The Impact Of The Serum Of Kidney-tonifying Chinese Herbs On The Bmp-smad Signal Transduction Pathways In Osteoblasts Studies

Purpose:Through studying the impact of Kidney-tonifying Chinese herbs in osteoblasts to the act- ive change of BMP2,BMP7 and Smad1/5 in SD rats, explore the osteoporosis pathogenesis mechanism of molecular biology and study the mechanism of action of herbs prevention oste- oporosis to provide scientific experiment basis for the clinical prevention of osteoporosis.Material and method: To get freshmen SD rats' cranial bones of osteoblast through co-llagenase digestion and cultivate it in vitro and identify the alkaline phosphatase expression with cobalt calcium of appraisal. The experimental rats were randomly divided into normal group, Gushukang parti- cle control group (Gushukang group), Kidney-tonifying Chinese herbs group (Kidney-tonif- ying group) and Tonifying the Middle and Replenishing group (tonifying spleen group). Eight days after the medicine intervention of experimental rat, the serum of each group's rats was used to cultivate the osteoblasts in rats for 48 hours.Besid- es, there were comparison groups with Protease inhibitors MG132 of UPP pathway. We tested the appreciation rate of vitro ost- eoblasts with MTT and the activity expressions of BMP2, BMP7, Smad1/5 with Immunohist- ochemistry SABC.Results:1 48 hours after the kidney medicine serum applied to the vitro osteoblasts, the vitro osteobla- sts can obviously promote the proliferation of osteoblasts (P < 0.01).2 The application of UPP pathway inhibitor MG - 132 protease body can obviously inhibit the proliferation of osteoblast (P < 0.01).3 The results of the dyeing of alkaline phosphatase: osteoblast via the improved Gomori Calc- uim Cobail method and the redyeing of hematoxylin, some brownish-black fine particles can be seen mostly on and around the epicytes, and a few in the endochylema. There are no brow- nish black particles with the negative control in cells. Some blue granules of Tropic hematox- ylin can be seen in the nucleolus.4 The result of osteoblast BMP2 Immunohistochemistry: compared with the normal group, the expression level of Gushukang group and Kidney-tonifying group raises obviously (P < 0.01), tonifying spleen group,MG132+ normal group, MG132+ Gushukang group and MG132+ tonifying spleen group reduce significantly (P < 0.01). Compared with the Kidney- tonifying group, the expression of other groups reduce significantly (P < 0.01).5 The result of osteoblast BMP7 Immunohistochemistry: compared with the normal group, the expression level of Kidney-tonifying group raises obviously (P < 0.01), the expression level of Gushukang group, tonifying spleen group,MG132+ normal group, MG132+ Gushu- kang group, MG132+ Kidney-tonifying group and MG132+ tonifying spleen group reduce significantly (P < 0.01). Compared with the Kidney-tonifying group, the expression of other groups reduce significantly (P < 0.01). Compare each group with herbs serum, the expression of MG132+ normal group was the highest, the Kidney-tonifying group was little reduce.6 The result of osteoblast Smad1/5 Immunohistochemistry: compared with the normal group, the expression level of Gushukang group,Kidney-tonifying group, tonifying spleen group, MG132+ normal group, MG132+ Gushukang group, MG132+ Kidney-tonifying group and MG132+ tonifying spleen group reduce significantly (P < 0.01). Compared with the Kidney- tonifying group, the expression level of Gushukang group, tonifying spleen group,MG132+ normal group, MG132+ Gushukang group, MG132+ Kidney-tonifying group and MG132+ tonifying spleen group reduce significantly (P < 0.01). Compare each group with HERBS serum, the expression of MG132+ normal group was the highest. Compare each group with the Kidney-tonifying group, the expression level of MG132+ normal group raises obviously (P < 0.01), MG132+ Gushukang group and MG132+ tonifying spleen group reduce significa ntly (P < 0.01).Conclusion:1 The method to get rats' osteoblast with repeated collagenase digestion to cultivate it in vitro is reliable, simple and easy, which can provide a lot of highly puritied osteoblasts for scientif- ic research.2 There are positive expressions of BMP2,BMP7and Smad1/5 in the newborn rats' cranial cultivated in vitro, which means that there are BMP-Smad signal transduction pathways in normal rats' osteoblast.3 The kidney medicine serum can obviously promote the proliferation of osteoblast, increase significantly the expression levels of BMP2, BMP7, and Smad1/5 in osteoblast, which means that the HERBS of tonifying the kidney, marrow and bones can prevent and treat osteoporosis.4 Application of protease inhibitors of UPP pathways MG132 could inhibit the proliferation of bone cells, decrease the expression levels of BMP2, BMP7 and Smad1/5 in osteoblasts.5 Application of HERBS serum of tonifying the kidney, marrow and bones could inhibit the proliferation of osteoplasts by intervening the protease inhibitors MG132 and increase significantly the active expressions of BMP2, BMP7 and Smad1/5 in osteoblasts cultivated in vitro.