| Objective By comparing internal heat-type acupuncture needle on the behavior,subchondral bone morphology and subchondral osteoprotegerin(OPG),receptor activator of NF-kB ligand(RANKL)and Changes in the expression of receptor activator of NF-kB(RANK),to explore the curative effect and partial mechanism of internal heat acupuncture therapy on KOA.Methods 1.Grouping and model preparation: Using a random number table method,30 SPF healthy New Zealand rabbits were divided into a blank group(10)and a model group(20).The improved Hulth modeling method was used to prepare the rabbit left hind limb KOA model,and after the modeling was successful,it was randomly divided into a model group(10animals)and a treatment group(10 animals);2.Intervention: The experimental rabbits in the treatment group received internal thermal acupuncture treatment(42℃,1 time/week,20 minutes each time)on the left posterior knee joint of the experimental rabbits for 4consecutive weeks;neither the model group nor the blank group were given any treatment;3.Observation Indexes: The Lequesne MG scale was used to score the gait of each group of experimental rabbits.Toluidine blue staining(TBS)was used to observe the morphological changes of the subchondral bone in each group of experimental rabbit knee joints;Western blot and RT-PCR were used to detect the levels of OPG,RANKL,RANK protein and their mRNA in the subchondral bone expression.Results 1.Lequesne MG gait score Before intervention,compared with the blank group,the experimental rabbit behavior scores of the model group and the treatment group were significantly increased(P<0.05);the difference between the treatment group and the model group was not statistically significant(P>0.05).After intervention,the difference between the treatment group and the model group was statistically significant(P<0.05).The difference between the treatment group after intervention and before intervention was statistically significant(P<0.05).2.Toluidine blue staining Compared with the blank group,the trabecular bone of the model group showed a sparse distribution,with interrupted continuity,decreased trabecular bone content,and increased spacing.Compared with the model group,the trabecular bone density of the treatment group increased,the distribution was even,and the spacing became narrower.3.Comparison of bone tissue morphometric parameters Compared with the blank group,the number of trabecular bones in the model group decreased,the percentage of trabecular bone area decreased significantly,and the trabecular bone separation became larger(P<0.05).Compared with the model group,the separation of trabecular bone in the treatment group decreased significantly,and the number and area percentage of trabecular bone increased(P<0.05).The trabecular bone width of the three groups did not change significantly,and there was no statistical difference(P=0.549).4.Expression of OPG,RANKL and RANK proteins in subchondral bone tissues Compared with the blank group,the expression of OPG in the model group was significantly reduced,and the expression of RANKL and RANK was increased(P<0.01);compared with the model group,the expression of OPG in the treatment group was significantly increased,and the expression of RANKL and RANK was decreased(P< 0.05).5.Expressions of OPG,RANKL and RANK mRNA in subchondral bone tissues Compared with the blank group,the expression of OPG mRNA in the model group was significantly reduced(P<0.05),and the expression of RANKL and RANK mRNA increased(P<0.01).Compared with the model group,OPG mRNA expression in the treatment group increased significantly,and RANKL and RANK mRNA expression decreased(P<0.05).Conclusion Internal heat-type acupuncture needle can significantly improve the behavior of rabbit KOA and subchondral bone morphology.The mechanism may be achieved by up-regulating the expression of OPG in rabbit KOA subchondral bone and down-regulating the expression of RANKL and RANK. |
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