| Purpose:According to the the kidney essence marrow main bone "theory,kidney active ingredients of traditional Chinese medicine-inducedosteogenic differentiation of bone marrow mesenchymal stem cellsï¼›clarify the mechanism of action of the kidney Chinese medicine to preventosteoporosis, further kidney essence andmedullary bone viscera theoryprovide experimental basis.Materials and Methods: SPF SD female rats were randomly divided intonormal control group, positive control group (conversion solutiongroup), kidney compatibility group Group1(icariin low-dose, thepsoralen low-dose, oleanolic acid low-dose, stilbene glycosides lowdose), kidney compatibility group2(dose of icariin low, in the doseof psoralen dose of oleanolic acid, stilbene glycosides in dose), kidneycompatibility group3(icariin a low dose, the psoralen high dose, highdose of oleanolic acid, of stilbene glycoside high dose), kidneycompatibility group (in doses of icariin,psoralen low dose,Qioleanolicacid in doses of stilbene glucoside high dose), kidney compatibilitygroup5((in doses of icariin, psoralen medium dose, high dose ofoleanolic acid, stilbene glycoside low dose),kidney compatibility (indoses of icariin, the psoralen high dose, low dose of oleanolic acid,stilbene glycosides in dose), kidney compatibility group7(icariin highdose, Bugulignans low-dose, high dose of oleanolic acid, the stilbene glycosides dose), kidney compatibility group (the icariin high dose ofpsoralen middle dose,low dose of oleanolic acid,stilbeneglycoside highdose), kidney compatibility group of (icariin high dose, high dose ofpsoralen, the dose of oleanolic acid, stilbene glycoside low dose),kidney medicine control group (astragaloside glycosides, ligustrazinegroup)ï¼› by serum pharmacological methods, the experimental ratscontinuous oral administration for3days, drawn (whichever containingserum), the use of rats in each group drug-containing serum-free cultureof mesenchymal stem cells in the bone marrow6days,12days,18daysï¼›calcified nodules were identified by alizarin red stainingï¼› MTT assaybone marrow mesenchymal stem cell proliferation rate, measured by ELISAin osteoblast alkaline phosphatase (Alkaline phosphates, ALP),bonecalcium (osteocalcin, BGP), bone morphogenetic protein2(bonemorphogenetic protein,BMP-2) active expression of gene expression usingquantitative real-time PCR detection of N-terminal kinase1(N-terminalkinase,JNK1).Results:1. Bushen Chinese active ingredients containing serum role of bone marrowmesenchymal stem cells after72hours can promote the proliferation ofbone marrow mesenchymal stem cells cultured in vitro.2. Osteoblasts staining: calcified nodules into a large red, mostlydistributed in the cell density. Negative control, no red, no changein color before.3. Osteoblast ALP enzyme-linked immunosorbent Results: Compared withnormal control group, the kidney of Chinese herbal medicines of9group,positive control group (conversion solution group) and Astragalosidegroup were significantly increased, there was a significant difference(P <0.01)ï¼› compared with the positive control group, Bushen Chinesecompatibility group5,6,8,9group were significantly increased (P <0.01)ï¼›kidney compatibility of Chinese medicine group and the non-kidneyChinese medicine group, Bushen Chinese compatibility group3,4,5,6,7,8,9OD values were significantly higher than the non-kidneyChinese medicine group, there was a significant difference (P <0.01)ï¼›kidney medicine compatibility group comparison between, group5, group6,9OD values were significantly higher than the other groups, therewas a significant difference (P <0.01).4. Osteoblast cells BGP enzyme-linked immunosorbent Results: Comparedwith the normal control group, kidney medicine compatibility group of9group and positive control group (conversion solution group) OD valueswere significantly increased, there was a significant difference (P<0.01)ï¼›positivecontrol group,Bushen Chinese compatibility group group6,9OD values were significantly increased (P <0.01)ï¼› kidneycompatibility of Chinese medicine group and the non-kidney Chinesemedicine group, kidney medicine compatibility group of9groups(orthogonal design) ODvalues were significantly increased, there was asignificant difference (P <0.01)ï¼› comparison between kidney medicinecompatibility group, group5,6groups,9OD value was significantlyhigher than other groups, there was a significant difference (P <0.01).5. Osteoblast cells BMP-2enzyme-linked immunosorbent Results: Comparedwith normal control group, Bushen Chinese compatibility group of9groupand positive control group (conversion solution group), non-kidneyChinese medicine control group OD values were significantly increased,there was a significant difference (P<0.01)ï¼› Bushen Chinesecompatibility group group5,6OD values were significantly increased(P <0.01) compared with the positive control group, Bushen Chinese herbalmedicinesï¼› the kidney medicine group compared with non-kidney Chinesemedicine group, group3,4,5,6,7,8,9OD values were significantlyincreased, there was a significant difference (P <0.01)ï¼› comparison between kidney medicine compatibility group, group5,6,9group) ODvalues were significantly increased, there was a significant difference(P <0.01).6. JNK1: Real-Time PCR results showed: kidney medicine-induced bonemarrow mesenchymal stem cells into osteogenic differentiationexperiments, compared with the normal control group, kidney Chinesemedicine compatibility group a total of9groups and positive controlgroup (conversion of liquid group), non-kidney drugexpression of thecontrol group were significantly increased, there was a significantdifference (P <0.01)ï¼› compared with the positive control group, BushenChinese compatibility group of9groups, non-kidney Chinese medicine thegroup (the Astragaloside group, ligustrazine group) expression levelswere significantly increased(P <0.01)ï¼› compared with the non-kidneyChinese medicine group (TMP group), kidney medicine compatibility group(group4) expression levels were significantly increased, there was asignificant difference (P <0.01)ï¼› with Astragaloside group,kidneyChinese herbal medicines group (Group4,group5) expression levelswere significantly increased, there was a significant difference (P<0.01).Conclusions:1. kidney active ingredients of traditional Chinese medicine can promotebone marrow mesenchymal stem cells to differentiate into osteoblasts,improve osteoblasts value-added rate.2. Mechanism of bone marrow mesenchymal stem cells to differentiate intoosteoblasts and JNK1signaling pathway. |
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