蕲蛇水提液对大鼠骨性关节炎疼痛模型的作用研究

Study on the Effect of Agkistrodon Aqueous Extract on Pain Model of Osteoarthritis in Rats

关键词:  蕲蛇  骨性关节炎  碘乙酸  水提液

Key words:  Agkistrodon  osteoarthritis  iodoacetic acid  aqueous extract

摘要:

目的 研究蕲蛇水提液对大鼠骨性关节炎（osteoarthritis，OA）的药效作用。方法 将SD大鼠随机分为正常组、模型组、蕲蛇组，每组10只，除正常组外均采用碘乙酸关节腔注射法（50 μL）建立OA疼痛模型。造模48 h后，蕲蛇组大鼠关节腔注射蕲蛇水提液（100 μg·mL^-1），模型组大鼠注射等量生理盐水，1周1次，连续4周。每周测量大鼠足耐压痛、热痛阈的数值变化，观察大鼠膝关节的疼痛改善情况；4周后取关节进行病理切片SO染色，观察膝关节的组织病理改变。培养大鼠原代软骨细胞，以CCK-8法评价蕲蛇对软骨细胞的增殖作用。结果 OA大鼠造模成功。与模型组相比，蕲蛇组压痛和热痛的痛阈值显著回升（P<0.01）；软骨基本恢复正常，软骨细胞和软骨基质接近正常水平，仅有少量肥大软骨细胞；Mankin评分显著降低（P<0.01）；软骨细胞活力显著升高（P<0.01），各项指标基本到达正常水平。结论 蕲蛇水提液可显著改善OA大鼠关节疼痛，并修复软骨损伤，其机制与促进软骨细胞增殖有关。

Abstract:

OBJECTIVE To study the pharmacological effect of Agkistrodon aqueous extract on osteoarthritis(OA) in rats. METHODS SD rats were randomly divided into 3 groups:normal group, model group and Agkistrodon group, 10 rats in each group. Except the normal group, the OA pain model was established by using the iodoacetic acid to inject the joint cavity(50 μL). After 48 h, the Agkistrodon group were injected with Agkistrodon aqueous extract(100 μg·mL^-1) in articular, and the model group were injected with the same amount of saline, once a week, for four consecutive weeks. The numerical changes of the paw withdrawal themal latency of the rats were measured every week, and observed the pain of the knee joint. After 4 weeks, the joint was taken for pathological section with staining so that the pathological changes of the knee joint were observed. The rat chondrocytes were cultured and the effect of Agkistrodon aqueous extract on the proliferation of cartilage cells was evaluated by CCK-8 method. RESULTS The OA pain model was established successfully. Compared with model group, the threshold of pressing pain and heat pain in Agkistrodon group were significantly increased(P<0.01); cartilage basically returned to normal, chondrocytes and cartilage matrix closed to normal levels, only had a small amount of hypertrophic chondrocytes; Mankin score significantly decreased(P<0.01); the chondrocyte viability significantly increased(P<0.01). The above index in Agkistrodon group were closed to normal group. CONCLUSION Agkistrodon acutus aqueous extract can significantly ameliorate joint pain and repair cartilage injury in OA rats. The mechanism is related to the promotion of cartilage cell proliferation and can be used in clinical OA auxiliary intervention.