| Purpose:By observing the effect of continuous gluten bone tablet on bone formation in fracture rats and the activation of BMP-2/Smad pathway and TGF-? / SMAD pathway in callus,the mechanism of TGF-? superfamily regulation was studied.? Material and method:After one week of adaptive feeding,50 SD rats were randomly divided into two groups: sham-operated group,model control group,high-dose group,low-dose group,Xianling Gubao group,10 rats in each group.The model of fracture was duplicated in the model control group,high dose group,low dose group and Xianling Gubao group.Sham-operated rats were operated on without fracture.Drug intervention was given once a day for 3 weeks on the fifth day after operation.1 hour after the last administration,whole blood was collected,serum was isolated and serum Ca,P,ALP,BGP content was detected.The callus tissue attached to the diaphysis was used to detect the expression of SMAD1/5,SMAD6 in callus tissue by western-blot.The remaining femoral tissue was immersed in 4% paraformaldehyde solution and fixed at room temperature.The expression of TGF-? and BMP-2 in callus was detected by HE staining and immunohistochemistry? Results: 1.The pathological section of femur in each group showed that the bone structure was clear and the bone plate structure was normal in sham-operated group.In the model control group,the bone tissue was damaged,the bone plate was incomplete,a large number of inflammatory cells were infiltrated,osteoblasts were more,and osteoclasts were occasionally seen.The pathological changes of the rats in the high,low dose and Xianling Gubao groups were significantly lighter than those in the model control group,and the inflammatory cells were decreased significantly.A large number of osteoblasts and osteoclasts were observed.2.The results showed that compared with the sham operation group,the serum calcium and phosphorus contents in the other groups were significantly decreased(p < 0.01).The results showed that the contents of calcium and phosphorus in the serum of the rats in each group were significantly lower than those in the sham operation group(p < 0.01).Compared with the model control group,the levels of serum calcium and phosphorus in the high dose group and Xianling Gubao group were significantly higher than those in the model control group,while the serum phosphorus content in the low dose group also increased significantly(p < 0.01).Compared with the high dose group,the serum calcium and phosphorus content in the low dose group was significantly lower than that in the Xianjinjiegu tablet group,while the serum phosphorus content in the Xianling Gubao group was significantly lower than that in the Xianjinjiegu tablet group(p < 0.05 or 0.01).3.The results of ALP,BGP detection in serum of rats in each group showed that compared with sham operation group,the content of ALP,BGP in serum of other groups was significantly lower than that of sham operation group(p < 0.01).Compared with the model control group,the serum ALP,BGP content in the drug intervention group was significantly higher than that in the model control group(p < 0.01).Compared with the high dose group,the content of serum ALP,BGP in the low dose group and the content of serum BGP in Xianling Gubao group were significantly lower than those in the high dose group(p < 0.01 or 0.05).Compared with the low dose group,the serum BGP content of Xianling Gubao group was significantly higher than that of the low dose group(p < 0.05).4.The expression of SMAD1/5,SMAD6 in callus of rats in each group was detected.The results showed that the band of SMAD1/5 protein appeared at 120 kDa and 58 kDa after luminescence,and the band of SMAD6 protein appeared at 54 kDa.Compared with the sham-operated group,the expression of SMAD1/5 protein decreased significantly in the other groups,but the expression level of SMAD6 protein increased significantly(p < 0.01).Compared with the model control group,the expression of SMAD1/5 protein increased significantly in the skeleton of rats in each drug intervention group,but the expression level of SMAD6 protein decreased significantly(p < 0.01).Compared with the high dose group,the expression of SMAD1/5 protein in callus of rats in low dose group and Xianling Gubao group was significantly decreased,while the expression level of SMAD6 protein was significantly increased in the low dose group and Xianling Gubao group.There was statistical difference(p < 0.01).Compared with the low dose group,the expression of SMAD1/5 protein in the callus of Xianling Gubao group was significantly increased,while the expression level of SMAD6 protein was significantly decreased(p < 0.01).Conclusion: 1.It can promote the healing of fracture rats and increase the contents of calcium,phosphorus,alkaline phosphatase and osteocalcin in the serum of fracture rats.Reduce the inflammatory infiltration in the callus of fracture rats,increase osteoblasts and so on.2.The effect of bone graft on bone formation in fracture rats may be achieved by promoting the activation of TGF-? / Smad,BMP/Smad signaling pathway.Its core mechanism may be the regulation of TGF-? superfamily by continuous gluten bone tablet. |
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