Studies On Novel Cell Membrane Chromatography And The Active Compounds Screening Of The Compatibility Of Ploygonatum Sibiricum And Coptis Chinensis

Objective: To improve the technology of CMC, the stability of it was studied.To expand the application of CMC in the evaluation of the active compounds in anti-diabetes of natural product, this work try to set up novel CMCs. And the active compounds of the combination of Coptis chinensis and Polygonatum sibiricum was evaluated.Methods: 1.The test of glucose consumption by insulin-resistant 3T3-L1 adipocytes was used to confirm the therapy effect of the combination. 2.The stability of cell membranes was evaluated by the variation of the index of ATPase.3.The active compounds such as glibenclamide,berberine were used for develop the pancreatic islet cells membrane chromatography, HepG2 cell membrane coated magnetic bead fishing chromatography and 3T3-L1 adipocytes chromatography.4.The test of glucose consumption by insulin-resistant 3T3-L1 adipocytes was employed for confirm the screening result of these chromatography managements.Results: 1. The pharmacological tests showed the glucose consumption of the extract treated insulin resistant 3T3-L1 adipocytes were increased significantly(P<0.01). 2. The cell membrane stability study showed BSA and NaCl solution could improve the stability of ATPase. So they could increase the stability of cell membrane and so is that of CMC. 0.9% NaCl solution and 1% BSA solution could reduce the degradation of ATPase which is lying on the surface of cell membrane in0-9h. 3. The evaluation result of cell membrane adsorption isotherm profile and adsorption time-depend profile indicate that the optimal cell membrane protein coating technology is about 1.2 mg/mL for 6 h. 4. Pancreatic islet cell membrane was coated on the surface of capillary column. The pancreatic islet cell membrane capillary column was set up. This management could reduce the consume of thecellular target cell. For the improvement of the resolution of the chromatography, the pancreatic islet cell membranes were coated on the surface of magnetic beads. And we called it as cell membrane coated magnetic beads fishing. The active compounds such as glibenclamide were fished. By this method, we found berberine and other 5compounds in Coptis chinensis and three homoisoflavanones in Polygonatum sibiricum. And management could also be used for the active compounds screening of natural products. Comparisons of the developed management with the CMC method also showed the noteworthy advantages of the fishing technique regarding the usage of cellular membranes, buffers as well as length of operation time. 5. For screening the active compounds which may interactive with signal passway of cells,insulin resistant 3T3-L1 adiposities and insulin resistant HepG2 cell were used for screening such compounds. These cells were cultured and were used to extract the active compounds of combination. By the models we found berberine and other compounds in Coptis chinensis and 6,8-dimethyl-4’,5,7-trihydroxyl homoisoflavanone and other compounds in Polygonatum sibiricum.Conclusion: 3T3-L1 adipocytes extract, HepG2 cell extract and islet cell membrane coated magnetic beads could be used for extract active compounds in natural products. In the combination group, the active compounds were the alkaloids which contented by Coptis chinensis and the homoisoflavanones which contented by Polygonatum sibiricum. As the reasons of the compounds structure and the targets may different. Activity of this combination may be more efficient than that of the extracts of two the plants which extracted separately. And these managements may not only be used for the screening for other natural products, but also could provide the basis for the compatibility.