Mechanism of Bazi Bushen capsule in delaying the senescence of mesenchymal stem cells based on network pharmacology and experimental validation
Yaping Zhang,
Tongxing Wang,
Yanfei Song,
Meng Chen,
Bin Hou,
Bing Yao,
Kun Ma,
Yahui Song,
Siwei Wang,
Dan Zhang,
Junqing Liang,
Cong Wei
Affiliations
Yaping Zhang
Graduate School, Hebei University of Chinese Medicine, Shijiazhuang, 050091, China
Tongxing Wang
National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China; Key Laboratory of State Administration of TCM (Cardio-Cerebral Vessel Collateral Disease), Shijiazhuang, 050035, China
Yanfei Song
National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China; Shijiazhuang Compound Traditional Chinese Medicine Technology Innovation Center, Shijiazhuang, 050035, China
Meng Chen
National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China; Shijiazhuang Compound Traditional Chinese Medicine Technology Innovation Center, Shijiazhuang, 050035, China
Bin Hou
National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China
Bing Yao
National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China; Shijiazhuang Compound Traditional Chinese Medicine Technology Innovation Center, Shijiazhuang, 050035, China
Kun Ma
National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China; Hebei Clinical Research Center of Cardiovascular Disease of Traditional Chinese Medicine, Shijiazhuang, 050035, China
Yahui Song
National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China
Siwei Wang
Graduate School, Hebei University of Chinese Medicine, Shijiazhuang, 050091, China; National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China
Dan Zhang
Graduate School, Hebei University of Chinese Medicine, Shijiazhuang, 050091, China; National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China
Junqing Liang
National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China; Corresponding author. National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China.
Cong Wei
Graduate School, Hebei University of Chinese Medicine, Shijiazhuang, 050091, China; National Key Laboratory for Innovation and Transformation of Luobing Theory, Shijiazhuang, 050035, China; High-level TCM Key Disciplines of National Administration of Traditional Chinese Medicine—Luobing Theory, Hebei Province, Shijiazhuang, 050035, China; Corresponding author. Graduate School, Hebei University of Chinese Medicine, Shijiazhuang, 050091, China.
Ageing is becoming an increasingly serious problem; therefore, there is an urgent need to find safe and effective anti-ageing drugs. Aims: To investigate the effects of Bazi Bushen capsule (BZBS) on the senescence of mesenchymal stem cells (MSCs) and explore its mechanism of action. Methods: Network pharmacology was used to predict the targets of BZBS in delaying senescence in MSCs. For in vitro studies, MSCs were treated with D-gal, BZBS, and NMN, and cell viability, cell senescence, stemness-related genes, and cell cycle were studied using cell counting kit-8 (CCK-8) assay, SA-β-galactosidase (SA-β-gal) staining, Quantitative Real-Time PCR (qPCR) and flow cytometry (FCM), respectively. Alkaline phosphatase (ALP), alizarin red, and oil red staining were used to determine the osteogenic and lipid differentiation abilities of MSCs. Finally, the expression of senescence-related genes and cyclin-related factors was detected by qPCR and western blotting. Results: Network pharmacological analysis suggested that BZBS delayed cell senescence by interfering in the cell cycle. Our in vitro studies suggested that BZBS could significantly increase cell viability (P < 0.01), decrease the quantity of β-galactosidase+ cells (P < 0.01), downregulate p16 and p21 (P < 0.05, P < 0.01), improve adipogenic and osteogenic differentiation, and upregulate Nanog, OCT4 and SOX2 genes (P < 0.05, P < 0.01) in senescent MSCs. Moreover, BZBS significantly reduced the proportion of senescent MSCs in the G0/G1 phase (P < 0.01) and enhanced the expression of CDK4, Cyclin D1, and E2F1 (P < 0.05, P < 0.01, respectively). Upon treatment with HY-50767A, a CDK4 inhibitor, the upregulation of E2F1 was no longer observed in the BZBS group. Conclusions: BZBS can protect MSCs against D-gal-induced senescence, which may be associated with cell cycle regulation via the Cyclin D1/CDK4/E2F1 signalling pathway.
