title: Use of small interfering ribonucleic acids to inhibit the adipogenic effect of alcohol on human bone marrow-derived mesenchymal cells.
abstract: This study tested the potential of small interfering RNAs (siRNA) targeting human peroxisome proliferator activated receptor gamma (PPARγ) to repress the adipogenic effect of alcohol on human bone marrow-derived mesenchymal cells (hBMSCs). hBMSCs were cultured from hip replacement surgery patients (n = 10). PPARγ-siRNA was transiently transfected into hBMSCs cultured in ostogenic media containing 50 mM alcohol by using a liposome-based strategy. Oil red O staining was used to test the development of differentiated adipocytes, and Alizarin red staining was used to test mineral deposition. Marker genes of adipogenesis (PPARγ2 and aP2) and osteogenesis (Osf2/Cbfa1) were examined through real time RT-PCR and Western blot, respectively. Collagen type I, alkaline phosphatase and osteocalcin protein synthesis of cultures were also assayed. Data were presented as mean ± SD. Differences between the means of the treatment groups were determined with ANOVA. PPARγ-siRNA transfection resulted in significantly lower adipocyte number, increased matrix mineralisation, repressed adipogenic gene markers, up-regulated osteogenic gene marker and bone matrix protein synthesis in the PPARγ-siRNA group compared to controls (P < 0.05). PPARγ-siRNA is a useful strategy to inhibit the adipogenic effect and the osteogenic repression of alcohol on hBMSCs. This may be a novel therapeutic intervention for osteopenic disorders in alcoholism and other conditions.
authors: Department of Orthopedics, West China Hospital, Sichuan University, Sichuan, China.
authors: Qiang
authors: Q
authors: Huang
authors: Hui
authors: H
authors: Zhang
authors: Fu-xing
authors: FX
authors: Pei
authors: Zhi-yu
authors: ZY
authors: Chen
authors: Guang-lin
authors: GL
authors: Wang
authors: Bin
authors: B
authors: Shen
authors: Jing
authors: J
authors: Yang
authors: Zong-ke
authors: ZK
authors: Zhou
authors: Qing-quan
authors: QQ
authors: Kong
mesh_terms: D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D050156:Adipogenesis / Q000187:drug effects* / Q000235:genetics; D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D000431:Ethanol / Q000633:toxicity*; D050556:Fatty Acid-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D005819:Genetic Markers; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008875:Middle Aged; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D047495:PPAR gamma / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
chemical_list: D015415:Biomarkers; D015815:Cell Adhesion Molecules; D050659:Core Binding Factor Alpha 1 Subunit; C493410:FABP4 protein, human; D050556:Fatty Acid-Binding Proteins; D005819:Genetic Markers; C512835:POSTN protein, human; D047495:PPAR gamma; D034741:RNA, Small Interfering; D000431:Ethanol