title: [Influence of exogenous nitric oxide on chondrogenic differentiation of adipose derived stem cells].
abstract: OBJECTIVE
  To determine the effects of exogenous nitric oxide (NO) donor sodium nitroprusside (SNP) on the proliferation and expression of related-gene of adipose-derived stem cells(ADSCs) and its role in chondrogenic differentiation of ADSCs.
  
  
  METHODS
  Rat ADSCs were harvested and cultured, and then induced to osteogenic and adipogenic differentiations, detected with Alizarin red stained and Oil red O stain, respectively. The change of NO during chondrogenic differentiation of ADSCs was tested by NO detection kit. Cell counting kit-8 (CCK8) was used to detect the proliferation of ADSCs under different concentrations of SNP (0.25 mmol/L, 1.00 mmol/L, 4.00 mmol/L). Gene expression level of transformation growth factor (TGF-beta1) and specific gene of chondrogenic differentiation-signaling protein Smad3 and Collage II alPHA (Col-II alpha1), were detected by Real time- PCR (RT-PCR) method.
  
  
  RESULTS
  Positive alizarin red staining and Oil red O staining were found after osteogenic and adipogenic induction of cultured ADSCs. Higher concentrations of NO were found in the supernatant of the experimental group with ADSCs chondrogenic differentiations compared with the controls (P<0.05). Low concentrations (0. 25 mmol/L, 1.00 mmol/L) of SNP showed no significant effects on cell proliferations (P>0.05), whereas high concentration (4. 00 mmol/L) of SNP inhibited cell proliferation (P<0.05). RT-PCR revealed that SNP inhibited the gene expression of TGF-beta1 mRNA and chondrogenic differentiation of specific gene Smad3 mRNA, Col-II alpha1 mRNA.
  
  
  CONCLUSIONS
  SNP can inhibit chondrogenic differentiations by suppressing the production of TGF-beta1 and inhibiting downstream of TGF-beta1 signaling pathways, thereby inhibiting ADSCs differentiation into chondrocytes.
authors: Shuang
authors: S
authors: Liu
authors: Hai-sen
authors: HS
authors: Huang
authors: Mei
authors: M
authors: Yu
authors: Xiao-rin
authors: XR
authors: Luo
authors: Wei-hua
authors: WH
authors: Guo
authors: Wei-dong
authors: WD
authors: Tian
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects; D024043:Collagen Type II / Q000378:metabolism; D009569:Nitric Oxide / Q000494:pharmacology*; D009599:Nitroprusside / Q000494:pharmacology; D051381:Rats; D051900:Smad3 Protein / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D024043:Collagen Type II; D051900:Smad3 Protein; C497787:Smad3 protein, rat; C508962:Tgfb1 protein, rat; D053773:Transforming Growth Factor beta1; D009599:Nitroprusside; D009569:Nitric Oxide