Effect Of Bushen Huoxue Capsule Medicated Serum On Glucocorticoids Intervention Of Rat Bone Marrow Stromal Cell Proliferation And Differentiation

Objective: To investigate the effect of Bushen Huoxue Capsule medicated serum on glucocorticoids intervention of rat bone marrow stromal cell proliferation and differentiation.Method: 1.Preparation of different concentrations of serum containing Bushen Huoxue capsules were healthy male Wistar rats were divided into four groups: low dose group,middle dose group,high dose group,negative control group.According to the formula Meeh-Rubner A(surface area)=K(coefficient).W(weight g)2/3.10-4,calculate in traditional Chinese medicine Bushen Huoxue Capsule dose of 14.5g/kg.d,2g/ml and 4g/ml to self~decocting,bottle containing 4 degrees cold standby.The control group were given normal saline,low dose group,middle dose group,high dose group three groups according to the ratio 1:2:4(14.5g/kg.d,29g/kg.d,58g/,kg.d)were administered continuously intragastric administration for 10 days.The rats were in the sky after eleventh abdominal 6h sodium pentobarbital intraperitoneal injection anesthesia of inferior vena artery blood,into the test tube after solidification,immediately the specimen at a temperature below 4 DEG 3500r/min centrifugal,centrifugal 10 min,learn from the upper serum,-80 C storage backup;2.Isolation,culture and passage of Wistar male rat bone marrow stromal cells(BMSCs),the growth and morphology of BMSCs,to the 3 generation cells for experiment.BMSCs adding 1×10-6 mmol/L dexamethasone sodium phosphate hormone single factor simulation damage model;3.Different concentrations of Bushen Huoxue Capsule medicated serum on BMSCs and prognosis of dexamethasone sodium phosphate,affecting the value and activity detection of BMSCs MTT method to detect BMSCs cell cycle;flow cytometry;alkaline phosphatase staining with calcium cobalt method,alizarin red staining and calcium nodules,immunohistochemical staining of type I collagen in detection of BMSCs the effect of osteogenic differentiation.Result: 1.There was no significant difference in the activity of BMSCs and the activity of BMSCs after hormone intervention with different concentration of Bushen Huoxue capsule.The effect of dexamethasone sodium phosphate on differentiation of BMSCs into osteoblast was obvious,especially in the middle and high dose groups,the effect of hormone on the osteogenic differentiation of bone marrow stromal stem cells was obvious,the low dose group and the low dose group had no obvious cytotoxicity;And the control group had no significant difference.3.The effect of low-dose serum containing drugs on BMSCs DNA replication was the most obvious.With the increase of concentration,Bushen Huoxue Capsule reduced the DNA replication ability of BMSCs.After intervention of BMSCs with different concentrations of Bushen Huoxue capsules,the DNA replication ability of BMSCs was improved,but there was no significant difference.Conclusion: 1.Bushen Huoxue Capsule can inhibit the apoptosis of BMSCs,and promote its value-added activity,and no cytotoxicity;2.Bushen Huoxue capsule containing serum can promote the differentiation of BMSCs into osteogenic direction after single injury of Glucocorticoids.