基于信息熵理论比较不同剂量雪上一枝蒿总生物碱对CIA模型大鼠的改善作用及其机制

目的:考察不同剂量雪上一枝蒿总生物碱(简称"雪总碱")对胶原诱发关节炎(CIA)模型大鼠血清炎症因子及关节滑膜组织中FOS蛋白表达的影响,并探讨其抗类风湿关节炎(RA)的可能机制。方法:将雄性SD大鼠随机分为空白组、模型组、阳性组(复方醋酸地塞米松软膏,0.2 g/kg)以及雪总碱低、中、高剂量组(56.26、112.50、225.00 mg/kg,以提取物中雪总碱的质量计),每组10只。除空白组外,其余各组大鼠均于左足皮下注射经不完全费氏佐剂乳化的牛Ⅱ型胶原蛋白以复制CIA模型,并自造模当日起于其左足涂抹相应药物,空白组和模型组大鼠涂抹等容65%乙醇,每日3次,连续28 d。于给药第7、14、21、28天时用游标卡尺测量各组大鼠左后足趾厚度,并计算足肿胀度;于末次给药后采用酶联免疫吸附测定法检测各组大鼠血清白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)含量,采用免疫组化法检测其关节滑膜组织中FOS蛋白的表达水平[以免疫组化评分(HIS)表示];借助熵权法计算综合评分,并以此评价不同给药剂量对CIA模型大鼠上述指标的影响。结果:与空白组比较,模型组大鼠各时间点的足肿胀度、血清炎症因子含量以及HIS值均显著升高(P<0.05)。与模型组比较,各给药组大鼠各时间点足肿胀度,阳性组和雪总碱高剂量组血清IL-1β、IL-6、TNF-α含量以及HIS值,雪总碱中剂量组血清L-6、TNF-α含量以及雪总碱低剂量组血清TNF-α含量均显著降低(P<0.05)。各给药组上述指标的综合评分分别为0.37(阳性组)、0.31(雪总碱高剂量组)、0.23(雪总碱中剂量组)、0.09(雪总碱低剂量组)。结论:雪总碱对CIA模型大鼠具有一定的改善作用,且有随剂量升高而作用增强的趋势。这种作用可能与降低大鼠血清中炎症因子的含量以及抑制关节滑膜组织中FOS蛋白的表达有关。

Comparison of Improvement Effects and Its Mechanism of Different Doses of Total Alkaloids from Aconitum racemulosum on CIA Model Rats Based on Information Entropy Theory

OBJECTIVE:To investigate the effects of different doses of total alkaloids from Aconitum racemulosum(ARTA)on serum inflammation factors and FOS protein expression in synovial tissue of joint in collagen-induced arthritis(CIA) model rats,and to investigate its potential mechanism of anti-rheumatoid arthritis(RA). METHODS:Male SD rats were randomly divided into blank group, model group, positive group(Compound dexamethasone acetate ointment, 0.2 g/kg),ARTA low-dose,medium-dose and high-dose groups(56.26,112.50,225.00 mg/kg,by the weight of ARTA in the extract),with 10 rats in each group. Except for blank group,other groups were given subcutaneous injection of Bovine collagen Ⅱ emulsified with incomplete Freund’s adjuvant into the left foot to establish CIA model;the left foot were smeared with relevant medicine from the day of modeling. Blank group and model group were smeared with constant volume of 65% ethanol,3 times a day,for consecutive 28 days. On the 7 th,14 th,21 st and 28 th day of administration,the thickness of left hind toe was measured with vernier caliper,and the degree of foot swelling was calculated. The serum contents of IL-1β,IL-6 and TNF-α in rats were measured by ELISA after last administration. The expression of FOS protein in synovial tissue was determined by immunohistochemical method [expressed by HIS]. The comprehensive score was conculated by entropy weight method. Effects of each dosage on above indexes of CIA model rats were evaluated with the comprehensive score. RESULTS:Compared with blank group,the degree of foot swelling,serum content of inflammatory factors and HIS value were increased significantly in model group(P<0.05). Compared with model group,the degree of foot swelling in each administration group,serum contents of IL-1β,IL-6 and TNF-α,HIS in positive group and ARTA high-dose group,serum contents of IL-6 and TNF-α in ARTA medium-dose group as well as serum content of TNF-α in ARTA low-dose group were decreased significantly(P<0.05). Comprehensive score of above indicators were 0.37(positive group),0.31(ARTA high-dose group),0.23(ARTA medium-dose group)and 0.09(ARTA low-dose group). CONCLUSIONS:ARTA canimprove CIA model rats,and the effect tends to increase with the increase of dose. Above effect may be associated with reducing serum content of inflammatory factors and inhibiting the expression of FOS protein in synovial tissue.