| Objective: The effect of combination with Guizhi and Fuzi transfer into the blood and their interaction under the environment on signaling pathway of OPG/RANKL/RANK was observed,to explore the mechanism of bone protection for RA,and confirmed the hypothesis: Combination with Guizhi and Fuzi environment plays a role in bone protection by regulating signaling pathway of OPG/RANKL/RANK to resist RA bone erosion.To improve the clinical effect of traditional medicine in RA prevention and treatment,this study explored a method for prevention and treatment of RA bone erosion by Chinese medicineMethods: The rat model of the CIA was prepared by type II collagen,the serum was taken from the stomach,and the expression of OPG,RANK and RANKL of osteoclast were detected by enzyme-associated immunosorbent assay(ELISA).Fifty Wistar rats were randomly divided into normal group(n=10)and CIA model(n=40)made by bovine collagen ⅱ.According to the inductive reference of collagen arthritis in Wistar rats,100 mg bovine type Ⅱ collagen was dissolved in 10 mL 0.1M acetic acid,then mixed with the same amount of complete Freund’s adjuvant at 4℃ for the night.The final concentration of type Ⅱ collagen was 5mg/mL.Then the syringe was pumped repeatedly,until the mixture was completely emulsified so that the emulsified matter was not loosened into the water.The emulsified mixture was injected into the caudal root of rats with 0.1 m L of 0.1mL in the back of the skin,each of which was given 1 mg of collagen Ⅱ,and the injection site was compressed for 30 s,so that the emulsified substance could be absorbed completely.The normal control group was given the same dose of normal saline and 0.1mL wasinjected into the caudal root of rats.Day 14 boosts immunization once.The other 10 rats were divided into three groups: low dose group,middle dose group and high dose group,with 10 rats in each group.The low,medium and high dosage of medicine was given by stomach in the control group and the model group respectively,and the volume of the drug was 1ml/100 g in the normal group and the model group,and the volume of the drug was1ml/100 g in the normal group and the model group,and the volume of the drug was 1ml/100 g.Once a day for 4 weeks.The rats were anesthetized with0.3% pentobarbital sodium 30mg/kg.The abdominal cavity was cut open.The abdominal aorta was exposed and ligated with surgical instruments.Take 5ml of arterial blood and centrifuge 10 min at 2000r/min on centrifuge,then extract supernatant.By means of ELISA analysis,contents of signaling pathway of OPG/RANKL/RANK in serum of rats were determined by comparing the drug containing serum with the normal serum extracted under the same conditions and the serum of the model group,and the results were compared with those of the control group and the model group.To investigate the interaction of OPG/RANKL/RANK signaling pathway in the treatment of rheumatoid arthritis(RA)with the combination of Aconiti Cinnamomi and Aconitum sinensis,and to explore the mechanism of bone protection in the treatment of rheumatoid a rthritis.Results:(1)The body weight of the mice increased significantly after feeding with feed before model making.After injection of collagen type II,the mice in the model group and the administration group could lose weight,but there was no significant difference between the two groups.(2)Compared with the normal group,the expression of OPG in the serum of the model group was significantly decreased by ELISA assay.The expressi on of RANKL and RANK was significantly increased in the model group.while the expression level of RANK and RANKL decreased(P<0.05).(3)Compared with the model group,the expression of OPG increased in each group(P<0.05),while the expression of RANK and RANKL decreased relatively(P<0.05).The expression of OPG in the medium and high groups was significantly higher than that in the low dose group,and the expression of OPG increasedwith the increase of the dose,while the expression of RANK and RANKL decreased.Conclusion:(1)The study showed that the weight of rats increased slowly compared with the normal group,because tdeterioration of bone erosion after modeling,which affected the normal growth and development of rats.(2)Combination with Guizhi and Fuzi can promote the secretion of OPG competitive binding RANKL,reduce the combination of RANK and RANKL,inhibit osteoclast differentiation,inhibit the bone erosion.Combination with Guizhi and Fuzi test in the high dose of low impact OPG/RANKL/RANK signal pathway,and with the increasing dose of aconite Guizhi effect is more obvious,so combination with Guizhi and Fuzi transfer into the blood and their interaction under the environment aconite RA bone protection mechanism,can increase the expression of OPG is regulated by OPG/RANKL/RANK signaling pathway,the expression of RANK and RANKL declined to achieve. |
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