【摘要】： 目的:本課題對痛風(fēng)性關(guān)節(jié)炎大鼠模型的制備及關(guān)節(jié)腫脹度檢測方法進行了比較,對未知療效的三種中藥復(fù)方提取物做篩選,對篩選出的最佳中藥復(fù)方提取物JFX-H-50-30作了量效關(guān)系研究,還對其在抑制關(guān)節(jié)炎炎癥反應(yīng)和對高尿酸癥的影響作了研究,為進一步開發(fā)新藥奠定基礎(chǔ)。 方法:(1)采用5%和1.3%兩種不同濃度尿酸鈉注射入大鼠右后足踝的脛跗關(guān)節(jié)腔內(nèi)制備關(guān)節(jié)炎模型并進行對比。(2)采用5%濃度尿酸鈉制備痛風(fēng)性關(guān)節(jié)炎大鼠模型,選用聚乙烯吡咯烷酮(PVP)為溶劑對照、吲哚美辛和已知有抗炎作用的中藥復(fù)方這兩種測試藥物,對周長、直徑和重量這幾種檢測法的優(yōu)缺點進行比較,確定最佳檢測方法。(3)以PVP為溶劑對照、吲哚美辛為陽性藥物,對三種中藥復(fù)方提取物JFX-H-100-60、JFX-H-50-30、JFX-H-50-60進行藥效學(xué)實驗,確定最佳中藥提取物。(4)以PVP作為溶劑對照,吲哚美辛作為陽性藥物,對中藥復(fù)方提取物JFX-H-50-30的六種給藥劑量0.03 g/kg(每kg體重給藥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作藥效學(xué)研究,并作量效分析。(5)將家兔分為三組,分別灌胃給以PVP(溶劑對照)、吲哚美辛(陽性對照),中藥復(fù)方提取物JFX-H-50-30,采用尿酸鈉注入家兔膝關(guān)節(jié)腔造模,收集關(guān)節(jié)腔積液于顯微鏡下進行白細胞計數(shù)。(6)將小鼠分成6組,即正常組、高尿酸模型組(PVP溶劑組)、JFX-H-50-30低、中、高劑量組和別嘌呤醇組,除正常組腹腔注射生理鹽水外,其他組均腹腔注射900 mg/kg的氧嗪酸鉀,小鼠眼眶后靜脈叢采血,用全自動生化儀測定血清中尿酸值并進行比較。 結(jié)果:(1)采用濃度為1.3%的尿酸鈉溶液50μl注射,大鼠足踝腫脹度不夠,造模3 h后腫脹自然消退,不穩(wěn)定,而采用5%尿酸鈉50μl注射,足踝腫脹度在造模后1-6 h一直維持在較高水平。(2)采用直徑法、周長法和重量法對溶劑對照組、吲哚美辛組、中藥復(fù)方組造模后關(guān)節(jié)炎嚴重程度進行測定和比較可知,幾種檢測方法中,從方便性、誤差大小、敏感性、時間過程等各項指標(biāo)綜合考慮,周長法為最佳。(3)對三種中藥復(fù)方提取物的進行初步篩選發(fā)現(xiàn),與空白對照組相比,JFX-H-50-30和JFX-H-50-60有非常顯著抗炎作用(P0.01),JFX-H-100-60無抗炎作用。進行組間比較發(fā)現(xiàn),JFX-H-50-30的抗炎作用優(yōu)于JFX-H-50-60(P0.05)。(4)中藥提取物JFX-H-50-30的1.0 g/kg劑量、0.75 g/kg劑量、0.50 g/kg劑量均有非常顯著抗炎作用(P 0.01),0.30 g/kg劑量和0.10 g/kg劑量組有顯著抗炎作用(P 0.05)。0.03 g/kg劑量沒有抗炎作用。中藥復(fù)方JFX-H-50-30在0.10 g/kg～1.0 g/kg劑量范圍有抗炎作用。(5)家兔關(guān)節(jié)腔積液于顯微鏡下進行白細胞計數(shù)結(jié)果表明,與PVP溶劑對照(模型組)相比,中藥復(fù)方JFX-H-50-30組的關(guān)節(jié)液中白細胞數(shù)量的影響顯著減少(P0.05),說明JFX-H-50-30有抗炎作用。(6)與模型組比較,中藥復(fù)方低、中、高劑量組均非常顯著地降低高尿酸血癥小鼠血清尿酸水平(P0.01,P0.01,P0.01)。 結(jié)論:(1)尿酸鈉的不同濃度會影響造模的程度,實驗顯示選用濃度為5%的尿酸鈉注射較為合理。同時,注射體積也會影響足腫脹度,從大鼠足踝脛跗關(guān)節(jié)腔的容積考慮,50μl的注射量較為合理。(2)檢測足腫脹度的多種方法中,周長法有誤差小、敏感性適中、能反映整體腫脹度和可全程檢測的優(yōu)點,是相對較好的檢測方法。(3)三種中藥復(fù)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的抗炎作用優(yōu)于JFX-H-50-60。JFX-H-100-60無抗炎作用。(4)JFX-H-50-30在0.10 g/kg～1.0 g/kg劑量范圍有抗炎作用并存在量效關(guān)系,0.50 g/kg以上劑量的抗炎作用優(yōu)于0.30 g/kg以下劑量。(5)JFX-H-50-30能使急性痛風(fēng)性關(guān)節(jié)炎家兔炎癥關(guān)節(jié)腔積液中白細胞數(shù)量的影響顯著減少(6)JFX-H-50-30能顯著地降低高尿酸血癥小鼠血清尿酸水平,并存在一定的量效關(guān)系,JFX-H-50-30劑量越大,降尿酸作用越強。(7)JFX-H-50-30對尿酸鈉引起的痛風(fēng)性關(guān)節(jié)炎有抗炎作用,同時也有抗高尿酸血癥的作用,是一個特異性的防治痛風(fēng)藥物,有開發(fā)價值,值得進行深入研究。
[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.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R-332;R285

【引證文獻】

相關(guān)博士學(xué)位論文 前1條

1 王占奎;清瀉濁毒法對實驗性高尿酸血癥和急性痛風(fēng)性關(guān)節(jié)炎的影響及機理研究[D];山東中醫(yī)藥大學(xué);2010年

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本文編號：2236009