PMID- 36804262
OWN - NLM
STAT- MEDLINE
DCOM- 20230807
LR  - 20230919
IS  - 2542-5641 (Electronic)
IS  - 0366-6999 (Print)
IS  - 0366-6999 (Linking)
VI  - 136
IP  - 15
DP  - 2023 Aug 5
TI  - Baicalin attenuates dexamethasone-induced apoptosis of bone marrow mesenchymal 
      stem cells by activating the hedgehog signaling pathway.
PG  - 1839-1847
LID - 10.1097/CM9.0000000000002113 [doi]
AB  - BACKGROUND: Perturbations in bone marrow mesenchymal stem cell (BMSC) 
      differentiation play an important role in steroid-induced osteonecrosis of the 
      femoral head (SONFH). At present, studies on SONFH concentrate upon the balance 
      within BMSC osteogenic and adipogenic differentiation. However, BMSC apoptosis as 
      well as proliferation are important prerequisites in their differentiation. The 
      hedgehog (HH) signaling pathway regulates bone cell apoptosis. Baicalin (BA), a 
      well-known compound in traditional Chinese medicine, can affect the proliferation 
      and apoptosis of numerous cell types via HH signaling. However, the potential 
      role and mechanisms of BA on BMSCs are unclear. Thus, we aimed to explore the 
      role of BA in dexamethasone (Dex)-induced BMSC apoptosis in this study. METHODS: 
      Primary BMSCs were treated with 10 -6 mol/L Dex alone or with 5.0 mumol/L, 10.0 
      mumol/L, or 50.0 mumol/L BA for 24 hours followed by co-treatment with 5.0 mumol/L, 
      10.0 mumol/L, or 50.0 mumol/L BA and 10 -6 mol/L Dex. Cell viability was assayed 
      through the Cell Counting Kit-8 (CCK-8). Cell apoptosis was evaluated using 
      Annexin V-fluorescein isothiocyanate/propidium iodide (PI) staining followed by 
      flow cytometry. The imaging and counting, respectively, of Hochest 
      33342/PI-stained cells were used to assess the morphological characteristics and 
      proportion of apoptotic cells. To quantify the apoptosis-related proteins (e.g., 
      apoptosis regulator BAX [Bax], B-cell lymphoma 2 [Bcl-2], caspase-3, and cleaved 
      caspase-3) and HH signaling pathway proteins, western blotting was used. A 
      HH-signaling pathway inhibitor was used to demonstrate that BA exerts its 
      anti-apoptotic effects via the HH signaling pathway. RESULTS: The results of 
      CCK-8, Hoechst 33342/PI-staining, and flow cytometry showed that BA did not 
      significantly promote cell proliferation (CCK-8: 0 mumol/L, 100%; 2.5 mumol/L, 
      98.58%; 5.0 mumol/L, 95.18%; 10.0 mumol/L, 98.11%; 50.0 mumol/L, 99.38%, F   = 
       2.33, P   >  0.05), but it did attenuate the effect of Dex on apoptosis (Hoechst 
      33342/PI-staining: Dex+ 50.0 mumol/L BA, 12.27% vs. Dex, 39.27%, t  = 20.62; flow 
      cytometry: Dex + 50.0 mumol/L BA, 12.68% vs. Dex, 37.43%, t  = 11.56; Both P 
       < 0.05). The results of western blotting analysis showed that BA reversed 
      Dex-induced apoptosis by activating the HH signaling pathway, which 
      down-regulated the expression of Bax, cleaved-caspase 3, and suppressor of fused 
      (SUFU) while up-regulating Bcl-2, sonic hedgehog (SHH), and zinc finger protein 
      GLI-1 (GLI-1) expression (Bax/Bcl-2: Dex+ 50.0 mumol/L BA, 1.09 vs. Dex, 2.76, t 
       = 35.12; cleaved caspase-3/caspase-3: Dex + 50.0 mumol/L BA, 0.38 vs . Dex, 0.73, 
      t  = 10.62; SHH: Dex + 50.0 mumol/L BA, 0.50 vs . Dex, 0.12, t  = 34.01; SUFU: 
      Dex+ 50.0 mumol/L BA, 0.75 vs . Dex, 1.19, t  = 10.78; GLI-1: Dex+ 50.0 mumol/L BA, 
      0.40 vs . Dex, 0.11, t  = 30.68. All P  < 0.05). CONCLUSIONS: BA antagonizes 
      Dex-induced apoptosis of human BMSCs by activating the HH signaling pathway. It 
      is a potential candidate for preventing SONFH.
CI  - Copyright (c) 2023 The Chinese Medical Association, produced by Wolters Kluwer, 
      Inc. under the CC-BY-NC-ND license.
FAU - Jia, Bin
AU  - Jia B
AD  - Department of Joint Surgery, The Affiliated Hospital of Qingdao University, 
      Qingdao, Shandong 266003, China.
AD  - Medical Department, Qingdao University, Qingdao, Shandong 266071, China.
FAU - Jiang, Yaping
AU  - Jiang Y
AD  - Department of Oral Implantology, The Affiliated Hospital of Qingdao University, 
      Qingdao, Shandong 266003, China.
FAU - Yao, Yao
AU  - Yao Y
AD  - Medical Department, Qingdao University, Qingdao, Shandong 266071, China.
FAU - Xu, Yingxing
AU  - Xu Y
AD  - Department of Joint Surgery, The Affiliated Hospital of Qingdao University, 
      Qingdao, Shandong 266003, China.
AD  - Medical Department, Qingdao University, Qingdao, Shandong 266071, China.
FAU - Wang, Yingzhen
AU  - Wang Y
AD  - Department of Joint Surgery, The Affiliated Hospital of Qingdao University, 
      Qingdao, Shandong 266003, China.
FAU - Li, Tao
AU  - Li T
AD  - Department of Joint Surgery, The Affiliated Hospital of Qingdao University, 
      Qingdao, Shandong 266003, China.
LA  - eng
PT  - Journal Article
DEP - 20230805
PL  - China
TA  - Chin Med J (Engl)
JT  - Chinese medical journal
JID - 7513795
RN  - 0 (Hedgehog Proteins)
RN  - 0 (bcl-2-Associated X Protein)
RN  - EC 3.4.22.- (Caspase 3)
RN  - 347Q89U4M5 (baicalin)
RN  - 0 (Apoptosis Regulatory Proteins)
RN  - 7S5I7G3JQL (Dexamethasone)
SB  - IM
MH  - Humans
MH  - *Hedgehog Proteins/metabolism
MH  - bcl-2-Associated X Protein
MH  - Caspase 3/metabolism
MH  - Signal Transduction/physiology
MH  - Apoptosis
MH  - Apoptosis Regulatory Proteins/pharmacology
MH  - Dexamethasone/pharmacology
MH  - *Mesenchymal Stem Cells/metabolism
MH  - Bone Marrow Cells
PMC - PMC10406080
COIS- None.
EDAT- 2023/02/23 06:00
MHDA- 2023/08/07 06:41
PMCR- 2023/08/05
CRDT- 2023/02/22 08:35
PHST- 2023/08/07 06:41 [medline]
PHST- 2023/02/23 06:00 [pubmed]
PHST- 2023/02/22 08:35 [entrez]
PHST- 2023/08/05 00:00 [pmc-release]
AID - 00029330-990000000-00353 [pii]
AID - CMJ-2021-2592 [pii]
AID - 10.1097/CM9.0000000000002113 [doi]
PST - epublish
SO  - Chin Med J (Engl). 2023 Aug 5;136(15):1839-1847. doi: 
      10.1097/CM9.0000000000002113.