| Objective To observing the mice after two foot swelling,morphological changes and cells induce osteogenesis in the film,study of BuShenShuJi granule(BSG)through the BMP pathway delay mechanism of heterotopic ossification in ankylosing spondylitis with DBA/1 male mouse and human bone marrow mesenchymal stem cells for research.Methods 1.Animal experiments: the weight 25 ± 5g,DBA/1 male mice of 12 weeks were randomly divided into two doses in the high-dose group,middle dose group and low dose groups,the Western Group and group,each group of 8.When a week for 14 weeks in mice,were given drugs in mice,mice in the 20,22,24 weeks to observe the two-foot swelling,swelling score and statistics.In the weeks after the 24 th week of taking double mouse feet and heel ligament attachment parts of pathological specimens,HE stained light microscopic observation on pathological morphology changes.2.Cell experiment: the human BMSCs primary cells passaged amplification to the 4th generation,planted directly in six-well plates,T-25,flask,randomly divided into high dose,middle dose and low dose groups,the Western Group and control group.Then use the above cell culture mesenchymal stem cells culture medium to90% above,you again use induction induced by serum containing medium and culture,28 days after the extraction of cellular RNA and protein expression and detection of the target factor.Result 1.animal experiment: the comparison of mouse foot swelling of mice in each group: after 14 weeks of age,were gradually toe swelling,redness,stiffness and deformation performance of toe toe.With the increase of the age of mice,the degree of swelling increased gradually.In mice at twentieth weeks were statistically significant difference between the groups(P<0.05),in which the Chinese medicinehigh dose group than in low dose group,model group,the degree of swelling of light(P<0.05);Chinese medicine dose group than the model group the degree of swelling of light(P<0.05).In mice at twenty-second weeks were statistically significant difference between the groups(P<0.05),Chinese medicine high dose group and middle dose group were lighter than the model group(P<0.05);the degree of swelling in mice at twenty-fourth weeks were statistically significant difference between the groups(P<0.05),in which the Chinese medicine high dose group,middle dose group and Western medicine group compared with the model group the degree of swelling of light(P<0.05).In the medium dose mice,the swelling degree of the 24 week old is heavier than that of the 20 week old(P<0.05).The double front foot and heel ligament after comparison of adhesion site pathological score: compared with the model group,the kidney Shu ridge granules in each dose group and Western medicine group were significantly lower than those of the model group,the difference was statistically significant(P<0.05).2.cell experiments: 1 the comparison effect of Bushen Granule on BMP-4 factor of Shu ridge: compared with the model group,the level of mRNA in western medicine group increased(P<0.05);the expression level in the high dosage group,mRNA and protein decreased,with statistical significance(P<0.05).Compared with the western medicine group,the expression of mRNA and protein in the middle dose group of Chinese medicine decreased,which was statistically significant(P<0.05).The effect of Bushen Granule on the Smad-4 ridge Shu factor: compared with model group,Chinese medicine high dose group,middle dose group and low dose group mRNA and protein expression level decreased,with statistical significance(P<0.05).Compared with the western medicine group,the levels of mRNA and protein in the high,medium and low dosage groups of Chinese medicine decreased,and the difference was statistically significant(P<0.05).The effect of Bushen Granule on the Runx-2 ridge Shu factor: effect of Bushen Granule on Runx-2 mRNA Shu ridge: compared with the model group,low dose group and mRNA protein expression in the low-dose group of Chinese medicine and traditional Chinese medicine,with statistical significance(P<0.05).Compared with the western medicine group,the expression of mRNA and protein in the low,medium and high dosage groups ofChinese medicine decreased,which was statistically significant(P<0.05).Conclusion 1.BSG high dose,dose and Western medicine can be in varying degrees of improvement of DBA/1 mice after two foot swelling,and high-dose to prolong two-foot swelling after.After Western medicine can prolong double foot swelling,but onset time than BSG materials.2.Pathology: kidney-tonifying granule BSG group and Western medicine can be improved in varying degrees DBA/1 double rear foot tissue morphology in mice,but BSG no significant difference compared with Western medicine.3.Kidney-tonifying granule dosage of BSG to different extent,BMP-4,Smad-4 and Runx-2 the expression,indicating kidney BSG can be Heterotopic ossification BMP pathway delays with ankylosing spondylitis. |
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