Use of small interfering ribonucleic acids to inhibit the adipogenic effect of alcohol on human bone marrow-derived mesenchymal cells

• Published in: International orthopaedics 2010-10, Vol.34 (7), p.1059-1068
• Main Authors: Huang, Qiang, Zhang, Hui, Pei, Fu-xing, Chen, Zhi-yu, Wang, Guang-lin, Shen, Bin, Yang, Jing, Zhou, Zong-ke, Kong, Qing-quan
• Format: Article
• Language: English
• Subjects: Acids; Adipocytes; Adipocytes - drug effects; Adipocytes - metabolism; Adipocytes - pathology; adipogenesis; Adipogenesis - drug effects; Adipogenesis - genetics; Adult; Alcoholism; Alkaline phosphatase; Biomarkers - metabolism; Bone marrow; Bone Marrow Cells - cytology; Bone matrix; Bone surgery; Cbfa-1 protein; Cell Adhesion Molecules - genetics; Cell Adhesion Molecules - metabolism; Cell Count; Cell culture; Cell Differentiation - drug effects; Collagen (type I); Core Binding Factor Alpha 1 Subunit - genetics; Core Binding Factor Alpha 1 Subunit - metabolism; Data processing; Drug abuse; Ethanol; Ethanol - toxicity; Fatty Acid-Binding Proteins - genetics; Fatty Acid-Binding Proteins - metabolism; Gene Expression Regulation - drug effects; Genetic Markers; Hip; Humans; Medicine; Medicine & Public Health; Mesenchymal Stromal Cells - drug effects; Mesenchymal Stromal Cells - metabolism; Mesenchymal Stromal Cells - pathology; Mesenchyme; Middle Aged; Mineralization; Minerals; Original Paper; Orthopedics; Osteocalcin; Osteogenesis; Osteogenesis - drug effects; Osteogenesis - genetics; Peroxisome proliferator-activated receptors; Polymerase chain reaction; PPAR gamma - genetics; PPAR gamma - metabolism; Protein biosynthesis; RNA, Small Interfering - genetics; siRNA; Therapeutic applications; Transcription factors; Transfection; Western blotting
• ISSN: 0341-2695; 1432-5195
• DOI: 10.1007/s00264-009-0914-y

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