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Objective To study the mechanism of differentiation of high glucose induced M2 macrophages used by high glucose medium stimulation. Methods Raw264.7 (murine macrophage cell line)was cultured and stimulated by 4. 25% glucose medium, and mannitol medium was used as osmotic pressure control. Cells were harvested at 0h,4h,8h and 12h to examine the expression of CD206 and Arg-1 and the activation of PI3K/Akt signaling pathway. After blocking the PI3K/Akt signaling pathway by LY294002(the specific inhibitor of PI3K),the expression of Arg-1 was examined by western blot. Results The expression of Arg-1 was increased from 8h while CD206 reached the peak at 12h induced by high glucose which indicated the activation of M2 in a time-dependent manner. Akt was phosphorylated at 8h stimulated by high glucose medium. The specific inhibitor of PI3K reduced the expression of Arg-1 by blocking the phosphorylation of Akt. Conclusion High glucose, rather than high osmotic pressure,induced macrophages polarized into M2 via activation of PI3K/Akt signaling pathway.
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Objective To establish a HPLC method for determination of madecassoside and asiaticoside in Centella asiatica formula granules.Methods Chromatographic separation was performed on Ultimate AQ-C18 column(4.6 mm×250 mm,5 μm) with mobile phase of acetonitrile(A)-2 mmol/L β-cyclodextrin(0~30 min:21% A→23% A;30~60 min:23% A→25% A).The flow rate was set at 1.0 ml/min, the column temperature at 30 ℃ and detection wavelength at 205 nm.Results Madecassoside and asiaticoside showed good linearity (r>0.9995) in the ranges of 0.187 7~3.754 μg and 0.184 3~3.686 μg respectively.The specificity, repeatability, precision,recovery and stability were satisfied to the method validation requirements of China Pharmacopoeia.Conclusion The method can determine madecassoside and asiaticoside in Centella asiatica formula granules.
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BACKGROUND:The complex relationship between bone marrow mesenchymal stem cels and cancers severely limit the clinical application of mesenchymal stem cels. So it is urgent to study the role of mesenchymal stem cels in tumor growth and metastasis. OBJECTIVE:To explore the effect of human bone marrow mesenchymal stem cels on epithelial mesenchymal transition in non-smal cel lung cancer A549 and PAa cels. METHODS:The A549 and PAa cels were cultured with mesenchymal stem cel supernatant (mesenchymal stem cel conditioned medium, MSCs-CM). The celular morphology was observed under a microscope. The mRNA and protein expression of E-cadherin, N-cadherin, Vimentin, Slug, Snail, and Twist were determined by RT-PCR and western blot. Transwel and wound healing assay were used to detect the change of migration and metastatic ability. RESULTS AND CONCLUSION:Compared with the control group, the celular morphology of experimental group showed mesenchymal-like changes. In response to MSCs-CM, there was decreased E-cadherin but increased N-cadherin, Vimentin and Slug, Snail, Twist at mRNA and protein levels compared with the control group (P < 0.05). The migration and metastatic abilities of the experimental group were also increased. So, human bone marrow mesenchymal stem cels can promote epithelial mesenchymal transition in A549 and PAa cels, and enhance the migration and metastatic abilities of A549 and PAa cels.