibody (Cat# 80455‐1‐RR), IL‐1β Antibody (Cat# 26048‐1‐AP), IL‐6 Antibody (Cat# 21865‐1‐AP), MMP3 Antibody (Cat# 17873‐1‐AP), MMP9 Antibody (Cat# 10375‐2‐AP), p53 Antibody (Cat# 10442‐1‐AP), p21 Antibody (Cat# 10355‐1‐AP) and GAPDH Antibody (Cat# 60004‐1‐Ig) were purchased from Proteintech (China).

Techniques: Staining, Tissue, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Expressing, Western Blot, Quantitative RT-PCR, Analysis

Journal: Journal of Cellular and Molecular Medicine

Article Title: Casticin From Sanse Powder‐ In Vitro Release Medium Inhibits FAK / PI3K / AKT Phosphorylation Reducing Synovial Senescence and Alleviates Inflammation in Knee Osteoarthritis

doi: 10.1111/jcmm.71391

Figure Lengend Snippet: CAS improves inflammation and cellular senescence in LPS‐induced rat synovial fibroblast‐like cells. (A) CCK‐8 assay to evaluate the cytotoxic effects of CAS on FLS; (B) ELISA to measure the levels of IL‐1β, IL‐6, CXCL1 and CXCL3 in the culture media of rat FLS from each group; (C) Representative immunofluorescence images of IL‐1β and IL‐6 in FLS from each group; (D) Western blot analysis of protein expression levels of MMP3, MMP9, COL I, TGF‐β, p53, p21, CDC2 and CCNA1 in FLS from each group; (E) CM‐H2DCFDA probe to assess ROS levels in FLS from each group; (F) JC‐1 probe to detect mitochondrial membrane potential levels in FLS from each group; (G) Representative images of senescent cells labelled by Senescence‐Tracker probe in FLS from each group. Statistical graphs are presented as Mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 versus Ctrl group; # p < 0.05, ## p < 0.01, ### p < 0.001 versus LPS group.

Article Snippet: DMEM high‐glucose medium (Cat# C11995500CP) and fetal bovine serum (Cat# 10099‐141) were purchased from GIBCO (USA); LPS (Cat# L2880) was obtained from Sigma‐Aldrich (USA); Casticin (Cat# B20835 , ≥ 98%) was purchased from Yuanye Biotechnology (China); MIR002 (Cat# HY‐143412), Y15 (Cat# HY‐12444) and VSLRGDTRG acetate (Cat# HY‐P10941A) were obtained from MCE (China); Senescence‐Tracker Senescent Cell Fluorescent Probe (Cat# C0603), Enhanced Mitochondrial Membrane Potential Assay Kit (Cat# C2003S), Reactive Oxygen Species Assay Kit (Cat# S0035S) and Cell Counting Kit‐8 (Cat# C0038) were purchased from Beyotime Biotechnology (China); Rat CXCL1 ELISA Kit (Cat# E‐EL‐R0003), Rat CXCL3 ELISA Kit (Cat# E‐EL‐R0697), Rat IL‐6 ELISA Kit (Cat# E‐EL‐R0015) and Rat IL‐1β ELISA Kit (Cat# E‐EL‐R0012) were obtained from Elabscience (China); TGF‐β Antibody (Cat# 3711), CDC2 Antibody (Cat# 77055), FAK Antibody (Cat# 3285), PI3 Kinase p85 Antibody (Cat# 4292), AKT Antibody (Cat# 9272) and Phospho‐FAK (Tyr397) Antibody (Cat# 3283) were purchased from CST (USA); Phospho‐PI3K p85 (Tyr458)/p55 (Tyr199) Antibody (Cat# AF3242) was obtained from Affinity (China); Phospho‐AKT (Ser473) Antibody (Cat# 80455‐1‐RR), IL‐1β Antibody (Cat# 26048‐1‐AP), IL‐6 Antibody (Cat# 21865‐1‐AP), MMP3 Antibody (Cat# 17873‐1‐AP), MMP9 Antibody (Cat# 10375‐2‐AP), p53 Antibody (Cat# 10442‐1‐AP), p21 Antibody (Cat# 10355‐1‐AP) and GAPDH Antibody (Cat# 60004‐1‐Ig) were purchased from Proteintech (China).

Techniques: CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Western Blot, Analysis, Expressing, Membrane

Journal: Journal of Cellular and Molecular Medicine

Article Title: Casticin From Sanse Powder‐ In Vitro Release Medium Inhibits FAK / PI3K / AKT Phosphorylation Reducing Synovial Senescence and Alleviates Inflammation in Knee Osteoarthritis

doi: 10.1111/jcmm.71391

Figure Lengend Snippet: CAS reduces the inflammatory state of rat synovial fibroblast‐like cells by lowering cellular senescence levels. (A) Representative images of senescent positive cells in rat FLSs from each group detected using the Senescence‐Tracker probe; (B) Mitochondrial membrane potential levels in rat FLSs from each group measured by the JC‐1 probe; (C) Intracellular ROS levels in rat FLSs from each group assessed with the CM‐H2DCFDA probe; (D) Protein expression levels of MMP3, MMP9, COL I, TGF‐β, p53, p21, CDC2 and CCNA1 in rat FLSs from each group detected by Western blotting (WB); (E) Representative immunofluorescence images of IL‐1β and IL‐6 in rat FLSs from each group; (F) Levels of IL‐1β, IL‐6, CXCL1 and CXCL3 in the culture supernatant of rat FLSs from each group measured by ELISA. Statistical graphs are presented as Mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 versus Ctrl group; # p < 0.05, ## p < 0.01, ### p < 0.001 versus LPS group; △ p < 0.05, △△ p < 0.01, △△△ p < 0.001 versus MIR002 group.

Article Snippet: DMEM high‐glucose medium (Cat# C11995500CP) and fetal bovine serum (Cat# 10099‐141) were purchased from GIBCO (USA); LPS (Cat# L2880) was obtained from Sigma‐Aldrich (USA); Casticin (Cat# B20835 , ≥ 98%) was purchased from Yuanye Biotechnology (China); MIR002 (Cat# HY‐143412), Y15 (Cat# HY‐12444) and VSLRGDTRG acetate (Cat# HY‐P10941A) were obtained from MCE (China); Senescence‐Tracker Senescent Cell Fluorescent Probe (Cat# C0603), Enhanced Mitochondrial Membrane Potential Assay Kit (Cat# C2003S), Reactive Oxygen Species Assay Kit (Cat# S0035S) and Cell Counting Kit‐8 (Cat# C0038) were purchased from Beyotime Biotechnology (China); Rat CXCL1 ELISA Kit (Cat# E‐EL‐R0003), Rat CXCL3 ELISA Kit (Cat# E‐EL‐R0697), Rat IL‐6 ELISA Kit (Cat# E‐EL‐R0015) and Rat IL‐1β ELISA Kit (Cat# E‐EL‐R0012) were obtained from Elabscience (China); TGF‐β Antibody (Cat# 3711), CDC2 Antibody (Cat# 77055), FAK Antibody (Cat# 3285), PI3 Kinase p85 Antibody (Cat# 4292), AKT Antibody (Cat# 9272) and Phospho‐FAK (Tyr397) Antibody (Cat# 3283) were purchased from CST (USA); Phospho‐PI3K p85 (Tyr458)/p55 (Tyr199) Antibody (Cat# AF3242) was obtained from Affinity (China); Phospho‐AKT (Ser473) Antibody (Cat# 80455‐1‐RR), IL‐1β Antibody (Cat# 26048‐1‐AP), IL‐6 Antibody (Cat# 21865‐1‐AP), MMP3 Antibody (Cat# 17873‐1‐AP), MMP9 Antibody (Cat# 10375‐2‐AP), p53 Antibody (Cat# 10442‐1‐AP), p21 Antibody (Cat# 10355‐1‐AP) and GAPDH Antibody (Cat# 60004‐1‐Ig) were purchased from Proteintech (China).

Techniques: Membrane, Expressing, Western Blot, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Casticin From Sanse Powder‐ In Vitro Release Medium Inhibits FAK / PI3K / AKT Phosphorylation Reducing Synovial Senescence and Alleviates Inflammation in Knee Osteoarthritis

doi: 10.1111/jcmm.71391

Figure Lengend Snippet: CAS inhibits FAK/PI3K/AKT phosphorylation process to reduce synovial cell senescence and improve synovial inflammation. (A) Representative immunofluorescence images of p‐FAK and p‐PI3K in FLS from each rat group; (B) Western blot analysis of protein expression levels of p‐FAK, FAK, p‐PI3K, PI3K, p‐AKT and AKT in FLS from each group; (C) Western blot analysis of protein expression levels of MMP9, COL I, TGF‐β, p53, p21, CDC2 and CCNA1 in FLS from each group; (D) Assessment of mitochondrial membrane potential in FLS from each group using the JC‐1 probe; (E) Detection of ROS levels in FLS from each group using the CM‐H2DCFDA probe; (F) Representative images of senescent FLS detected with the Senescence‐Tracker probe in each group; (G) ELISA analysis of IL‐1β, IL‐6, CXCL1 and CXCL3 levels in the culture medium of FLS from each group. Statistical graphs are presented as Mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 versus Ctrl group; # p < 0.05, ## p < 0.01, ### p < 0.001 versus LPS group; △ p < 0.05, △△ p < 0.01, △△△ p < 0.001 versus CAS group.

Article Snippet: DMEM high‐glucose medium (Cat# C11995500CP) and fetal bovine serum (Cat# 10099‐141) were purchased from GIBCO (USA); LPS (Cat# L2880) was obtained from Sigma‐Aldrich (USA); Casticin (Cat# B20835 , ≥ 98%) was purchased from Yuanye Biotechnology (China); MIR002 (Cat# HY‐143412), Y15 (Cat# HY‐12444) and VSLRGDTRG acetate (Cat# HY‐P10941A) were obtained from MCE (China); Senescence‐Tracker Senescent Cell Fluorescent Probe (Cat# C0603), Enhanced Mitochondrial Membrane Potential Assay Kit (Cat# C2003S), Reactive Oxygen Species Assay Kit (Cat# S0035S) and Cell Counting Kit‐8 (Cat# C0038) were purchased from Beyotime Biotechnology (China); Rat CXCL1 ELISA Kit (Cat# E‐EL‐R0003), Rat CXCL3 ELISA Kit (Cat# E‐EL‐R0697), Rat IL‐6 ELISA Kit (Cat# E‐EL‐R0015) and Rat IL‐1β ELISA Kit (Cat# E‐EL‐R0012) were obtained from Elabscience (China); TGF‐β Antibody (Cat# 3711), CDC2 Antibody (Cat# 77055), FAK Antibody (Cat# 3285), PI3 Kinase p85 Antibody (Cat# 4292), AKT Antibody (Cat# 9272) and Phospho‐FAK (Tyr397) Antibody (Cat# 3283) were purchased from CST (USA); Phospho‐PI3K p85 (Tyr458)/p55 (Tyr199) Antibody (Cat# AF3242) was obtained from Affinity (China); Phospho‐AKT (Ser473) Antibody (Cat# 80455‐1‐RR), IL‐1β Antibody (Cat# 26048‐1‐AP), IL‐6 Antibody (Cat# 21865‐1‐AP), MMP3 Antibody (Cat# 17873‐1‐AP), MMP9 Antibody (Cat# 10375‐2‐AP), p53 Antibody (Cat# 10442‐1‐AP), p21 Antibody (Cat# 10355‐1‐AP) and GAPDH Antibody (Cat# 60004‐1‐Ig) were purchased from Proteintech (China).

Techniques: Phospho-proteomics, Immunofluorescence, Western Blot, Analysis, Expressing, Membrane, Enzyme-linked Immunosorbent Assay

Journal: Molecular Neurobiology

Article Title: Silibinin Alleviates Cognitive Dysfunction, Biochemical Alterations, and Hippocampal Structural Changes Induced by 5 days of Sleep Deprivation with or Without a Subsequent 24-H Recovery in Rats

doi: 10.1007/s12035-026-06195-y

Figure Lengend Snippet: Effect of silibinin administration on A BDNF (ng/ml) and B CaMKII (ng/mg) in sleep-deprived rats with and without consequent 24-h recovery. Data are represented as mean ± SD ( n = 8); group comparisons were performed using one-way ANOVA, followed by Tukey’s multiple-comparisons test. * P < 0.05, ** P < 0.01, and *** P < 0.001 versus the control group; $ P < 0.05, $$ P < 0.01, and $$$ P < 0.001 versus the SD group; # P < 0.05, ## P < 0.01, and ### P < 0.001 versus the SD + R group. SD: sleep-deprived untreated group; SD + R: sleep-deprived untreated group with recovery; SD + Sil: sleep-deprived group treated with silibinin; SD + Sil + R: sleep-deprived group treated with silibinin followed by recovery. BDNF: brain‑derived neurotrophic factor; CaMKII: calcium/calmodulin‑dependent protein kinase II

Article Snippet: Furthermore, hippocampal tissue homogenates were prepared to determine BDNF (Boster Biological Technology, Pleasanton, CA, USA; Cat. No. EK0308) and CaMKII (BlueGene Biotech, Shanghai, China; Cat. No. E02C0682) using commercially available ELISA kits supplied by Bio-KITS (Cairo, Egypt), according to the manufacturers' instructions.

Techniques: Control