Journal: Biological Research

Article Title: Chinese herbal formula Huayu-Qiangshen-Tongbi decoction ameliorates rheumatoid arthritis through enhancing the release of exosomal miR-125b-5p derived from adipose-derived stem cells by CD63

doi: 10.1186/s40659-025-00628-z

Figure Lengend Snippet: HQT improves LPS-reduced miR-125b-5p level in FLSs by ADSC-derived exosomes. A miR-125b-5p level detected in control, LPS-treated, LPS-treated plus co-cultured with ADSCs incubated with blank serum, 5%/15%/25% HQT-containing serum or GW4869 MH7A cells. B The size of exosomes derived from ADSC treated with or without blank serum and 25% HQT-containing serum identified by nanoparticle tracking analysis. C WB analysis of exosomal protein markers CD63 and CD81 in exosomes derived from ADSC treated with or without blank serum and 25% HQT-containing serum. D Representative images of MH7A cells co-cultured with PKH26-stained ADSC-derived exosomes. E miR-125b-5p level detected in MH7A cells treated with or without LPS plus exosomes derived from ADSC treated with blank serum or 25% HQT-containing serum. HQT: Huayu-Qiangshen-Tongbi decoction; Exo: exosomes. * P < 0.05, *** P < 0.001, **** P < 0.0001

Article Snippet: The primary antibodies used in this study were listed as follow: CD9 antibody (1:1000, #bs-5660R, Bioss), CD81 antibody (1:1000, #bs-2489R, Bioss), CD63 antibody (1:1000, #bs-1523R, Bioss), NF-κB p65 antibody (1:1000, #bs-5660R, Bioss) and GAPDH antibody (1:10000, #KC-5G5, Aksomics, Shanghai, China).

Techniques: Derivative Assay, Control, Cell Culture, Incubation, Staining

Journal: Biological Research

Article Title: Chinese herbal formula Huayu-Qiangshen-Tongbi decoction ameliorates rheumatoid arthritis through enhancing the release of exosomal miR-125b-5p derived from adipose-derived stem cells by CD63

doi: 10.1186/s40659-025-00628-z

Figure Lengend Snippet: HQT enhances the release of exosomal miR-125b-5p derived from ADSCs by CD63. A miR-125b-5p level detected in exosomes derived from control, blank serum-treated and 25% HQT-containing serum-treated ADSCs. B CD63 protein level detected in control and 25% HQT-containing serum-treated ADSCs. C CD63 protein level detected in 25% HQT-containing serum-treated ADSCs transfected with shRNA NC or CD63 shRNA. D miR-125b-5p level detected in exosomes derived from 25% HQT-containing serum-treated ADSCs transfected with or without CD63 shRNA. HQT: Huayu-Qiangshen-Tongbi decoction; NC: negative control; si: silence. * P < 0.05, ** P < 0.01, **** P < 0.0001

Article Snippet: The primary antibodies used in this study were listed as follow: CD9 antibody (1:1000, #bs-5660R, Bioss), CD81 antibody (1:1000, #bs-2489R, Bioss), CD63 antibody (1:1000, #bs-1523R, Bioss), NF-κB p65 antibody (1:1000, #bs-5660R, Bioss) and GAPDH antibody (1:10000, #KC-5G5, Aksomics, Shanghai, China).

Techniques: Derivative Assay, Control, Transfection, shRNA, Negative Control

Journal: Biological Research

Article Title: Chinese herbal formula Huayu-Qiangshen-Tongbi decoction ameliorates rheumatoid arthritis through enhancing the release of exosomal miR-125b-5p derived from adipose-derived stem cells by CD63

doi: 10.1186/s40659-025-00628-z

Figure Lengend Snippet: CD63 is the target of HQT. Network pharmacology-based analysis indicated that CD63 should be the target of Radix Dipsaci Asperoidis (Chuanxuduan) ( A ) and Glycyrrhizae Radix et Rhizoma (Gancao) ( B )

Article Snippet: The primary antibodies used in this study were listed as follow: CD9 antibody (1:1000, #bs-5660R, Bioss), CD81 antibody (1:1000, #bs-2489R, Bioss), CD63 antibody (1:1000, #bs-1523R, Bioss), NF-κB p65 antibody (1:1000, #bs-5660R, Bioss) and GAPDH antibody (1:10000, #KC-5G5, Aksomics, Shanghai, China).

Techniques:

Journal: Biological Research

Article Title: Chinese herbal formula Huayu-Qiangshen-Tongbi decoction ameliorates rheumatoid arthritis through enhancing the release of exosomal miR-125b-5p derived from adipose-derived stem cells by CD63

doi: 10.1186/s40659-025-00628-z

Figure Lengend Snippet: HQT suppresses CD63 degradation in ADSCs. A Transcription activity of CD63 gene was analyzed by relative luciferase reporter activity assay in control and 25% HQT-containing serum-treated ADSCs. B CD63 protein level detected in control, blank serum-treated and 25% HQT-containing serum-treated ADSCs after CHX treatment. HQT: Huayu-Qiangshen-Tongbi decoction

Article Snippet: The primary antibodies used in this study were listed as follow: CD9 antibody (1:1000, #bs-5660R, Bioss), CD81 antibody (1:1000, #bs-2489R, Bioss), CD63 antibody (1:1000, #bs-1523R, Bioss), NF-κB p65 antibody (1:1000, #bs-5660R, Bioss) and GAPDH antibody (1:10000, #KC-5G5, Aksomics, Shanghai, China).

Techniques: Activity Assay, Luciferase, Control

Journal: Biological Research

Article Title: Chinese herbal formula Huayu-Qiangshen-Tongbi decoction ameliorates rheumatoid arthritis through enhancing the release of exosomal miR-125b-5p derived from adipose-derived stem cells by CD63

doi: 10.1186/s40659-025-00628-z

Figure Lengend Snippet: Schematic diagram of molecular mechanisms for the current study. HQT suppressed CD63 degradation to facilitate the release of exosomal miR-125b-5p derived from ADSCs and subsequently exosomal miR-125b-5p was transferred into FLSs to attenuate RA-induced inflammation

Article Snippet: The primary antibodies used in this study were listed as follow: CD9 antibody (1:1000, #bs-5660R, Bioss), CD81 antibody (1:1000, #bs-2489R, Bioss), CD63 antibody (1:1000, #bs-1523R, Bioss), NF-κB p65 antibody (1:1000, #bs-5660R, Bioss) and GAPDH antibody (1:10000, #KC-5G5, Aksomics, Shanghai, China).

Techniques: Derivative Assay

Journal: Cancer Medicine

Article Title: Exosome‐mediated delivery of miR‐204‐5p inhibits tumor growth and chemoresistance

doi: 10.1002/cam4.3248

Figure Lengend Snippet: Characteristics of exosomes derived from miR‐204‐5p‐overexpressing HEK293T cells. A, The markers of exosomes (CD63 and Flotillin‐2) were detected in HEK293T cells and exosomes by Western blot. B, The transmission electron micrograph showed roundshaped vesicles with bilayered membranes ranging from 100 nm to 150 nm in diameter released by HEK293T cells. Scale bar = 200 nm. C, 293T EXOs size distribution was measured by Zetasizer. D, Real‐time qRT‐PCR revealed that the level of miR‐204‐5p was higher in 293T‐204 cells and miR‐204 EXO than miR‐204‐3p. *** P < .001. Shown are mean ± SEM from three independent experiments

Article Snippet: The protein samples of cells and exosomes were detected with antibodies against CD63 (1:1000; BOSTER), Flotillin‐2 (1:500; Santa Cruz), RAB22A (1:1000; Proteintech), Bcl2 (1:1000; Santa Cruz) and β‐actin (1:2000; Thermo) as we previously described.,

Techniques: Derivative Assay, Western Blot, Transmission Assay, Quantitative RT-PCR