Effects of BiZhongXiao Decoction on Annexin1 and Aldolase A Expression in Synovium of Collagen-induced Arthritis Rat
|School||Central South University|
|Course||Traditional Chinese Medicine|
|Keywords||collagen-induced arthritis synovitis rheumatoid arthritis Chinese Metrica annexin 1 aldolase A Western blotting|
Objective:(1) To elucidate the mechanism of the pathogenesis of rheumatoidarthritis(RA) by exploring the expression of annexin 1 and aldolase A insynovium of collagen-induced arthritis(CIA) rat;(2) To elucidate the mechanism of bizhongxiao decoction (BZXD)in treating RA by analyzing the effects of BZXD on annexin 1 andaldolase A expression in synovium of CIA rat.Methods:(1) The experiment arthritis rat model was established by injectedsubcutaneous of bovineⅡcollagen(BⅡC) and complete Freunds adjuv-ant(CFA).The CIA rats were divided into 2 groups randomly, the Bizhong-xiao decoction treatment group(BZXD group, n=19) drenched withBZXD and the CIA model group(model group,n=17), drenched with thesame dose of distilled water.Normal control group(Normal group, n=15)drank water freely.(2) General condition, weight, arthritis index score, degree of pawswelling and pathology was observed in 3 groups. The arthritis index(AI)of the rats was examined in CIA group and BZXD group on 10th day,15th day, 20th day, 25th day, 30th day, 35th day, 40th day and 45th dayrespectively. The weight and ankle thickness of rats was measured inthree groups every week. The pathomorphology of ankle was observed on25th day, 35th day,45th day.(3) Annexin 1 and aldolase A expression of synovium at differentstages from the normal, model and BZXD group were tested by Westernblotting, and analyzed by semi-quantification of gray scale with Imagetool 3.0 software. The correlation of annexin 1 and aldolase Abetween CIA group and BZXD group will be analysed.Results:1. Copying CIA model: CIA model could be copied by BⅡC andCFA with the incidence of 88.2% approximately.(1) General condition: poor appetite was shown in rats of modelgroup, more weight was losen than BZXD group(P＜0.05) as a result.Laterly, symptoms as dry and withering hair was manifested, whereassuch symptoms were not found in BZXD group. The weight increment inmodel group was lower than that in normal group in the 3rd w, the 4th w,the 5th w, the 6th w(P＜0.05),respectively, without significant differencebetween them in the 1st w, the 2nd w.The weight increment in BZXDgroup was higher than that in model group in the 5th w, the 6th w, withsignificant difference between them (P＜0.05), no significant differenceamong them at other times.(2) Arthritis index score evaluation: Arthritis of CIA rats was foundon 8-10 days after injection.Arthritis index score was found increased inmodel group and BZXD group on 10 days, reaching a peak on 25 daysand turning decreasing on 30 days in model group and BZXD group.But there was no manifest difference in arthritis index score betweenmodel group and BZXD group before 20 days(P＞0.05). 2 weeks aftertreatment, arthritis index score in BZXD group was decreasing. AI inBZXD group was lower than that in model group on 20d,25d,30d,35d,40d, 45d(P＜0.05),respectively.(3) Swelling degrees evaluation in joints of claws and feet: Theankle thickness of foot in model group was thicker than that in normalgroup in the 2nd w, the 3rd w, the 4th w, the 5th w, the 6th w (P＜0.05) respectively. The ankle thickness of foot in BZXD group was thinner thanthat in model group in the 5th w, the 6th w (P＜0.05,but no significantdifference among them at the other times.(4) Observation of pathology: Rat’s synovial tissues was not mani-fested by obvious newborn blood vessel and infiltrating inflammatorycells in normal group.While CIA model group on 25 days was manifestedby angiogenesis and increased synovial membrane.With aggravation ofsymptoms,newborn blood vessels were increasing. Synovial tissues a fewnewborn blood vessels and infiltrating inflammatory cells as well asslightly increased synovial membrane was showed in BZXD group.Onfollowing 35 days, the newborn blood vessels decreased, with relievinginflammation and thinner synovial membrane. Pathologic injures couldbe ameliorated by BZXD.2. Annexin 1 and aldolase A expression of synovium at differentstages from the normal, model and BZXD group:(1) Annexin 1 expression of synovium at different stages from thenormal, model and BZXD group: The expression of annexin 1 insynovium in model group was notably lower than that in normal group atthe different times(P＜0.05, P＜0.05, P＜0.01).Annexin 1 expression inmodel group was down-regulated step by step (P＜0.05, P＜0.05, P＜0.01),but that of the BZXD group decreased and were significantly higher thanthat in model group(P＜0.01),and was close to that of normal group on 45days (P＞0.05).(2) Aldolase A expression of synovium at different stages from thenormal, model and BZXD group: The expression of aldolase A in modelgroup was notably higher than that in normal group at the different times(P＜0.05, P＜0.01, P＜0.01). Aldolase A expression in model group gradely increased (P＜0.05, P＜0.01, P＜0.01), but that of the BZXDgroup decreased and were significantly lower than that in model group(P＜0.01),and was close to that of normal group on 45 days(P＞0.05).(3) Annexinl is negatively correlated with aldolase A in model andBZXD group.Conclusion:1. BⅡC mixing CFA hyodermic imitates CIA model successfully,that could represent RA for research.. BZXD can relieve the pathologicinjures of CIA rats.2. During the process of CIA, with arthritis more severe, annexin 1edge down and aldolase A edge up. Annexinl is negatively correlatedwith aldolase A. It was indicated annexin 1 and aldolase A are related toCIA synovitis and RA.3. BZXD can edge up annexin 1 expression and edge down aldolaseA expression in synovial tissue of CIA rat,which may be the treatmentmechanism of RA.