| Objective: Observing the variation of TGF-β1, Smad2/3 mRNA and protein expression in the hypothalamus of rats with osteoporosis due to kidney deficiency, exploring the molecular biology basement of osteoporosis due to kidney deficiency. researching the preventive and therapeutic effect mechanism of nanometer calcium kidney-tonifying herbs to the rats with osteoporosis due to kidney deficiency, Providing the experimental foundation to the secondary osteoporosis due to kidney deficiency which induced by glucocorticoid in clinical treatment.Methods: Build a rat model of secondary osteoporosis due to kidney deficiency by inject glucocorticoid in muscles the of posterior limb. Treat the rats with different dosage of nanometer calcium kidney-tonifying herbs for 8 weeks, while taking Gushukang Keli, Gaitianli as positive control groups and also set normal groups, model groups. Mensurate the bone density of the left femur of the rats with Model XR-36 double energy X-ray skeletal density detector. Detect the indexes of TRAP and ALP of the serum with biochemical methods. Detect the TGF-β1, Smad2/3 mRNA and protein expression in the hypothalamus of rats with kidney-deficiency osteoporosis by using RT-PCR and Western imprinting.Results:(1)BMD data of the left femur of rats: After 8 weeks, model gourp got an obviously decrease of the level of BMD (p＜0.01); and all the medicines groups (nanometer calcium kidney-tonifying herbs, Gushukang Keli and Gaitianli) have increased the level of BMD compare with the model groups, while the high dose groups has got the best effects(p＜0.01).(2)TRAP-biochemical index of serum: the model groups got an obviously increase of TRAP level compared with normal groups(p＜0.01); all the medicines were able to decrease TRAP level significantly compared with model groups(p＜0.01).(3)ALP-biochemical index of serum: The model groups (male and female) got an obviously increased the level of ALP compared with normal groups(p＜0.01); the ALP level is incresed in allthe groups with medicine(p＜0.01). the nanometer calcium kidney-tonifying herbs groups with the high dose (male and female), Gushukang Keli and Gaitianli groups have obviously increased the ALP level compared with model groups (p＜0.01).(4)TGF-β1 mRNA and protein expression in the hypothalamus of rats: the model groups got a obviously decreased in TGF-β1 mRNA and protein expression compared with normal groups (p＜0.01); compared with model groups, nanometer calcium groups(high and low dose) and the Gaitianli groups can increase the TGF-β1 mRNA and protein expression to differrent level, the nanometer calcium kidney-tonifying herbs groups with high dose get the highest increase(p＜0.01).(5)Smad2/3 mRNA and protein expression in the hypothalamus of rats: model groups got a obviously decreasd in Smad2/3 mRNA and protein expression compared with normal groups (p＜0.01). all medicine groups increasd Smad2/3 mRNA and protein expression in hypothalamus to differrent level compared with model groups, nanometer calcium kidney-tonifying herbs gruops with high dose get the highest increase(p＜0.01).Conclusion:(1)The model of osteoporosis in rats due to kidney deficiency is able to be build successfully by intramuscular inject glucocorticoid.(2)Nanometer calcium kidney-tonifying herbs can distinctly increase the bone density, promote the formation of the bone and restrain the bone absorption.(3)Expresion the TGF-β1 and its signal transduction element Smad2/3 on gene and protein level in the hypothalamus tissue in normal rats, It suggested there have signal transduction passageways of TGF-β1 and Smad2/3 in hypothalamus.(4)The decreasing of TGF-β1, Smad2/3mRNA and protein expression in the hypothalamus of rats maybe the one of the mechanism of osteoporosis due to kidney deficiency.(5)The level of TGF-β1, Smad2/3mRNA and protein expression in the hypothalamus of rats obviously increased by nanometer calcium kidney-tonifying herbs, which indicatedthe chinese herbs with kidney-tonifying, calcium-supplying action are able to prevent and cure osteoporosis due to kidney deficiency indirectly by inducing the expression of TGF-β1 and Smad2/3 in the hypothalamus. |
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