Use of small interfering ribonucleic acids to inhibit the adipogenic effect of alcohol on human bone marrow-derived mesenchymal cells |
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Authors: | Qiang Huang Hui Zhang Fu-xing Pei Zhi-yu Chen Guang-lin Wang Bin Shen Jing Yang Zong-ke Zhou Qing-quan Kong |
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Institution: | (1) Department of Orthopedics, West China Hospital, Sichuan University, Sichuan, China;(2) Department of Anesthesiology, Xinhua Hospital affiliated Shanghai University Medical College, Shanghai, China; |
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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. |
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