Effect Of Protoscolex Of Echinococcus Alveolaris On Calcification Of Bone Marrow Mesenchymal Stem Cells

Objective Under the condition of co culture of Echinococcus granulosus protoscolex and Echinococcus vesiculatum protoscolex with mouse bone marrow mesenchymal stem cells in common medium and osteogenic differentiation induction medium,the expression of the two kinds of Echinococcus protoscolex on the related calcification factors of mouse bone marrow mesenchymal stem cells was compared,and the mechanism of mesenchymal stem cells involved in the formation of calcification focus of Echinococcus was preliminarily discussed.Methods Extract and culture the protoscolex of Echinococcus granulosus and Echinococcus vesiculatum;Extraction,culture and identification of MSCs;The bone marrow mesenchymal stem cells and the protoscolex of two Echinococcus species were cultured in common medium;Bone marrow mesenchymal stem cells and protoscolex of two Echinococcus species were co cultured under the condition of osteogenic differentiation medium;The alkaline phosphatase activity of MSCs was detected by micro enzyme labeling;Western blotting was used to detect the expression of BMP2,RUNX2,p-smad1 / 5 / 8 and ?-actin in MSCs of each group;RT-q PCR was used to evaluate the m RNA expression of BMP2,OCN and RUNX2.Results ALP activity test results: under normal culture conditions,ALP activity of the MSc + CE group and the MSc + AE group were higher than that of the control group(P < 0.05),and ALP activity of the MSc + CE group was higher than that of the MSc + AE group on the 1st,4th and 7th day(P < 0.05).The condition of osteogenic induction culture: the ALP activity expression of Echinococcus granulosus protoscolex co culture group was higher than that of Echinococcus vesiculosus protoscolex co culture group(P < 0.05).Western blotting showed that the expression of RUNX2 protein in Echinococcus granulosus model group and Echinococcus vesiculosa model group was higher than that in the blank control group(P <0.05);the expression of BMP 2,RUNX2 and p-smad1 / 5 / 8 protein in the MSc + CE group and the MSc + AE group on the 1st and 4th day was higher than that in the control group(P <0.05),and that in the MSc + CE group on the 1st,4th and 7th day was higher than that in the control group(P < 0.05)BMP2,RUNX2,p-smad1 / 5 / 8 were higher than those of MSC +AE group(P < 0.05);osteoblasts induced differentiation culture conditions: the expressions of BMP2,RUNX2,p-smad1 / 5 / 8 in MSCs group were significantly higher than those in Echinococcus vesiculatum and Echinococcus granulosus(P < 0.05).The results of RT-q PCR showed that the RUNX2 m RNA level of Echinococcus granulosus and Echinococcus alveolaris models was higher than that of the blank control group(P < 0.05);the general culture conditions: the expression of RUNX2 m RNA from high to low on the 1st,4th and 7th day in MSC + CE group,MSC + AE group and control group(P < 0.05);the expression of BMP 2 m RNA in the 1st,4th and 7th day in MSC + CE group was higher than that in MSC +AE and control group(P < 0.05),while that in the 1st and 4th day in MSC + AE group was higher than that in MSC + AE and control group(P < 0.05)There was no significant difference in the expression of BMP 2 m RNA(P > 0.05);the condition of osteogenic differentiation culture: the expression level of BMP 2,RUNX2,OCN and Smad1 m RNA in MSCs cultured for 7 days was low(P < 0.05);the expression level of RUNX2 m RNA in the protoscolex co culture group of Echinococcus granulosus was significantly higher than that in the protoscolex co culture group(P < 0.05).conclusion1.The occurrence and development of liver mineralization around the focus of echinococcosis are related to its natural course of disease;2.The protoscolex of Echinococcus vesiculatum and Echinococcus granulosus both promote the early calcification of mesenchymal stem cells,and the protoscolex of Echinococcus granulosus is more effective;3.The osteogenic differentiation of mesenchymal stem cells can be inhibited by protoscolex of Echinococcus.The mechanism may be related to the biphasic regulation of protoscolex of Echinococcus on the calcification of mesenchymal stem cells.