MicroRNA-138 Suppresses Adipogenic Differentiation in Human Adipose Tissue-Derived Mesenchymal Stem Cells by Targeting Lipoprotein Lipase

PURPOSE: Adipogenic differentiation of adipose tissue-derived mesenchymal stem cells (AMSCs) is critical to many disease-related disorders, such as obesity and diabetes. Studies have demonstrated that miRNA-138 (miR-138) is closely involved in adipogenesis. However, the mechanisms affected by miR-138 remain unclear. This work aimed to investigate interactions between miR-138 and lipoprotein lipase (LPL), a key lipogenic enzyme, in AMSCs. MATERIALS AND METHODS: Human AMSCs (hAMSCs) isolated from human abdomen tissue were subjected to adipogenic differentiation medium. Quantitative real-time polymerase chain reaction and Western blot assay were applied to measure the expressions of miR-138, LPL, and the two adipogenic transcription factors cytidine-cytidine-adenosine-adenosine-thymidine enhancer binding protein alpha (C/EBPα) and peroxisome proliferator-activated receptor gamma (PPARγ). The relationship between miR-138 and LPL was predicted utilizing the miRTarBase database and validated by dual luciferase reporter assay. RESULTS: Showing increases in C/EBPα and PPARγ expression levels, hAMSCs were induced into adipogenic differentiation. During adipogenesis of hAMSCs, miR-138 expression was significantly downregulated. Overexpression of miR-138 by transfection inhibited hAMSCs adipogenic differentiation in vitro. Mechanically, LPL was a target of miR-138. LPL expression was upregulated during adipogenesis of hAMSCs, and this upregulation was reversed by miR-138 overexpression. Functionally, silencing of LPL by transfection exerted similar inhibition of the expressions of C/EBPα and PPARγ. Meanwhile, LPL ectopic expression was able to partly abolish the suppressive effect of miR-138 overexpression on adipogenic differentiation of hAMSCs. CONCLUSION: Upregulation of miR-138 inhibits adipogenic differentiation of hAMSCs by directly downregulating LPL.

In vivo tracing of human amniotic mesenchymal stem cells labeled with PKH26 in rat intrauterine adhesions model / 生物工程学报

To observe the migration of human amniotic mesenchymal stem cells (hAMSCs) labeled with PKH26 in the endometrium of rats intrauterine adhesion. hAMSCs were isolated, identified and labeled with PKH26 to detect the biological characteristics of the cells. Rat intrauterine adhesion models were established using mechanical and infective method and PKH26-labeled hAMSCs were transplanted through the tail vein. The distribution of PKH26 labeled hAMSCs in the endometrium of rats were observed with the fluorescence confocal microscope. The results showed that PKH26 stain had no significant effect on cell activity, cycle, apoptosis and so on. PKH26-labeled positive cells were mainly distributed in injured endometrium of rats. It shows that the PKH26 labeling technique is a safe and effective method for tracing the human amniotic mesenchymal stem cells in the treatment of intrauterine adhesions.