PMID- 35951069
OWN - NLM
STAT- MEDLINE
DCOM- 20221226
LR  - 20221226
IS  - 1078-6791 (Print)
IS  - 1078-6791 (Linking)
VI  - 29
IP  - 1
DP  - 2023 Jan
TI  - ALP Inhibitors Inhibit Inflammatory Responses and Osteoblast Differentiation in 
      hVIC via AKT-ERK Pathways.
PG  - 58-65
LID - AT7537 [pii]
AB  - OBJECTIVE: The aim of this study was to explore the calcification process of 
      aortic valve interstitial cells and its potential association with osteogenic 
      differentiation and alkaline phosphatase (ALP) activity. METHODS: The study 
      patients were divided into 3 groups: the control group, the osteogenic induction 
      medium (OM) group and the OM+ALP inhibitor group. Cell calcification was measured 
      by alizarin red S staining and alizarin red S dye released by extracellular 
      matrix (ECM) was quantified by spectrophotometry. Immunohistochemical staining 
      was performed on valve tissues of patients harboring calcified and non-calcified 
      aortic valve disease. Expression of bone morphogenetic protein (BMP), runt 
      related transcription factor 2 (RUNX2), osteocalcin and osteopontin (OPN), was 
      evaluated using immunohistochemistry and expression of osteogenic specific 
      markers (BMP, RUNX2 and OPN) was detected using Wesern blot analysis. RNA 
      sequencing was analyzed to further study the exact mechanism of ALP inhibitors in 
      terms of inhibiting the osteogenic differentiation of valvular interstitial cells 
      (VIC). The mRNA levels of tumor necrosis factor alpha (TNF-alpha), Toll-like receptor 
      4 (TLR4) and NOD-like receptor thermal protein domain associated protein 3 
      (NLRP3), were detected using reverse-transcription quantitative polymerase chain 
      reaction (RT-qPCR). In addition, Western blot analysis was performed to evaluate 
      the expression of phosphorylated extracellular regulated protein kinases (ERK), 
      nuclear factor kappa B inhibitor alpha (IkappaBalpha) and protein kinase B (AKT) in protein. 
      RESULTS: Alizarin red staining was positive in the OM and OM+ALP inhibitor 
      groups, and calcified nodules were formed in VIC, which showed a significant 
      difference compared with the control group (P < .05). The semi-quantitative level 
      of calcification in the OM group was higher than in the control group (P < .05), 
      and the semi-quantitative level of calcification in the OM+ALP inhibitor group 
      was lower than in the OM group (P < .05). ALP staining intensity, ALP activity 
      and messenger RNA (mRNA) levels of BMP, RUNX2, osteocalcin, OPN, ERK, IkappaBalpha, AKT, 
      TNF-alpha, Toll-like receptor 4 (TLR4) and NLRP3 inflammasome (NLRP3) in the OM group 
      were higher than in the control group (P < .05). ALP staining intensity, ALP 
      activity and mRNA expressions of BMP, RUNX2, osteocalcin, OPN, phosphorylated 
      ERK, IkappaBalpha, AKT, TNF-alpha and NLRP3 in the OM+ALP inhibitor group were lower than in 
      the OM group (P < .05). Compared with the control group, 723 genes were 
      upregulated and 248 genes were downregulated in the OM group. Compared with the 
      OM group, 352 genes were upregulated and 586 genes were downregulated in the 
      OM+ALP inhibitor group. CONCLUSION: We suggest that ALP inhibitors have potential 
      in terms of inhibiting the inflammatory response and osteoblast differentiation 
      of human VIC (hVIC) via the TLR4, AKT, ERK and NLRP3 pathways.
FAU - Zhang, Changdong
AU  - Zhang C
FAU - Liu, Mei
AU  - Liu M
FAU - Wang, Xueli
AU  - Wang X
FAU - Chen, Song
AU  - Chen S
FAU - Fu, Xiaojuan
AU  - Fu X
FAU - Li, Geng
AU  - Li G
FAU - Dong, Nianguo
AU  - Dong N
FAU - Shang, Xiaoke
AU  - Shang X
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Altern Ther Health Med
JT  - Alternative therapies in health and medicine
JID - 9502013
RN  - 0 (Toll-Like Receptor 4)
RN  - 60MEW57T9G (alizarin)
RN  - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt)
RN  - 104982-03-8 (Osteocalcin)
RN  - 0 (Core Binding Factor Alpha 1 Subunit)
RN  - 139874-52-5 (NF-KappaB Inhibitor alpha)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 0 (NLR Family, Pyrin Domain-Containing 3 Protein)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Humans
MH  - Osteogenesis/physiology
MH  - Toll-Like Receptor 4/metabolism
MH  - Proto-Oncogene Proteins c-akt/metabolism
MH  - Osteocalcin/metabolism
MH  - Core Binding Factor Alpha 1 Subunit/metabolism
MH  - NF-KappaB Inhibitor alpha/metabolism
MH  - *Aortic Valve Stenosis/metabolism/pathology
MH  - MAP Kinase Signaling System
MH  - Tumor Necrosis Factor-alpha/metabolism
MH  - NLR Family, Pyrin Domain-Containing 3 Protein/metabolism
MH  - Aortic Valve/metabolism/pathology
MH  - *Calcinosis/metabolism/pathology
MH  - Cells, Cultured
MH  - Cell Differentiation/physiology
MH  - Osteoblasts/metabolism
MH  - RNA, Messenger
EDAT- 2022/08/12 06:00
MHDA- 2022/12/27 06:00
CRDT- 2022/08/11 11:14
PHST- 2022/08/12 06:00 [pubmed]
PHST- 2022/12/27 06:00 [medline]
PHST- 2022/08/11 11:14 [entrez]
AID - AT7537 [pii]
PST - ppublish
SO  - Altern Ther Health Med. 2023 Jan;29(1):58-65.