Kidney In Rats With Osteoporosis And Osteoblasts Of Tgf-beta <sub> 1 </ Sub> And Its Signal Transduction Molecules Of Smad2, 4 Gene Regulation Mechanism

Purpose: In order to explore the etiological factors, pathology and the preventing and treating mechanism of the osteoporosis due to kidney deficiency, we researched the regulating mechanism of TGF-β1 and its signal transduction gene and protein - Smads in the rats of osteoporosis due to kidney deficiency and in vitro cultured osteoblast from the rat calvarias. Materials and Method: The experiment includes two parts:in vivo part and in vitro part. In vivo part, we established the rat model of osteoporosis due to kidney-deficiency, using the method of ovariectomy, treating the osteoporosis rat with MIGUKELI (big dose, middle dose, small dose group) , the function of MIGUKELI is tonifying Qi and strengthening essence, invigorating the kidney to make the bone strong. Simultaneously, using GUSHUKANG and nilestriol to be the control group for 12 weeks. And rats in the normal group and model group were not given any treatment. We had used dual energy X-ray bone density instrument to detect the bone mineral density of the rats; detected the Alkaline phosphatase, tartrateresistant acid phosphatase, osteocalcin, estrogen, calcium, magnesium, phosphorum in the blood serum and the calcium, creatinine , hydroxyproline in the urine with the method of biochemistry and radio-immunity; observed the morphometry of the bone tissues with the method of HE stain; detected the activity of TGF-β1 in the blood serum of the rats with the method of ELISA; We had used the immunohistochemistry, RT-PCR, western blotting method to investigate the expression of TGF-β1 mRNA and Smad2,4 mRNA and proteins. In vitro part, we cultured the osteoblast of the newborn rats calvarias; identified the osteoblast cultured in vitro with the method of enzyme cytochemistry (ALP-Gomori) and evaluated the proliferation of the osteoblast after the action of serum contained tonic-kidney herbs; detected the expression of TGF-β1 and Smad2, 4 proteins in the osteoblast with the method of immunohistochemistry;the expression of mRNA of TGF-β1 and Smad2, 4 gene were detected by RT-PCR.Result: 1. In vivo part: (1) The comparison of uterus exponent of the experimental animals: the uterus exponent of the model groupis lower than that of the normal group evidently, the MIGUKELI groups and GUSHUKANG group can increase the uterus exponent only in some extent but have no significance in statistics. The nilestriol group can increase the uterus exponent significantly in statistics. (2) The comparison of the morphometry of the bone tissues in the experimental rats: we observed the HE stain spongy bone parts of the proximal of the rats right tibia in the LM: in normal group, the shape and structure of the bone trabecula are complete, and the arrangement are normal; In the model group, the arrangement of bone trabecula structure is rarefaction, it appeared many caecum of the bone trabecula, the bone trabecula thickness ( Tb. Th). decreased, the marrow cavity area rates (%Ma. Ar) increased obviously; while in the MIGUKELI groups and GUSHUKANG group, nilestriol group, The amount of the bone trabecula increased, the bone trabecula became more widely, the arrangement of them changed to more tightly, Tb. and Th. increased and %Ma. Ar decreased. (3) The comparison of the blood and urine biochemistry index of the experimental rats: the contents of Ca, P, Mg in the blood serum and the Ca/Cr in the urine of all the groups have no significance in statistics. Comparing to the normal group, the activity of BGP, TRAP, ALP in the blood serum and the ratio of Hop/Cr in the urine increased evidently, the level of E2 in the blood serum decreased remarkably; Comparing with the model group, the activity of BGP, ALP in the blood serum of the MIGUKELI groups, GUSHUKANG group and nilestriol group decreased remarkably, the activity of TRAP also decreased but it didn' t achieve the level of the normal group. And the ratio of the Hop/Cr decreased obviously. (4)The comparison of the BMD of the femur and tibia of the experimental rats: the BMD of the femur and the proximal part of the femur,...