Experimental Study Of Lovastatin And Batroxobin On Treatment Of Collagen-induced Arthritis In Rats

PartⅠTreatment and Immunological Mechanism of the Protective Effect of Lovastatin on Collagen-induced Arthritis in RatsObjective:To evaluate the efficacy and immunological mechanism of lovastatin on the treatment of collagen-induced arthritis(CIA) in rats and to supply theoretical basis for exploring new treatments of inflammatory arthritis.Methods:Arthritis of rat was induced injecting bovineⅡcollagen(BIIC) to Wistar rats.The rats were randomly divided into five groups:normal control group,CIA control group treated with normal saline,ibuprofen group,Methotrexate(MTX) group and Lovastatin group.The body weight,the hind paw volume,the arthritic index of all rats at different time points were observed and measured.The therapeutic effects of different reagents were assessed by measuring the change of serum cytokine levels including TNF-α,IL-6,IL-8 and IP-10 level..At the same time,the expression of IL-1βand IL-17mRNA'were detected by QRT-PCR.At the end of experiment,the radiographic changes and the synovial pathology score of rat ankle joints were evaluated.The expressions of MCP-1 and RANTES in the synovium of ankle joint were evaluated and the pathogenic roles of MCP-1 and RANTES in CIA in rats were analysed.Results:1.The model of CIA was constituted successfully at 11～13 days after first immunization of typeⅡcollagen.At the 17^th day,the swelling of dorsum of hind feet and ankle joints deteriorated.The sole of the rats were thick.The mobility and appetite of the rats decreased.2.The degree of swelling of ankle joints in Lovastatin group was decreased since the 3^th week.At week 4,the volume of hind paw in Lovastatin group was lower than that of CIA group.At the 5^th week,the arthritis index of Lovastatin groups was lower than that of the CIA group.3.The serum levels of TNF-α,IL-6 and IP-10 in Lovastatin group were decreased since 5^th week.The expressions of IL-1βand IL-17mRNA' in Lovastatin group were less than that of the CIA group.4.Radiologic finding:The soft tissue around the ankle joint was swollen. Osteoporosis and and bone erosion were not found in Lovastatin group.5.Histopathologic results:Comparing with CIA group,the degree of synovial swelling in Lovastatin group was decreased.The synovial proliferation and inflammatory cell infiltration were mild.The vascular hyperplasia and pannus were significantly declined.6.At the 5^th week,the cells expressing MCP-1 and RANTES were found in the interface layer of synovium and cartilage in CIA group.Few positive cells of MCP-1 and RANTES were found in Lovastatin group.Conclusion:1.Lovastatin significantly ameliorates the symptoms of CIA rats,and inhibits the development of inflammatory swelling.2.Lovastatin can decrease the expression of TNF-α,IL-1β,IL-6,IP-10, IL-17,MCP-1 and RANTES.It indicates Lovastatin may play a protective role on joint erosion through regulating the expression of these cytokine.3.MCP-1 and RANTES have important roles in CIA rats,because they expressed on CIA rats' joints. PartⅡTreatment and Immunological Mechanism of the Protective Effect of Batroxobin on Collagen-induced Arthritis in RatsObjective:To evaluate the efficacy and Immunological Mechanism of Batroxobin on the treatment of CIA in rats and to supply theoretical basis for exploring new treatments of inflammatory arthritis.Methods:The CIA rats were randomly divided into different groups:normal control group,CIA group treated with normal saline,Ibuprofen group, methotrexate(MTX) group,high dose and low dose Batroxobin groups.Several indexes(weight,volume of hind paw,arthritic index,radiography of ankle joint) were observed in different groups.The plasma fibrinogen level in each group was detected.The levels of TNF-α,IL-6,IL-8,IP-10,MCP-1 and RANTES in each group were measured by ELISA.The expressions of IL-1βand IL-17mRNA were detected by QRT-PCR.The histopathology of synovium was scored.Results:1.The model of collagen-induced arthritis was constituted successfully at 11～13 days after first immunization of typeⅡcollagen.At the 17^th day,the swelling of dorsum of hind feet and ankle joints deteriorated.The sole of the rat was thick.The mobility and appetite of the rats decreased.2.The degrees of swelling of ankle joints in high dose and low dose of Batroxobin groups were decreased since the 3^th week.At week five,the volume of hind paw in high dose Batroxobin group was lower than that of the CIA group.At the 5^th week,the arthritis indexes in both Batroxobin groups were lower than CIA group.3.The plasma fibrinogen level of in each group wae detected at 2^th and 5^th week.The plasma fibrinogen level in high dose of Batroxobin group was lower than that of the CIA group at 2^th week.And the plasma fibrinogen levels in both high and low dose Batroxobin groups were lower than that of the CIA group at 5^th week.4.The levels of TNF-α,IL-6 and RANTES in both high and low dose Batroxobin groups were decreased since 5^th week.The levels of IP-10,IL-8 and MCP-1 in high dose of Batroxobin were lower than the CIA group.The expressions of IL-17mRNA's in both Batroxobin groups were less than that of the CIA group.The expressions of IL-1βmRNA in both Batroxobin groups were decrease.But there were no stastistically difference when compared with CIA control group at 5^th week.5.Radiologic finding:There was soft tissue swelling around the ankle joint but osteoporosis and bone erosion were not found in Batroxobin group.6.Histopathologic results:Comparing with CIA group,the degree of synovial swelling in Batroxobin group was decreased.The synovial proliferation and inflammatory cell infiltration were mild.The vascular hyperplasia and pannus were significantly declined.Concluion:1.Batroxobin significantly ameliorates the symptoms of CIA rats,and inhibits the development of inflammatory swelling.2.Batroxobin can decrease the plasma fibrinogen level and inhibit the actue reaction protein.3.Batroxobin can decrease the expression of TNF-α,IL-1β,IL-6,IP-10, IL-17,MCP-1 and RANTES.Indicates Batroxobin may play a protective role on joint erosion through regulating the expression of these cytokine.