蕲蛇醇提液治疗大鼠骨关节炎的药效及作用机制  

Effect of alcohol extract from Agkistrodon acutus on rat pain model of osteoarthritis and its mechanism

作　　者：程佩玲 严波 杜文喜[3] 鲍荣华 陈俊杰[3] CHENG Peiling;YAN Bo;DU Wenxi;BAO Ronghua;CHEN Junjie(Department of Pharmacy,Lishui Hospital of Traditional Chinese Medicine,Lishui 323020,China;不详)

机构地区：[1]丽水市中医院药剂科,323020 [2]浙江中医药大学药学院 [3]浙江中医药大学附属第一医院骨伤科 [4]富阳中医骨伤医院骨伤科

出　　处：《浙江医学》2024年第9期920-925,I0005,共7页Zhejiang Medical Journal

基　　金：浙江省自然科学基金资助项目(LY20H270014)。

摘　　要：目的探究蕲蛇醇提液治疗大鼠骨关节炎(OA)的药效及可能的作用机制。方法30只SD大鼠随机分为正常组、模型组、蕲蛇醇提组,每组10只,除正常组外均采用碘乙酸(MIA)关节腔注射法建立OA疼痛模型。造模7 d后,蕲蛇醇提组大鼠膝关节腔注射0.2 mL蕲蛇醇提液(0.6 g/kg),正常组和模型组大鼠注射等体积0.9%氯化钠注射液,1周1次,连续4周。于给药前和给药4周后测量大鼠机械痛和热痛阈值,通过疼痛行为学评估、组织病理学观察、免疫组化分析、Tunel原位末端凋亡染色和Western blot,评价蕲蛇醇提液对OA大鼠的体内疗效,探索其潜在的作用机制。结果相比正常组,模型组大鼠关节组织表现出软骨退变、软骨细胞数量减少、胶原蛋白大量破坏、基质紊乱的病理状态。相比模型组,蕲蛇醇提液组大鼠病理状态改善,机械痛和热痛的痛阈值显著回升(P<0.01),国际骨关节炎研究协会(OARSI)病理评分显著降低(P<0.01)。免疫组化结果显示,蕲蛇醇提组大鼠关节中Ⅱ型胶原(Col2)阳性率升高,基质金属蛋白酶13(MMP-13)阳性率降低。Tunel检测结果表明,蕲蛇醇提组细胞凋亡率降低(P<0.01)。Western blot结果显示,蕲蛇醇提组MMP-13和聚腺苷二磷酸核糖聚合酶(PARP)及磷酸化p65蛋白相对表达量显著下降(P<0.05)。结论蕲蛇醇提液通过调控PARP/NF-κB通路抑制细胞凋亡,促进软骨代谢平衡,缓解关节疼痛,修复软骨损伤。Objective To explore the efficacy and possible mechanism of Agkistrodon acutus alcohol extract on osteoarthritis(OA).Methods Thirty SD rats were randomly divided into control group,model group and Acutokus acutus alcohol extract group(extract group),with ten rats in each group.The OA pain model was established by the joint injection of iodoacetic acid(MIA).Seven days after modeling,the rats in extract group were injected with 0.2 mL Agkistrodon acutus alcohol extract(0.6 g/kg),and the rats in control and model groups were injected with the same amount of saline,once a week for continuous 4 weeks.Mechanical and thermal pain thresholds were measured before and 4 weeks after administration.Pain behavior evaluation,histopathological observation,immunohistochemical analysis,terminal oxynucleotidyl transferase mediated dUTP biotin nick end labeling(Tunel)staining and Western blot were used to evaluate the in vivo efficacy of Agkistrodon acutus alcohol extract on OA rats and explore its potential mechanism of action.Results The model of OA pain rats was established successfully.Compared with the control group,the joint tissues of model group showed the pathological state of cartilage degeneration,reduced number of chondrocytes,massive destruction of collagen and matrix disorder.Compared with the model group,the pathological state of the rats in extract group was improved and the pain thresholds of mechanical pain and thermal pain increased significantly(P<0.01),while the Osteoarthritis Research Society International(OARSI)scores were significantly lower than those of the model group(P<0.01).Immunohistochemical results showed that in extract group,the positive expression rate of collagen typeⅡ(Col2)in rat joints increased while that of matrix metalloproteinase 13(MMP-13)decreased;Tunel test results showed that cell apoptosis rate was significantly reduced(P<0.01).Based on Western blot,the protein expression levels of MMP-13,poly ADP-ribose polymerase(PARP)and phosphorylated p65(p-p65)in extract group were significantly

关 键 词：蕲蛇醇提液 骨性关节炎 作用机制 

分 类 号：R285.5[医药卫生—中药学]