| Objective:To detect effects of boldine in the collagen induced arthritis (CIA) rat model and the possible mechanism of action.Methods:The CIA rat model was established in all rats, except those n the blank control group using bovine type II collagen and incomplete Freund’s adjuvant. Then the rats were subdivided into the following groups:(1) blank control group; (2) CIA model; (3) MTX group (3mg/kg, twice/week); (4) the low dose of boldine group (2mg/kg/d); (5) the high lose of boldine group (4mg/kg/d). Drug treatment was given orally beginning 15 days after the first immunization for 28 days. During the period of the drug treatment, the general situation of rats was observed, at body weight measured, AI changes judged and ankle swelling degree neasured with the vernier caliper. After the end of treatment, the expression levels of the inflammatory cytokines in serum by enzyme linked immunosorbent method such as IL-10 was performed in CIA rats.Meanwhile, HE staining was executed to observe the histopathology changes of the ankle and immunohistochemical staining method carried out to detect the expression level of IL-10 and IL-17 in local rat ankle oint. As well as indices of thymus and spleen was observed. All of these vere completed to evaluate the effect of boldine and explore their nechanism. Then take the vertebrae (lumbar) and femur for the detection on BMD and micro-CT.Results:1 Evaluation arthritis index and general state:Compared with blank control group, rats of model group had significantly weight loss, activities reduced and spirit was poor. Progression of arthritis symptom after building especially as the symmetric hind limbs joint significantly. The peak of arthritis performance appears in 4 weeks after the immune, then gradually into the chronic phase with joint stiffness and deformity.2 Ankle joint swelling and histopathological results showed:the high dose of boldine group could significantly alleviate the degree of inflammation in model rats. During treatment, on day 10, AI values of high dose of boldine group was lower than that of the CIA group (P <0.05); From the beginning of the 18th day, AI value of each drug group was significantly lower than that of the CIA group (P<0.05). After 10 days, the foot plantar thickness in high dose of boldine group was lower than that in the CIA group (P<0.05); on day 28, the foot plantar thickness in the high dose of boldine was significantly decreased. By pathology observation and analysis, in addition to the MTX group, other drug groups had different degree relief in pathological changes of the ankle joint, especially the high dose of boldine group.3 Cytokine levels according to the results showed:Compared with the blank control group, IL-10 serum levels in CIA group had significantly decreased (P<0.01). Other drug group had different increased in IL-10 serum, especially the high dose of boldine group.4 BMD on lumbar to the results showed:Compared with blank control group, BMD of CIA group was obviously lower (P< 0.01); Compared with CIA group, the high dose of boldine group was significantly higher on BMD(P< 0.01).5 Micro-CT of proximal femur to the results showed:Compared with the blank control group, bone volume/total volume (BV/TV), trabecular number (Tb.N) and trabecular thickness (Tb.Th) in CIA group was significantly decreased(P< 0.05 or P< 0.01), while trabecular pattern factor (Tb.Sp), bone surface area/bone volume(BS/BV) and trabecular pattern factor (Tb.Pf) was distincted increased(P< 0.05 or P< 0.01). Compared with the CIA group, BV/TV, Tb.N and Tb.Th in MTX group was increased(P< 0.05 or P < 0.01), and Tb.Sp, BS/BV and Tb.Pf had lower decreased(P< 0.05 or P< 0.01). Boldine groups had different increased in BV/TV、Tb.N、 Tb.Th (P< 0.05 or P< 0.01), while Tb.Sp、BS/BV、Tb.Pf were distinctly decreased (P< 0.05 or P< 0.01).Conclusions:1 Type II collagen and incomplete Freund’s adjuvant immunized animal model induced animal models of rheumatoid arthritis, and could appear lumbar and proximal femoral bone micro structure change of state. Boldine may reduce the degree of joint swelling of CIA rats, relieved the pathological injury of the joint, and could significantly alleviate microstructure degeneration and bone loss in CIA rats. Boldine with high dose (4mg/kg) treatment group show excellent effection, it indicated that Boldine maybe have the potential preventive role for rheumatoid arthritis secondary systemic osteoporosis.2 Boldine could increase the level of IL-10 and down-regulation of IL-17 levels, relieve joint synovitis lesions, which may be one of the mechanisms it treated CIA model. |
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