四妙凉血颗粒5种成分含量测定及对急性痛风性关节炎大鼠的抗炎作用研究  

Determination of the Contents of Five Components in Simiao Liangxue Granules and Its Anti-inflammatory Effects on Acute Gouty Arthritis Rats

作　　者：唐春丽[1] 农必华[1] 冯茵怡 林忆龙 魏江存 罗远[1] TANG Chunli;NONG Bihua;FENG Yinyi;LIN Yilong;WEI Jiangcun;LUO Yuan(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Guangxi International Zhuang Medicine Hospital,Nanning 530201,China)

机构地区：[1]广西中医药大学第一附属医院,广西南宁530023 [2]广西中医药大学附属国际壮医医院,广西南宁530201

出　　处：《西部中医药》2025年第2期1-5,共5页Western Journal of Traditional Chinese Medicine

基　　金：国家中医药管理局“十二五”中医药重点学科(民族药学)壮药学资质项目;全国中药特色技术传承人才培训项目(20184828005);广西自然科学基金项目(2020GXNSFAA259059);广西中医药大学高层次人才培育创新团队资助项目(2022A008)。

摘　　要：目的:建立四妙凉血颗粒中5种成分的含量测定方法,探讨四妙凉血颗粒对急性痛风性关节炎(acute gouty arthritis,AGA)大鼠的抗炎作用机制。方法:采用高效液相色谱法测定四妙凉血颗粒中5种成分含量;大鼠足踝关节注射尿酸钠悬液诱导AGA,采用容积法检测足踝关节肿胀程度;采用酶联免疫吸附试验法测定大鼠血清及关节液中白细胞介素1β(interleukin-1β,IL-1β)、IL-6和肿瘤坏死因子α(tumor necrosis factorα,TNF-α)含量;采用免疫组织化学和图像分析方法观测大鼠踝关节软骨组织中TNF-α蛋白表达水平。结果:方法学考察结果显示各项指标符合规定,四妙凉血颗粒样品中落新妇苷、盐酸小檗碱、山柰酚、黄芩素和汉黄芩素的平均回收率为97.14%~102.67%,RSD值均小于3.0%。尿酸钠混悬液注射后,与空白对照组比较,模型组不同时间点大鼠足踝关节肿胀度,关节液中IL-1β、IL-6、TNF-α含量,血清IL-6水平及TNF-α蛋白表达均增高(P<0.05);空白对照组大鼠软骨细胞可见少量TNF-α蛋白表达。与模型组比较,阳性组(秋水仙碱0.65 mg/kg)及四妙凉血颗粒高、中、低剂量(6、3、1.5 g/kg)组大鼠第3~5天足踝关节肿胀度,关节液中IL-1β、IL-6、TNF-α含量及血清IL-6、TNF-α水平均降低(P<0.05);四妙凉血颗粒低、中、高剂量组大鼠TNF-α蛋白积分吸光度降低(P<0.05)。与阳性组比较,四妙凉血颗粒高、中、低剂量组大鼠足踝关节肿胀度,关节液中IL-1β、IL-6、TNF-α含量及血清IL-6、TNF-α表达水平和关节组织TNF-α蛋白积分吸光度均变化不明显,差异无统计学意义(P>0.05)。结论:本实验建立了四妙凉血颗粒含量测定方法,该方法专属性强、重复性好、准确度高,可用于四妙凉血颗粒样品中落新妇苷、盐酸小檗碱、山柰酚、黄芩素和汉黄芩素的含量测定;四妙凉血颗粒治疗AGA的作用机制可能与其降低关节组织炎症因子IL-1β、IL-6和TNF-α水平Objective:To establish the determination methods for the contents of five components in Simiao Liangxue granules and to discuss the anti-inflammatory mechanism of the medicine on rats with acute gouty arthritis(AGA).Methods:HPLC was used to detect the contents of five components in the medicine;AGA rat models were prepared by injecting sodium urate suspension into foot and ankle,the swelling degree of ankle joint was detected by volume method;ELISA method was adopted to measure the contents of IL-1β,IL-6 and TNF-αin the serum and joint fluid;immunohistochemical and image analysis methods were utilized to observe the expressions of TNF-αprotein in cartilage tissue of ankle.Results:The results of methodological examination showed that all the indicators conformed to the regulations,average recovery rates of astilbin,berberine hydrochloride,kaempferol,baicalein and scutellarin ranged between 97.14%and 102.67%in the samples of the medicine,RSD values were smaller than 3.0%.After the injection of sodium urate suspension,compared with blank control group,more significant swelling of ankle joint could be seen,the levels of IL-1β,IL-6 and TNF-αin the joint fluid,the expressions of IL-6 and TNF-αin the serum were elevated in the model group at different time points(P<0.05);small amounts of TNF-αprotein expressions could been found in the blank control group.Compared with the model group,the swelling of ankle joints in the positive group(colchicin,0.65 mg/kg),high,moderate and low doses groups of Simiao Liangxue granules(6,3 and 1.5 g/kg)was alleviated from the third to the fifth day,the contents of IL-1β,IL-6 and TNF-αin joint fluid,the levels of IL-6 and TNF-αin the serum were reduced(P<0.05);TNF-αprotein integral absorbance was lowered in the low,moderate and high doses groups of the medicine(P<0.05).Compared with the positive group,the swelling of ankle joints,the contents of IL-1β,IL-6 and TNF-αin joint fluid,the levels of IL-6 and TNF-αin the serum,as well as TNF-αprotein integral absorbance in the joi

关 键 词：急性痛风性关节炎 四妙凉血颗粒 白细胞介素1Β 白细胞介素6 肿瘤坏死因子α 

分 类 号：R259[医药卫生—中西医结合]