Theoretical And Experimental Study On The Kidney-tonifying Recipes Inducing Proliferation And Differentiation Of Bone Marrow Mesenchymal Stem Cells

Objective:Under the guidance of "kidney essence" theory of Traditional Chinese Medicine, study the intrinsic link between kidney essence and bone marrow mesenchymal stem cells(BMSCs), observe effection of the kidney-tonifying recipes on proliferation and differentiation of BMSCs and Investigate the molecular mechanisms of the kidney-tonifying recipes inducing proliferation and differentiation of BMSCs in order to provide theoretical and experimental basis to explore the scientific meaning of "kidney essence" and the clinical application of the kidney-tonifying recipes.Methods:(1) Theoretical study:through the system study the "Congenital essence" and "kidney essence" theory, and modern research results on stem cell, analysis the "congenital essence" attribute of the stem cells, and the correlation between the mesenchymal stem cells and "kidney essence".(2) Experimental study:eparate and purify BMSCs from SD rats using of bone marrow adherent; by cell morphology, cell growth characteristics, cell cycle, cell surface labeling method of combining test, carriy out on cultured rBMSCs Identification. by MTT detecte effection of Liu Wei Di Huang pills, Shen Qi pills, Zuo Gui pills and You Qui pills on rBMSCs vitro cytotoxicity and proliferation; by Detection of ALP activity,observe effection of water extracts of Liu Wei Di Huang pills, Shen Qi pills, Zuo Gui pills and You Qui pills on rBMSCs osteogenic differentiation; after the intervention of water extracts of the kidney-tonifying recipes, detecte rBMSCs BMP-2 protein expression by Western bloting and Runx2 mRNA expression by RTQ-PCR.Conclusions:(1) Theoretical study:"congenital essence" and stem cells are highly similar on the sources and the functions. as particular correlation between "kidney essence" and mesenchymal stem cells, mesenchymal stem cells can be seen as the material basis of the "kidney essence" functions, the mechanism of the kidney-tonifying recipes treating "kidney essence" deficiency related diseases may be related with kidney-tonifying recipes interventing BMSCs proliferation, differentiation and other biological behavior.(2) experimental studies: ①In the exexperiment, the rBMSCs isolated from the whole bone marrow cells is consistent with the biological characteristics of rBMSCs in vitro established culture system and can meet experimental requirements.②The water extract of Liu Wei Di Huang pills, Shen Qi pills, Zuo Gui pills and You Qui pills can promote the proliferation of cultured rBMSCs in vitro, and tonifying kidney yin recipes is better than inducing the proliferation of rBMSCs than tonifying kidney yang recipes.③The water extract of Liu Wei Di Huang pills, Shen Qi pills, Zuo Gui pills and You Qui pills can promote osteogenic differentiation of cultured rBMSCs in vitro, and in inducing the differentiation of rBMSCs tonifying kidney yang recipes is better than tonifying kidney yin recipes.④water extract of Liu Wei Di Huang pills, Shen Qi pills, Zuo Gui pills and You Qui pills can be increased BMP-2, Runx2mRNA expression of rBMSCs in vitro, suggesting that this may be one of rBMSCs osteogenic differentiation mechanisms.⑤Tonifying kidney yin recipes and tonifying kidney yang recipes show some tendency in inducting proliferation and differentiation of rBMSCs, that is tonifying kidney yin recipes is better than inducing the proliferation of rBMSCs than tonifying kidney yang recipes, and in inducing the differentiation of rBMSCs tonifying kidney yang recipes is better than tonifying kidney yin recipes. It may be the mechanism of action of tonifying kidney yang and tonifying kidney yin produce different therapeutic effects.