Study On The Expression Of GPR48 And Related Factors In Rat BMSCs By Serum Containing Yougui Pill

Purpose:To explore the mechanisms of prophylaxis and treatment of osteoporosis due to deficiency of kidney with Yougui pill,based on the TCM theory of “the kidney stores the essence,produces the marrow and controls the bones”.To observe the effect of Yuogui Pill containing serum to the differentiation from bone marrow mesenchymal stem cells to osteoblast and the expression of GPR48、ATF4、BSP、BMP7、TGF-βand BGP in cells of rats through vitro experiment and provides scientific bases for clinical treatment of osteoporosis.Material and method: The drug-contained serum was prepared using serum-pharmacological method in this experiment.Forty 3-month-old unmated SD female rats of SPF grade,which weighted 190-220 g were randomly divided into normal group,induced group,Yougui Pill group and Fosamax group,Gushukang group,8 rats in each group.The rats were treated with experimental drugs for one week continuously,once-daily,and the rats were killed after one week.Took the corresponding tissue and keep the drug-containing serum of each group.The bone marrow mesenchymal stem cells were isolated and extracted from the femur and tibia,and were cultured and purified.MMT assay was used to observe cell proliferation,the protein expression of ALP、GPR48、ATF4、BSP、BMP7、TGFβ、BGP in cells were detected by ELISA.Results:1.MTT results of each experimental group showed: comparing with the normal group,the expression of Yougui Pill group significantly increased(P<0.01);comparing with the induced group、Gushukang group、fushanmei group,Yougui Pill group was significantly different(P<0.01).2.Serum ALP activity results of each group showed: comparing with the normal group,the expression of Yougui Pill group was significantly different(P<0.01);comparing with the induced group、Gushukang group、fushanmei group,Yougui Pill group was significantly different(P<0.01).3.The protein expression of GPR48,ATF4,BSP,BMP7,TGF-β and BGP in rat bonemarrow mesenchymal stem cells of each group showed that comparing with the normal group,the other four groups were dramatically improved(P<0.01).Conclusion:1.GPR48,ATF4,BSP,BMP7,TGF-βand BGP can be expressed in normal rat bone marrow mesenchymal stem cells,indicating that GPR48,ATF4,BSP,BMP7 and TGF-beta signal factors were present in rats.2.Yuogui Pill containing serum can improve the protein express of GPR48,ATF4,BSP,BMP7,TGF-βand BGP in rat bone marrow mesenchymal stem cells,which means Yougui Pill may be achieve the purpose of preventing and treating osteoporosis by adjusting the protein expression level of GPR48,ATF4,BSP,BMP7,TGF-βand BGP.3.Yougui Pill containing serum can promote rat bone marrow mesenchymal stem cells to differentiate into osteoblasts,the interpretation of the traditional Chinese medicine "kidney essence,scientific bone marrow" theory.It explains the scientificalness of the TCM theory of“the kidney stores the essence,produces the marrow and controls the bones”.