柔性纳米脂质体载体增强双氯芬酸钠的经皮渗透效果:与普通外用制剂比较的随机对照动物实验

Enhancement in the percutaneous permeation effects of diclofenac sodium by nanoliposome carrier:A comparative randomized study with common external preparation

目的:通过与普通外用制剂对比,观察双氯芬酸钠柔性纳米脂质体对类风湿性关节炎大鼠的镇痛、抗炎效果,评估柔性纳米脂质体这种新型皮肤给药载体的促进药物局部吸收作用。方法:实验于2005-03/11在中南大学湘雅医院中西结合研究所完成。选择SD大鼠80只,按随机数字表法分成4组:正常对照组、扶他林组、5%双氯芬酸钠柔性纳米脂质体组、10%双氯芬酸钠柔性纳米脂质体组,每组20只。每组10只用于镇痛实验,10只用于抗炎实验。①镇痛实验:热板实验:每组大鼠双后足分别涂抹生理盐水、扶他林乳胶剂、5%双氯芬酸钠柔性纳米脂质体和10%双氯芬酸钠柔性纳米脂质体,每足0.5g,每隔6min涂药1次,连续5次。末次给药10min后将足洗净,在5,10,15,25min分别测定大鼠舔足次数。扭体实验:每组大鼠腹部分别涂抹生理盐水、扶他林乳胶剂、5%双氯芬酸钠柔性纳米脂质体和10%双氯芬酸钠柔性纳米脂质体,给药20min后,各组大鼠分别腹腔注射0.6%的醋酸溶液,0.01mL/g,分别在给药后1,2,4,6h观察并记录20min内大鼠的扭体次数,计算其扭体反应抑制率,扭体反应抑制率=[(正常对照组扭体次数-用药组扭体次数)/正常对照组扭体次数]×100%。甲醛舔足实验:每组大鼠右后足分别涂抹生理盐水、扶他林乳胶剂、5%双氯芬酸钠柔性纳米脂质体和10%双氯芬酸钠柔性纳米脂质体,每10min涂药1次。末次给药10min后分别于右后足皮下注射2.5%甲醛溶液30μL,分别记录每只大鼠10min内舔足的次数。②抗炎实验:扶他林组、5%双氯芬酸钠柔性纳米脂质体组、10%双氯芬酸钠柔性纳米脂质体组采用皮下注射牛Ⅱ型胶原和完全福氏佐剂制备类风湿性关节炎模型。正常对照组不予任何处理。每组大鼠右后足分别涂抹生理盐水、扶他林乳胶剂,5%双氯芬酸钠柔性纳米脂质体和10%双氯芬酸钠柔性纳米脂质体,约0.5～1.0g/次(视关节大小而定,即每次给药量按浓度折算为临床常用量100～150mg/d),3次/d。测量致炎后7,l4,21,28,35d右后足踝关节周长。结果:纳入大鼠80只,均进入结果分析。扶他林组、5%双氯芬酸钠柔性纳米脂质体组和10%双氯芬酸钠柔性纳米脂质体组镇痛、抗炎效果明显高于正常对照组,差异有显著性意义(P<0.01);5%双氯芬酸钠柔性纳米脂质体组和10%双氯芬酸钠柔性纳米脂质体组镇痛、抗炎效果明显高于扶他林组,差异有显著性意义(P<0.01);5%双氯芬酸钠柔性纳米脂质体组与10%双氯芬酸钠柔性纳米脂质体组镇痛、抗炎效果差异无显著性意义(P>0.05)。结论:外用不同剂量双氯芬酸钠柔性纳米脂质体对类风湿性关节炎大鼠较普通外用制剂在镇痛、抗炎方面有更明显的疗效。

AIM： To investigate the analgesic and anti-inflammatory effects of the nanoliposomes loaded with diclofenac sodium to the rats with rheumatoid arthritis （RA）, and evaluate the local drug absorption improved by nanoliposomes as a new skin administration carder. METHODS： The experiment was completed in the Institute of Combined Traditional Chinese and Western Medicine, Xiangya Hospital of Central South University from March through November in 2005. Totally 80 SD rats were randomly divided into 4 groups： normal control group, voltaren group, and other two treatment groups containing 5% and 10% of nanoliposomes load with diclofenac sodium, respectively. Each group consisted of 20 rats, including 10 rats for analgesic test and 10 for anti-inflammatory test. （1）Analgesic tests： Hot-plate test was conducted by painting corresponding drugs on double behind feet, 0.5 g each foot in 6-minute interval for 5 times continuously. At 5, 10, 15, 25 minutes after the last treatment, the times of licking foot were examined. Writhing test was conducted by painting corresponding drugs on abdominal skin, 20 minutes later, intraperitoneal injection of 0.6% acetic acid solution was given 0.01 mL/g. At 1, 2, 4, 6 hours after the treatment, the times of writhing was recorded within 20 minutes and the inhibition ratio of writhing was calculated according to [（writhing times of normal control group- writhing times of treatment group）/writhing times of normal control group]×100%. FormaSn-induced licking test was conducted by painting corresponding drugs on right behind feet, once 10 minutes, which was followed by subcutaneous injection of 2.5% formalin, 30 μL. Then the times of licking foot was recorded within 10 minutes.（2） Anti-inflammatory test： RA model was established by collagen Ⅱ and complete Freund＇s adjuvant in three treatment groups, while no management was given for the normal control group. Rats in each group were painted with corresponding drugs in the right behind feet, about 0.5-1.0 g （according to the size of the articles, clinical dose was defined as 100-150 mg/d）, three times daily. The perimeter of the ankle joint of the model rats＇ right behind feet was measured in the 7^th, 14^th, 21^st, 28^th, 35^th days. RESULTS： All of 80 rats were enrolled in the final analysis. Compared with normal group, other 3 treatment groups investigated had relatively good analgesic and anti-inflammatory effects. Among them, better analgesic and anti-inflammatory effects were found in the groups containing the drug-loaded nanoliposomes, in particular in the group containing 10% of the drug-loaded nanoliposomes. CONCLUSION： The .use of the nanoliposomes as the carrier for diclofenac sodium improves the analgesic and anti-inflammatory effects.