| 1 Objective:With the mass use of clinical application of hormone, the trend of SONFH is increasing. SONFH is a difficult and complicate disease as its pathogenesis is not clear and lack of certain effective treatment approach for SONFH. Traditional Chinese medicine shows superiority in terms of prevention and treatment. But to the advanced SONFH, it's unsatisfactory when simply uses traditional Chinese medicine. To study the therapeutic effects and mechanism of the prescription includes supplement kidney, invigorate blood and dispel phlegm and prescription of Jian Gu on rabbit's SONFH prevention.2 Methods30 healthy SPF New Zealand rabbits were randomly allotted to 4 groups by weight:normal group (6 rabbits), the model group (8 rabbits), the prescription includes supplement kidney, invigorate blood and dispel phlegm and prescription of Jian Gu group (8 rabbits High-dosage group group for short), and the prescription of Jian Gu group (8 rabbits Low-dosage group group for short).The normal control group was intragastrically perfused with saline of the same volume as 10 ml per kilogram at the same time. Low-dosage group were intragastrically perfused with suspension of prescription of Jian Gu with same volume (effective components is 3.6g/kg/d); and High-dosage group is 7.19g/kg/d. After gavage for 3 weeks, the model group, Low-dosage group and High-dosage group were injected sodium succinate methylprednisolone on the internal side of left leg 20mg/kg each time for another 3weeks. After the experiment for 3 weeks, physical examination such as TC, TG, HDL and hemorrheological changes were collected for detection. Part of the centrifugal blood kept on the -70℃would be test for OPG and RANKL. After injected by sodium succinate methylprednisolone for 3 weeks, TC, TG, HDL and hemorrheological changes were be measured again. All the blood samples were determined for OPG and RANKL by enzyme linked immunosorbent assay (ELISA) especially include the sample before modeling; and killed the rabbits, took the right posterior femoral head, which fixed formalin then observed pathologically with frost slice techniques and Sudan staining. It was observed under light microscopy from lower power to higher power according to Matsui grades to measure if the marrow and bone trabeculae were necorsis.Date was tested with Image-Pro Plus 6.0 software to calculate the rate and square of necrosis of femoral head. The rest of the date was tested with SPSS18.0 software. The results were measured by ((?)±S). mean comparison in groups was conducted with single factor variance analysis, and the pairwise comparison was performed with S-N-K method. Correlation between the variable was deal with Spearman. There is statistical significance when P<0.05, and a very significant difference when P<0.01.3 Results3.1 The general result:One rabbit was died as faulty intragastrically method to block the tracheobronchial and made acute asphyxia in the Low-dosage group group. And 2 rabbits were died after modling.3.2 Compare between blood lipids level:HDL,TCHO and TG blood level before the experiment is F=0.733, P=0.542>0.05;there is no statistical significance in the pairwise comparison with S-N-K method (P>0.05), which shows comparable. After the experiment:①compare between each group in the level of HDL:F=2.534, P=0.082>0.05; the pairwise comparison with S-N-K method shows that:the HDL blood level in the model group was lower than normal group and had a statistical significance (P=0.042<0.05); the HDL blood level in the model group was lower than High-dosage group and had a statistical significance (P=0.018<0.05).②compare between each group in the level of TCHO, F=0.458, P=0.714>0.05; there is no statistical significance in the pairwise comparison with S-N-K method (P>0.05).③compare between each group in the level of TRIG:F=5.041, P=0.008<0.01; the pairwise comparison with S-N-K method shows that:the TRIG blood level in the model group was higher than normal group and had a very statistical significance (P=0.001<0.01); higher than low-dosage group (P=0.019<0.05) and high-dosage group (P=0.013 <0.05).3.3 Compare between hemorrheological changes:index of hemorrheological changes before the experiment is P>0.05, there is no statistical significance in the pairwise comparison with S-N-K methodwhich shows comparable. After the experiment:①compare between each group in the level of whole blood viscosity: there was no statistical significance(P>0.05) between low-dosage group and model group;model group was much higher than normal group (P<0.01); blood viscosity 200 of model group was higher than High-dosage group (P<0.05); blood viscosity. (30,5,1) of model group was much higher than High-dosage (P<0.01);blood viscosity 200 of Low-dosage group was higher than normal group (P<0.05);blood viscosity (30,5,1) of Low-dosage group was much higher than normal group (P<0.01).②compare between each group in the level of TCHO: F=0.458, P=0.714>0.05; there is no statistical significance in the pairwise comparison with S-N-K method (P>0.05).③compare between each group in the level of TRIG:F=5.041, P=0.008<0.01; the pairwise comparison with S-N-K method shows that:the TRIG blood level in the model group was higher than normal group and had a very statistical significance (P=0.001<0.01); and higher than low-dosage group (P=0.019<0.05) and high-dosage group (P=0.013<0.05); The low-dosage group was lower than normal group in hemorrheological changes, there was a very statistical significance between two groups.3.4 Compare between OPG,RANKL,OPG/RANKL:OPG,RANKL,OPG/RANKL blood level before the experiment is F=0.046, P>0.05; there is no statistical significance in the pairwise comparison with S-N-K method (P> 0.05), which shows comparable. After the experiment:①compare between each group in the level of OPG:F=6.667, P=0.002<0.01; the pairwise comparison with S-N-K method shows that:the OPG blood level in the model group was much lower than normal group and had a very statistical significance (P=0.000<0.01); was lower than Low-dosage group and had a statistical significance (P=0.026<0.05); and was lower than High-dosage group and had a statistical significance (P=0.034< 0.05); the OPG blood level in Low-dosage group and High-dosage group was lower than normal group and had a statistical significance (P<0.05).②compare between each group in the level of RANKL:F=0.616, P=0.612>0.05; there is no statistical significance in the pairwise comparison with S-N-K method (P> 0.05), which shows comparable.③ompare between each group in the level of OPG/RANKL:F=14.146, P=0.000<0.01;the pairwise comparison with S-N-K method shows that:the OPG/RANKL in the model group was much lower than normal group,Low-dosage group and High-dosage group and had a very statistical significance (P<0.01); the OPG/RANKL level in Low-dosage group and High-dosage group was lower than normal group and had a very statistical significance (P<0.01)3.5 Pathology of femoral head:①ormal group:all of the 6 femoral head pathology were normal with 0 grade pathology;model group:2 rabbits were died after modling, the other 6 femoral head pathology were 2 grade pathology;Low-dosage group:1 femoral head pathology was 2 grade pathology and 6 femoral head pathology were 1 grade; pathology high-dosage group:1 femoral head pathology was 2 grade pathology and 7 femoral head pathology were 1 grade.②the necrotic area and necrotic rate of normal group were much higher than the other three groups and had a very statistical significance (P<0.01).3.6 Correlation analysis between OPG,RANKL,OPG/RANKL in blood and pathology of femoral head:there is negative correlation between OPG and necrotic area (R=-0.638, P=0.00<0.01) and has much statistical significance; there is no correlation between RANKL and necrotic area (R=0.131, P=0.516>0.05) and has no statistical significance;there is negative correlation between OPG/RANKL and necrotic area (R=-0.636, P=0.00< 0.01) and has much statistical significance;there is negative correlation between OPG and necrotic rate (R=-0.673, P=0.00<0.01) and has much statistical significance; there is no correlation between RANKL and necrotic rate (R=0.142, P=0.479>0.05) and has no statistical significance; there is negative correlation between OPG/RANKL and necrotic rate (R=-0.638, P=0.000< 0.01) and has much statistical significance.4 Conclusion4.1 Model of SONFH by injected sodium succinate methylprednisolone on the rabbit is an easy way. Because it could shorter the production cycle and decrease the mortality of the rabbit.4.2 Hormone would cause abnormal blood lipid metabolisom and blood rheology, which lead to decrease the level of OPG. OPG/RANKL in rabbit. Also it could increase the activity of osteoclasts and bring down the activity of the osteoblast-like cells so that cause SONFH happen.4.3 Prescription includes supplement kidney, invigorate blood and dispel phlegm and prescription of Jian Gu can prevent rabbit's SONFH happened.4.4 In the experiment, both the high-dosage group and low-dosage group can obviously improve the abnormal blood lipid, hemorrheological changes and blood viscosity. And the hight dosage, the more effect in the experiment. Maybe it would prevent SONFH through improving the abnormal blood lipid, hemorrheological changes cause by hormone.4.5 Prescription includes supplement kidney, invigorate blood and dispel phlegm and prescription of Jian Gu can improve the OPG, low level of OPG/RANKL in the blood which was caused by hormone. It could help to bring down the activity of osteoclast and increase the osteoblast in order to strength the density of the bone and prevent the bone collapsed and degraded. |
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