痛风性关节炎大鼠模型的制备及防治痛风中药复方的研究
发布时间：2018-09-11 07:24
【摘要】： 目的:本课题对痛风性关节炎大鼠模型的制备及关节肿胀度检测方法进行了比较,对未知疗效的三种中药复方提取物做筛选,对筛选出的最佳中药复方提取物JFX-H-50-30作了量效关系研究,还对其在抑制关节炎炎症反应和对高尿酸症的影响作了研究,为进一步开发新药奠定基础。 方法:(1)采用5%和1.3%两种不同浓度尿酸钠注射入大鼠右后足踝的胫跗关节腔内制备关节炎模型并进行对比。(2)采用5%浓度尿酸钠制备痛风性关节炎大鼠模型,选用聚乙烯吡咯烷酮(PVP)为溶剂对照、吲哚美辛和已知有抗炎作用的中药复方这两种测试药物,对周长、直径和重量这几种检测法的优缺点进行比较,确定最佳检测方法。(3)以PVP为溶剂对照、吲哚美辛为阳性药物,对三种中药复方提取物JFX-H-100-60、JFX-H-50-30、JFX-H-50-60进行药效学实验,确定最佳中药提取物。(4)以PVP作为溶剂对照,吲哚美辛作为阳性药物,对中药复方提取物JFX-H-50-30的六种给药剂量0.03 g/kg(每kg体重给药0.03g,下同)、0.10 g/kg、0.30 g/kg、0.50 g/kg、0.75 g/kg和1.0 g/kg作药效学研究,并作量效分析。(5)将家兔分为三组,分别灌胃给以PVP(溶剂对照)、吲哚美辛(阳性对照),中药复方提取物JFX-H-50-30,采用尿酸钠注入家兔膝关节腔造模,收集关节腔积液于显微镜下进行白细胞计数。(6)将小鼠分成6组,即正常组、高尿酸模型组(PVP溶剂组)、JFX-H-50-30低、中、高剂量组和别嘌呤醇组,除正常组腹腔注射生理盐水外,其他组均腹腔注射900 mg/kg的氧嗪酸钾,小鼠眼眶后静脉丛采血,用全自动生化仪测定血清中尿酸值并进行比较。 结果:(1)采用浓度为1.3%的尿酸钠溶液50μl注射,大鼠足踝肿胀度不够,造模3 h后肿胀自然消退,不稳定,而采用5%尿酸钠50μl注射,足踝肿胀度在造模后1-6 h一直维持在较高水平。(2)采用直径法、周长法和重量法对溶剂对照组、吲哚美辛组、中药复方组造模后关节炎严重程度进行测定和比较可知,几种检测方法中,从方便性、误差大小、敏感性、时间过程等各项指标综合考虑,周长法为最佳。(3)对三种中药复方提取物的进行初步筛选发现,与空白对照组相比,JFX-H-50-30和JFX-H-50-60有非常显著抗炎作用(P0.01),JFX-H-100-60无抗炎作用。进行组间比较发现,JFX-H-50-30的抗炎作用优于JFX-H-50-60(P0.05)。(4)中药提取物JFX-H-50-30的1.0 g/kg剂量、0.75 g/kg剂量、0.50 g/kg剂量均有非常显著抗炎作用(P 0.01),0.30 g/kg剂量和0.10 g/kg剂量组有显著抗炎作用(P 0.05)。0.03 g/kg剂量没有抗炎作用。中药复方JFX-H-50-30在0.10 g/kg～1.0 g/kg剂量范围有抗炎作用。(5)家兔关节腔积液于显微镜下进行白细胞计数结果表明,与PVP溶剂对照(模型组)相比,中药复方JFX-H-50-30组的关节液中白细胞数量的影响显著减少(P0.05),说明JFX-H-50-30有抗炎作用。(6)与模型组比较,中药复方低、中、高剂量组均非常显著地降低高尿酸血症小鼠血清尿酸水平(P0.01,P0.01,P0.01)。 结论:(1)尿酸钠的不同浓度会影响造模的程度,实验显示选用浓度为5%的尿酸钠注射较为合理。同时,注射体积也会影响足肿胀度,从大鼠足踝胫跗关节腔的容积考虑,50μl的注射量较为合理。(2)检测足肿胀度的多种方法中,周长法有误差小、敏感性适中、能反映整体肿胀度和可全程检测的优点,是相对较好的检测方法。(3)三种中药复方提取物JFX-H-100-60、JFX-H-50-30、JFX-H-50-60中JFX-H-50-30和JFX-H-50-60有抗炎作用,且JFX-H-50-30的抗炎作用优于JFX-H-50-60。JFX-H-100-60无抗炎作用。(4)JFX-H-50-30在0.10 g/kg～1.0 g/kg剂量范围有抗炎作用并存在量效关系,0.50 g/kg以上剂量的抗炎作用优于0.30 g/kg以下剂量。(5)JFX-H-50-30能使急性痛风性关节炎家兔炎症关节腔积液中白细胞数量的影响显著减少(6)JFX-H-50-30能显著地降低高尿酸血症小鼠血清尿酸水平,并存在一定的量效关系,JFX-H-50-30剂量越大,降尿酸作用越强。(7)JFX-H-50-30对尿酸钠引起的痛风性关节炎有抗炎作用,同时也有抗高尿酸血症的作用,是一个特异性的防治痛风药物,有开发价值,值得进行深入研究。
[Abstract]:OBJECTIVE: To compare the preparation of gouty arthritis rat model and the methods of detecting joint swelling degree, to screen three kinds of Chinese herbal compound extracts with unknown curative effect, and to study the dose-effect relationship between JFX-H-50-30, the best Chinese herbal compound extract, and to inhibit the inflammatory reaction of arthritis and hyperuricemia. The research has laid a foundation for further development of new drugs.
METHODS: (1) Two different concentrations of sodium urate were injected into the tibial tarsal joint cavity of the right hind ankle of rats to establish arthritis models and compare them. (2) Gouty arthritis rat models were established by using 5% concentration of sodium urate, polyvinylpyrrolidone (PVP) as solvent control, indomethacin and the known anti-inflammatory Chinese herbal compound. The advantages and disadvantages of these two methods were compared to determine the best detection method. (3) Using PVP as the solvent control, indomethacin as the positive drug, the pharmacodynamic experiments were carried out on the extracts of three traditional Chinese medicine compounds JFX-H-100-60, JFX-H-50-30, JFX-H-50-60, to determine the best extract of traditional Chinese medicine. (4) Using PVP as the solvent control, indomethacin as the positive drug. Indomethacin was used as a positive drug. The six doses of JFX-H-50-30 were 0.03 g/kg, 0.10 g/kg, 0.30 g/kg, 0.50 g/kg, 0.75 g/kg and 1.0 g/kg, respectively. The rabbits were divided into three groups and given PVP (solvent control), indomethacin and indomethacin respectively. (6) The mice were divided into 6 groups: normal group, hyperuricemia model group (PVP solvent group), JFX-H-50-30 low, middle, high dose group and allopurinol group, except normal group. In addition to normal saline, 900 mg/kg potassium oxalate was intraperitoneally injected into other groups. Blood samples were collected from the posterior orbital venous plexus of mice. The uric acid in serum was determined by automatic biochemical analyzer and compared.
Results: (1) The swelling of ankle and foot in rats was not enough when injected with sodium uric acid solution of 1.3% at 50 ml. The swelling subsided naturally and was unstable after 3 hours of modeling. The swelling of ankle and foot was maintained at a higher level 1-6 hours after modeling. 2) The solvent control group, indomethacin group and indomethacin group were treated with diameter method, perimeter method and weight method. The severity of arthritis in the compound prescription group was measured and compared. Several test methods, including convenience, error size, sensitivity, time process and other indicators, were considered comprehensively, and the perimeter method was the best. (3) The preliminary screening of extracts from three Chinese herbal prescriptions showed that compared with the blank control group, JFX-H-50-30 and JFX-H-50-60. The anti-inflammatory effect of JFX-H-50-30 was better than that of JFX-H-50-60 (P The dose of JFX-H-50-30 was in the range of 0.10 g/kg to 1.0 g/kg. (5) Leukocyte count in rabbit articular effusion under microscope showed that the number of leucocytes in joint fluid of JFX-H-50-30 group was higher than that of PVP group. The effect of JFX-H-50-30 was significantly decreased (P 0.05), indicating that JFX-H-50-30 had anti-inflammatory effect. (6) Compared with the model group, the traditional Chinese medicine compound was lower, the middle and high dose groups were significantly lower than the hyperuricemia mice serum uric acid level (P 0.01, P 0.01, P 0.01).
Conclusion: (1) Different concentrations of sodium urate can affect the degree of modeling. The experiment shows that 5% sodium urate injection is more reasonable. At the same time, the injection volume also affects the degree of foot swelling. Considering the volume of the rat's foot, ankle, tibiotarsal joint cavity, the injection volume of 50 ml is more reasonable. (2) There are errors in the perimeter method in many methods of measuring the degree of foot swelling. JFX-H-50-30, JFX-H-50-30 and JFX-H-50-60 in three Chinese herbal compound extracts, JFX-H-50-30, JFX-H-50-60, JFX-H-50-60 have anti-inflammatory effects, and JFX-H-50-30 has better anti-inflammatory effects than JFX-H-50-60.JFX-H-50-H-100-60. The anti-inflammatory effect of JFX-H-50-30 in the range of 0.10 g/kg to 1.0 g/kg was better than that of 0.30 g/kg. (5) JFX-H-50-30 could significantly reduce the number of leukocytes in inflammatory joint effusion of rabbits with acute gouty arthritis. (6) JFX-H-50-30 could significantly reduce hyperuricemia. The higher the dose of JFX-H-50-30, the stronger the effect of reducing uric acid. (7) JFX-H-50-30 has anti-inflammatory effect on gouty arthritis caused by sodium urate, and also has Anti-hyperuricemia effect. It is a specific drug for gout prevention and treatment. It is worth further study.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R-332;R285
本文编号：2236009
[Abstract]:OBJECTIVE: To compare the preparation of gouty arthritis rat model and the methods of detecting joint swelling degree, to screen three kinds of Chinese herbal compound extracts with unknown curative effect, and to study the dose-effect relationship between JFX-H-50-30, the best Chinese herbal compound extract, and to inhibit the inflammatory reaction of arthritis and hyperuricemia. The research has laid a foundation for further development of new drugs.
METHODS: (1) Two different concentrations of sodium urate were injected into the tibial tarsal joint cavity of the right hind ankle of rats to establish arthritis models and compare them. (2) Gouty arthritis rat models were established by using 5% concentration of sodium urate, polyvinylpyrrolidone (PVP) as solvent control, indomethacin and the known anti-inflammatory Chinese herbal compound. The advantages and disadvantages of these two methods were compared to determine the best detection method. (3) Using PVP as the solvent control, indomethacin as the positive drug, the pharmacodynamic experiments were carried out on the extracts of three traditional Chinese medicine compounds JFX-H-100-60, JFX-H-50-30, JFX-H-50-60, to determine the best extract of traditional Chinese medicine. (4) Using PVP as the solvent control, indomethacin as the positive drug. Indomethacin was used as a positive drug. The six doses of JFX-H-50-30 were 0.03 g/kg, 0.10 g/kg, 0.30 g/kg, 0.50 g/kg, 0.75 g/kg and 1.0 g/kg, respectively. The rabbits were divided into three groups and given PVP (solvent control), indomethacin and indomethacin respectively. (6) The mice were divided into 6 groups: normal group, hyperuricemia model group (PVP solvent group), JFX-H-50-30 low, middle, high dose group and allopurinol group, except normal group. In addition to normal saline, 900 mg/kg potassium oxalate was intraperitoneally injected into other groups. Blood samples were collected from the posterior orbital venous plexus of mice. The uric acid in serum was determined by automatic biochemical analyzer and compared.
Results: (1) The swelling of ankle and foot in rats was not enough when injected with sodium uric acid solution of 1.3% at 50 ml. The swelling subsided naturally and was unstable after 3 hours of modeling. The swelling of ankle and foot was maintained at a higher level 1-6 hours after modeling. 2) The solvent control group, indomethacin group and indomethacin group were treated with diameter method, perimeter method and weight method. The severity of arthritis in the compound prescription group was measured and compared. Several test methods, including convenience, error size, sensitivity, time process and other indicators, were considered comprehensively, and the perimeter method was the best. (3) The preliminary screening of extracts from three Chinese herbal prescriptions showed that compared with the blank control group, JFX-H-50-30 and JFX-H-50-60. The anti-inflammatory effect of JFX-H-50-30 was better than that of JFX-H-50-60 (P The dose of JFX-H-50-30 was in the range of 0.10 g/kg to 1.0 g/kg. (5) Leukocyte count in rabbit articular effusion under microscope showed that the number of leucocytes in joint fluid of JFX-H-50-30 group was higher than that of PVP group. The effect of JFX-H-50-30 was significantly decreased (P 0.05), indicating that JFX-H-50-30 had anti-inflammatory effect. (6) Compared with the model group, the traditional Chinese medicine compound was lower, the middle and high dose groups were significantly lower than the hyperuricemia mice serum uric acid level (P 0.01, P 0.01, P 0.01).
Conclusion: (1) Different concentrations of sodium urate can affect the degree of modeling. The experiment shows that 5% sodium urate injection is more reasonable. At the same time, the injection volume also affects the degree of foot swelling. Considering the volume of the rat's foot, ankle, tibiotarsal joint cavity, the injection volume of 50 ml is more reasonable. (2) There are errors in the perimeter method in many methods of measuring the degree of foot swelling. JFX-H-50-30, JFX-H-50-30 and JFX-H-50-60 in three Chinese herbal compound extracts, JFX-H-50-30, JFX-H-50-60, JFX-H-50-60 have anti-inflammatory effects, and JFX-H-50-30 has better anti-inflammatory effects than JFX-H-50-60.JFX-H-50-H-100-60. The anti-inflammatory effect of JFX-H-50-30 in the range of 0.10 g/kg to 1.0 g/kg was better than that of 0.30 g/kg. (5) JFX-H-50-30 could significantly reduce the number of leukocytes in inflammatory joint effusion of rabbits with acute gouty arthritis. (6) JFX-H-50-30 could significantly reduce hyperuricemia. The higher the dose of JFX-H-50-30, the stronger the effect of reducing uric acid. (7) JFX-H-50-30 has anti-inflammatory effect on gouty arthritis caused by sodium urate, and also has Anti-hyperuricemia effect. It is a specific drug for gout prevention and treatment. It is worth further study.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R-332;R285
【引证文献】
相关博士学位论文 前1条
1 王占奎;清泻浊毒法对实验性高尿酸血症和急性痛风性关节炎的影响及机理研究[D];山东中医药大学;2010年
，本文编号：2236009
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2236009.html
最近更新
教材专著
