| Objective: qRT-PCR was used to detect the expression of miRNA-708 in BMSCs,osteoblasts and adipocytes of steroid induced osteonecrosis of femoral head in rabbits.Methods: 50 New Zealand white rabbits were divided into blank group(group A)and experimental group(Group C).The experimental group(40 rabbits)was made of micro LPS combined with methylprednisolone sodium succinate.Four weeks later,8rabbits died and the remaining 32 rabbits were randomly selected for MRI detection.After the success of modeling,10 rabbits were divided into model group(group B),11 rabbits in Xianlinggubao group(Group C)and 11 rabbits in Shengguzaizao pill group(Group D).According to the pre experimental design,group C was treated with0.4g/kg Xianlinggubao,group D with 0.6g/kg shengguzaizao,group A and group B with the same volume of normal saline.During the period of gastric perfusion,the general situation of New Zealand white rabbits in each group was closely observed.Four weeks later,both sides of the femur were taken out under sterile condition by air embolism.The right femoral head was sectioned for pathology.After he staining,the empty bone lacuna and pathological changes of each group were observed.BMSCs,osteoblasts and adipocytes were extracted from the left femur.The expression differences of miRNA-708-3p and miRNA-708-3p in different cells of each group were analyzed by qRT-PCR.Results:(1)General situation of New Zealand white rabbits: before modeling,each group of New Zealand white rabbits had good mental condition,glossy hair,no abnormal smell,normal diet,normal movement of limbs and joints,and sensitive response.After the establishment of the model,the New Zealand white rabbits in the experimental group were depressed in spirit,decreased in activity,decreased in diet and drinking water,and were slow in response.Some of the experimental rabbits had muscle strength and lameness in their hind limbs.Compared with group B,the mental state,diet,muscle strength of hind limbs and lameness of group C and group D were improved,but they did not return to normal compared with group A.(2)MRI findings: in the experimental group,MRI showed that the articular surface of the femoral head of New Zealand white rabbits was complete,the femoral head did not deform,the joint space was normal,the load area of the upper front part of the femoral head showed abnormal signal in flake,and the low signal T2W2 showed high signal on T1 WI.(3)He staining: there was no obvious abnormal pathological change in group A,the local bone trabeculae and related cells in the experimental group had abnormal pathological changes in varying degrees,empty bone lacunae increased significantly.The rate of empty bone lacuna in group B was significantly higher than that in group A(P<0.001);the rate of empty bone lacuna in group C and group D was significantly lower than that in group B(P<0.001),the difference was statistically significant;the rate of empty bone lacuna in group C and group D was not statistically significant(P>0.05).(4)qRT-PCR showed that the expression level of miRNA-708-3p in group B was higher than that in group A(P<0.01),The expression level in group C and D was lower than that in group B(P< 0.01),There was no significant difference between group C and group D(P>0.05).The expression level of miRNA-708-5p in group B was higher than that in group A(P<0.01),in group C and group D was lower than that in group B(P<0.01),and in group D was lower than that in group C(P>0.05).The expression level of miRNA-708-3p in osteoblasts was higher in group B than in group A(P<0.01),and lower in group C and D than in group B(P<0.01),There was no significant difference between group C and group D(P>0.05).The expression level of miRNA-708-5p in group B was higher than that in group A(P<0.01),in group C and group D was lower than that in group B(P<0.01),and in group D was lower than that in group C(P>0.05).The expression level of miRNA-708-3p in group B was higher than that in group A(P<0.01),There was no significant difference between group C and group D(P>0.05).The expression level of miRNA-708-5p in group B was higher than that in group A(P<0.01),There was no significant difference between group C and group D(P>0.05).Conclusion:(1)SANFH can be successfully reproduced by the method of microLPS combined with methylprednisolone sodium succinate.(2)The overexpression of miRNA-708 in MSCs and osteoblasts may be one of the main mechanisms of steroid induced necrosis of the femoral head.(3)It may be one of the main mechanisms of the treatment of SANFH that shengguzaibao pills target to regulate the expression level of miRNA-708 in BMSCs and osteoblasts.(4)Shengguzaizao pill is more effective than Xianlinggubao in regulating miRNA-708-5p. |
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