萆薢牛膝总皂苷防治痛风及其机制研究
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摘要
第一部分 总皂苷的提取及含量测定
目的 从牛膝、萆薢中分别提取总皂苷,测定提取物中总皂苷含量,了解提取物用于药效学试验的可行性。
方法 牛膝切片采用水提、石油醚脱脂、醇提、饱和正丁醇萃取、过氧化铝柱提取总皂苷,采用紫外—可见分光光度法,以齐墩果酸为对照品测定牛膝提取物中总皂苷的含量。萆薢切片乙醇提取、石油醚脱脂、饱和正丁醇萃取、加丙酮沉淀提取总皂苷,采用紫外—可见分光光度法,以薯蓣皂苷元为对照品测定测定萆薢提取物中总皂苷的含量。
结果 齐墩果酸在2.77 μg～13.85 μg范围内线性关系良好,平均加样回收率为97.0%,牛膝提取物中总皂苷含量为80.%。薯蓣皂苷元在5.12 μg～25.6 μg范围内线性关系良好,样品平均加样回收率为97.5%。萆薢提取物中总皂苷含量为53.1%。
结论 上述提取总皂苷方法可行,稳定,总成分含量能满足药效学试验的要求。
第二部分 萆薢总皂苷+牛膝总皂苷的组方合理性研究
目的 从降低血尿酸水平和抗炎作用两方面探讨牛膝总皂苷和萆薢总皂苷的组方合理性及剂量配比。
方法 采用尿酸型小鼠高尿酸血症模型和酵母膏致小鼠高尿酸血症模型以及小鼠耳肿胀模型和微晶型尿酸钠致大鼠急性痛风性关节炎模型模型。按照Codrug软件,得优化组方。
结果 在尿酸型小鼠高尿酸血症模型和酵母膏致小鼠高尿酸血症模型,萆薢总皂苷组、牛膝总皂苷+萆薢总皂苷组和阳性对照组小鼠血清尿酸水平显著降低(P<0.01),牛膝总皂苷480、240、120 mg.kg~(-1)组小鼠血清尿酸水平与模型组比较无显著性差异。萆薢总皂苷组与牛膝总皂苷+萆薢总皂苷组相比小鼠血清尿酸水平也无明显差异。牛膝总皂苷、萆薢总皂苷对小鼠耳肿胀均有显著的抑制作用,牛膝总皂苷+萆薢总皂苷组耳肿胀抑制率均强于单药组。
Part 1 Extracting of total saponin and research on their qualityAim To extract the total saponin of Dioscorea and Achyranthes and research their quality. Methods The total saponin of Achyranthes bidentata (TSA) was extracted by water, alcohol, n-butanol.With oleanolic acid as control, the content of total saponin was determinated by UV. Total saponin of Dioscorea (TSD) was extracted by alcohol, n-butanol. With diosgenin as control, the content of total saponin was determinated by UV.Results The linear relationship of oleanolic acid was better between 2.77μg~13.85μg, average recovery rate was 97% and the content of total saponin of Achyranthes was 80%. The linear relationship of diosgenin was better between 5.12μg~25.6μg, average recovery rate was 97.5% and the content of total saponin was Dioscorea 53.1%.Conclusion The methods was well done to extract total saponin and stable. The ingrements comformed to the rules of animal experiments.Part 2 Study on the rationality of TSD plus TSAAim To study the rationality and the dose of TSD and TSA according to the decrease ofserum uric acid level and anti-inflammatory of TSD and TSA.Methods Mice hyperuricemic models were made by uric acid ip or yeast extract paste ig. Inthese models the effect of TSD, TDA on uric acid level was observed. And mice ear swellingwas induced by dimethylbenzene, rat feet swelling was induced by MSU. In these modelsanti-inflammatory effect of TSD, TSA was measured. According to Codrug software, thegrading-up combination prescription was obtained.Results In the mice hyperuricemic models, the serum uric acid level of TSD group, TSDplus TSA group and positive control group was significantly reduced compared with the
model group(P < 0. 01) .The serum uric acid level of TSA group has no difference from that of the model group. The serum uric acid level of TSD group has no difference from that TSD plus TSA group yet. Both of TSD and TSA could markedly inhibit the mice ear swelling. Moreover the inhibitory of the group of TSD plus TSA was more stronger than the group of TSD or TSA. In the mice ear swelling model, TSA120 mg-kg'1 plus TSD 240mg. kg"1 could reach the maximal inhibitory effect; in the rat feet swelling model TSA 60 mg-kg"1 plus TSD 120 mg. kg"1 could reach the maximal inhibitory effect.Conclusion TSD could reduce the mice serum uric acid level, but TSA could not. Both TSD and TSA had anti-inflammatory effect. TSD plus TSA had more anti-inflammatory effect. In these models the best proportion of TSA and TSD was 1:2.Part 3 Establish animal hyperuricemia model and studies on the effects and mechanisms of TSD A on animal experimental hyperuricemiaAim To establish animal hyperuricemia model and studies on the effects and mechanisms of TSDA on animal experimental hyperuricemia.Methods (l)Mouse was given different dose of uric acid by intraperitoneal injection(ip), the serum uric acid level was detected after 1 h later. (2) Yeast extract paste 7. 5—30 g ? kg"1 was given to mice by ig once daily for 7,14,21,28 consecutive days, and detected the level of uric acid in serum.(3)Mouse and rat hyperuricemic models were made by orally administering yeast extract paste once per day (30g.kg'], 20g.kg"', respectively) for 7 days; another mouse hyperuricemia model was generated by intraperitoneal injection once with uric acid 250 mg.kg"1 or potassium oxonate 300 mg-kg"1. (4)The concentration of uric acid in serum or urine was detected by the phosphotungstic acid method, and the activity of xanthine oxidase (XOD) was assayed by a test kit.Results (1) Uric acid 125, 250, 500, 1000 mg.kg1 ip could significantly increase the level of uric acid in mouse serum. The level of uric acid in mouse serum was attained peak at 10
min and the hyperuricemia could lasted over 4 hours after uric acid 250mg.kg"Igiven. (2)The serum uric acid level in mice were (271. 8±53. 2), (215. 4+31. 5), (195. 9 + 56. 0), (142.1 ±30. 7) umol ? L"1 in 30 g - kg"1 group; and (226. 8±40. 7), (148. 67±30. 4), (176. 9+27.0), (119.3±27. 4) umol . L"1 in 15 g . kg'1 group after administration 1, 2, 3, 4 weeks; In 7. 5 g ? kg'1 group the serum uric acid level was (117. 0+29.0) umol ? L"1 after 1 week. However the serum uric acid of the control group was (108.1 ± 13.9) umol ? L"1. (3)In the mouse hyperuricemia model induced by uric acid, the serum uric acid of the normal group, model group m,TSD plus TSA group (400 ^ 120n 40 mg. kg"1), benzbromarone group (20 mg. kg"1) and allopurinol group (40 mg. kg"1) were 114. 3+21. 4, 229.6+46.9, 160.4 + 32.0, 162.5 + 27.5, 25. 4 ±35. 5, 126.9 ±19.8, 189. 6 ± 31. 6 u mol. L1 respectively.(4) In the mouse hyperuricemia model induced by yeast extract paste, the serum uric acid level were 118.6±29.2, 244.7±47.2, 161.7±34.6, 144.8±31.0, 175.6± 23.9, 81.8±20. 7, 73.0±19.4umol.L~1 respectively. (5) In the mouse hyperuricemia model induced by potassium oxonate, the serum uric acid level were 98. 2+29.9,179. 7± 39.0,131. 7 ± 20.0,140.8 ± 29.2,160. 6 ± 28. 2,88. 7 ± 21.2,117. 3 ± 32.3 u mol. L"1 respectively. (6) In the rat hyperuricemia model induced by yeast extract paste, the serum uric acid level were 152.6+27.9, 282.2±65. 2, 195. 3±66.3, 190. 7±35.8, 243.8+51.1, 178.6 ±40. 7, 121.3 ±57.3 umol.L"1 respectively; the serum XOD activity were 33. 6±3. 2, 52. 4±4. 4, 42. 3±5. 9, 44. 9±5. 1, 44. 6±6. 7, 48. 8±7. 0, 10.8+2. 2 U. I/1 respectively; 24 h total uric acid excretion, were 23. 5 ±4.82, 43.1 +9.18, 41. 4 ±8.53, 35.3±6. 7, 36. 7±8. 06, 43.8±12. 6, 19. 2±4. 9 mmol respectively.Conclusion (1) Uric acid given by ip could form mice hyperuricemia model, the dosage of 250 mg .kg"1 was better. (2) Yeast extract paste (15~30) g ? kg"1 given by ig could form mice hyperuricemia model, and the hyperuricemia could last 1 ~2 weeks.(3) TSDA possesses a potent anti-hyperuricemic effect on hyperuricemic animals, and the mechanism may be relevant in accelerating the excretion and decreasing the production of uric acid. Part 4 Eeffects and mechanisms of TSDA on gouty arthritis of
rabbit and ratAim To observe the preventive and therapeutic effects and mechanisms of TSD on animal gout arthritis.Methods Rat gouty arthritis model and rabbit gouty arthritis model were established by MSU intraarticular injection, the joint swell degree, inflammatory factor and cytokine in the joint fluid or leachate were detected, moreover histopathologic examination was done, rat feet swelling was induced by carrageenan, mice ear swelling was induced by dimethylbenzene, the mice writhing responewas induced by acetic acid, the mice pain induced by warm bath or hot plate, and rat granuloma induced by cotton ball subcutaneously embedding. In these models the swelling degree, pain response latency, granuloma weight and so on were detected respectively.Results hi the rat model, the joint swelling was very obviouse 6 h later and the swelling degree reached the peak at 24h or 48h.The level of NO> TNF-ou PGE2. IL-lp\ IL-6 and IL-8 in the model group was significantly increased and histopathologic examination showed that the synovial cells were seriously injured in the model group, there are many polycyte, histoleucocyte and urete crystal in the joint and soft tissue. TSDA 200, 60mg.kg'1 could significantly inhibit joint swelling of rat and significantly improve the histopathologic changes. TSDA 200mg.kg"1 could improve the gaits and significantly inhibit the increase of NO. TNF- a and PGE2 content as well as the content of IL-6.In the rabbit model, the rabbit joint of the model group appeared red, swelling and higher skin temperature; the fluid increased; WBC in the blood and fluid rose significantly in the model group. The level of NCK TNF-ou PGE2> IL-lp\ IL-6 and IL-8 in the model group was significantly increased and histopathologic examination showed that the synovial cells and soft tissue were seriously injured in the model group. TSDA 100 mg.kg"1 could significantly reduce the fluid in the joint and 100, 30, 10 mg. kg"1 could significantly reduce WBC in the joint fluid. 100, 30 mg.kg"1 inhibited the content of IL-lfi. IL-6 and IL-8.In the model induced by carrageenan, the swellling degree of TSDA 200,60mg.kg" groups was significantly decreased compared with the model group. In the model of cotton
ball granuloma, the weight of granuloma groups were significantly reduced. In the modelsinduced by dimethylbenzen and writhing respone was induced by acetic acid, TSDA400,120,40 mg.kg'1 could significantly reduce mouse ear swelling and frequency of writhingrespone. TSDA 400 mg.kg"! could prolong pain response latency.Conclusion TSDA possessed antigout arthritis effects, its mechanism may be related toinhibit the product of cytokines.Part 5 Effects of TSDA on cytokine secrection and their gene expression of J774 cells and human peripheral blood mononuclear cellsAim To investigate the effects of TSDA on cytokine secrection and their gene expression of J774 cells and human peripheral blood mononuclear cells (PBMC) in vitro. The effect of TSDA on neutrophil chemotaxis was also observed.Methods Isolated total RNA from different groups of J774 cells and PBMC, TNF-a mRNA, IL-lpmRNA, IL-8mRNA expression with LPS stimulation were investigated by RT-PCR technique; ELIS A technique measured TNF-a, IL-6 contents of J774 cells, neutrophil chemoattractant made with zymosan was observed.Results The results from RT-PCR showed that TSDA could inhibit EL-1 pmRNA,IL-8mRNA expression, the inhibitory rates on IL-lp*mRNA expression in the groups of 100, 1000 mg.L"1 were about 27.2±7.8%> 32.5±10.8% (P<0.01,n=4) respectively; the inhibitory rates on IL-8mRNA expression in the groups of 100,1000 mg.L"1 were about 19.1±7.2%^ 24.0±6.2% (P<0.05,n=4) respectively. In addition, TSDA 10, 100, 1000 mg-L^could inhibit TNFa and IL-6 secretion of J774 cells, the inhibitory rates on TNFa were 32.8%, 56.6%, 85.6% and on IL-6 were 33.4%, 61.8%, 86.1%. And TSDA 1000 mg.L"1 could inhibit TNFamRNA expression, the inhibitory rate was 15.0±6.0%. What's more TSDA 100, 1000 mg.L"1 could inhibit neutrophil chemotaxis induced by zymosan.Conclusion TSDA could inhibit the synthesis of TNF-a , IL-6 and expression of IL-ipmRNA, IL-8mRNA,TNF-amRNA. Inhibition inflammatory cytokines production
目的 从牛膝、萆薢中分别提取总皂苷,测定提取物中总皂苷含量,了解提取物用于药效学试验的可行性。
方法 牛膝切片采用水提、石油醚脱脂、醇提、饱和正丁醇萃取、过氧化铝柱提取总皂苷,采用紫外—可见分光光度法,以齐墩果酸为对照品测定牛膝提取物中总皂苷的含量。萆薢切片乙醇提取、石油醚脱脂、饱和正丁醇萃取、加丙酮沉淀提取总皂苷,采用紫外—可见分光光度法,以薯蓣皂苷元为对照品测定测定萆薢提取物中总皂苷的含量。
结果 齐墩果酸在2.77 μg～13.85 μg范围内线性关系良好,平均加样回收率为97.0%,牛膝提取物中总皂苷含量为80.%。薯蓣皂苷元在5.12 μg～25.6 μg范围内线性关系良好,样品平均加样回收率为97.5%。萆薢提取物中总皂苷含量为53.1%。
结论 上述提取总皂苷方法可行,稳定,总成分含量能满足药效学试验的要求。
第二部分 萆薢总皂苷+牛膝总皂苷的组方合理性研究
目的 从降低血尿酸水平和抗炎作用两方面探讨牛膝总皂苷和萆薢总皂苷的组方合理性及剂量配比。
方法 采用尿酸型小鼠高尿酸血症模型和酵母膏致小鼠高尿酸血症模型以及小鼠耳肿胀模型和微晶型尿酸钠致大鼠急性痛风性关节炎模型模型。按照Codrug软件,得优化组方。
结果 在尿酸型小鼠高尿酸血症模型和酵母膏致小鼠高尿酸血症模型,萆薢总皂苷组、牛膝总皂苷+萆薢总皂苷组和阳性对照组小鼠血清尿酸水平显著降低(P<0.01),牛膝总皂苷480、240、120 mg.kg~(-1)组小鼠血清尿酸水平与模型组比较无显著性差异。萆薢总皂苷组与牛膝总皂苷+萆薢总皂苷组相比小鼠血清尿酸水平也无明显差异。牛膝总皂苷、萆薢总皂苷对小鼠耳肿胀均有显著的抑制作用,牛膝总皂苷+萆薢总皂苷组耳肿胀抑制率均强于单药组。
Part 1 Extracting of total saponin and research on their qualityAim To extract the total saponin of Dioscorea and Achyranthes and research their quality. Methods The total saponin of Achyranthes bidentata (TSA) was extracted by water, alcohol, n-butanol.With oleanolic acid as control, the content of total saponin was determinated by UV. Total saponin of Dioscorea (TSD) was extracted by alcohol, n-butanol. With diosgenin as control, the content of total saponin was determinated by UV.Results The linear relationship of oleanolic acid was better between 2.77μg~13.85μg, average recovery rate was 97% and the content of total saponin of Achyranthes was 80%. The linear relationship of diosgenin was better between 5.12μg~25.6μg, average recovery rate was 97.5% and the content of total saponin was Dioscorea 53.1%.Conclusion The methods was well done to extract total saponin and stable. The ingrements comformed to the rules of animal experiments.Part 2 Study on the rationality of TSD plus TSAAim To study the rationality and the dose of TSD and TSA according to the decrease ofserum uric acid level and anti-inflammatory of TSD and TSA.Methods Mice hyperuricemic models were made by uric acid ip or yeast extract paste ig. Inthese models the effect of TSD, TDA on uric acid level was observed. And mice ear swellingwas induced by dimethylbenzene, rat feet swelling was induced by MSU. In these modelsanti-inflammatory effect of TSD, TSA was measured. According to Codrug software, thegrading-up combination prescription was obtained.Results In the mice hyperuricemic models, the serum uric acid level of TSD group, TSDplus TSA group and positive control group was significantly reduced compared with the
model group(P < 0. 01) .The serum uric acid level of TSA group has no difference from that of the model group. The serum uric acid level of TSD group has no difference from that TSD plus TSA group yet. Both of TSD and TSA could markedly inhibit the mice ear swelling. Moreover the inhibitory of the group of TSD plus TSA was more stronger than the group of TSD or TSA. In the mice ear swelling model, TSA120 mg-kg'1 plus TSD 240mg. kg"1 could reach the maximal inhibitory effect; in the rat feet swelling model TSA 60 mg-kg"1 plus TSD 120 mg. kg"1 could reach the maximal inhibitory effect.Conclusion TSD could reduce the mice serum uric acid level, but TSA could not. Both TSD and TSA had anti-inflammatory effect. TSD plus TSA had more anti-inflammatory effect. In these models the best proportion of TSA and TSD was 1:2.Part 3 Establish animal hyperuricemia model and studies on the effects and mechanisms of TSD A on animal experimental hyperuricemiaAim To establish animal hyperuricemia model and studies on the effects and mechanisms of TSDA on animal experimental hyperuricemia.Methods (l)Mouse was given different dose of uric acid by intraperitoneal injection(ip), the serum uric acid level was detected after 1 h later. (2) Yeast extract paste 7. 5—30 g ? kg"1 was given to mice by ig once daily for 7,14,21,28 consecutive days, and detected the level of uric acid in serum.(3)Mouse and rat hyperuricemic models were made by orally administering yeast extract paste once per day (30g.kg'], 20g.kg"', respectively) for 7 days; another mouse hyperuricemia model was generated by intraperitoneal injection once with uric acid 250 mg.kg"1 or potassium oxonate 300 mg-kg"1. (4)The concentration of uric acid in serum or urine was detected by the phosphotungstic acid method, and the activity of xanthine oxidase (XOD) was assayed by a test kit.Results (1) Uric acid 125, 250, 500, 1000 mg.kg1 ip could significantly increase the level of uric acid in mouse serum. The level of uric acid in mouse serum was attained peak at 10
min and the hyperuricemia could lasted over 4 hours after uric acid 250mg.kg"Igiven. (2)The serum uric acid level in mice were (271. 8±53. 2), (215. 4+31. 5), (195. 9 + 56. 0), (142.1 ±30. 7) umol ? L"1 in 30 g - kg"1 group; and (226. 8±40. 7), (148. 67±30. 4), (176. 9+27.0), (119.3±27. 4) umol . L"1 in 15 g . kg'1 group after administration 1, 2, 3, 4 weeks; In 7. 5 g ? kg'1 group the serum uric acid level was (117. 0+29.0) umol ? L"1 after 1 week. However the serum uric acid of the control group was (108.1 ± 13.9) umol ? L"1. (3)In the mouse hyperuricemia model induced by uric acid, the serum uric acid of the normal group, model group m,TSD plus TSA group (400 ^ 120n 40 mg. kg"1), benzbromarone group (20 mg. kg"1) and allopurinol group (40 mg. kg"1) were 114. 3+21. 4, 229.6+46.9, 160.4 + 32.0, 162.5 + 27.5, 25. 4 ±35. 5, 126.9 ±19.8, 189. 6 ± 31. 6 u mol. L1 respectively.(4) In the mouse hyperuricemia model induced by yeast extract paste, the serum uric acid level were 118.6±29.2, 244.7±47.2, 161.7±34.6, 144.8±31.0, 175.6± 23.9, 81.8±20. 7, 73.0±19.4umol.L~1 respectively. (5) In the mouse hyperuricemia model induced by potassium oxonate, the serum uric acid level were 98. 2+29.9,179. 7± 39.0,131. 7 ± 20.0,140.8 ± 29.2,160. 6 ± 28. 2,88. 7 ± 21.2,117. 3 ± 32.3 u mol. L"1 respectively. (6) In the rat hyperuricemia model induced by yeast extract paste, the serum uric acid level were 152.6+27.9, 282.2±65. 2, 195. 3±66.3, 190. 7±35.8, 243.8+51.1, 178.6 ±40. 7, 121.3 ±57.3 umol.L"1 respectively; the serum XOD activity were 33. 6±3. 2, 52. 4±4. 4, 42. 3±5. 9, 44. 9±5. 1, 44. 6±6. 7, 48. 8±7. 0, 10.8+2. 2 U. I/1 respectively; 24 h total uric acid excretion, were 23. 5 ±4.82, 43.1 +9.18, 41. 4 ±8.53, 35.3±6. 7, 36. 7±8. 06, 43.8±12. 6, 19. 2±4. 9 mmol respectively.Conclusion (1) Uric acid given by ip could form mice hyperuricemia model, the dosage of 250 mg .kg"1 was better. (2) Yeast extract paste (15~30) g ? kg"1 given by ig could form mice hyperuricemia model, and the hyperuricemia could last 1 ~2 weeks.(3) TSDA possesses a potent anti-hyperuricemic effect on hyperuricemic animals, and the mechanism may be relevant in accelerating the excretion and decreasing the production of uric acid. Part 4 Eeffects and mechanisms of TSDA on gouty arthritis of
rabbit and ratAim To observe the preventive and therapeutic effects and mechanisms of TSD on animal gout arthritis.Methods Rat gouty arthritis model and rabbit gouty arthritis model were established by MSU intraarticular injection, the joint swell degree, inflammatory factor and cytokine in the joint fluid or leachate were detected, moreover histopathologic examination was done, rat feet swelling was induced by carrageenan, mice ear swelling was induced by dimethylbenzene, the mice writhing responewas induced by acetic acid, the mice pain induced by warm bath or hot plate, and rat granuloma induced by cotton ball subcutaneously embedding. In these models the swelling degree, pain response latency, granuloma weight and so on were detected respectively.Results hi the rat model, the joint swelling was very obviouse 6 h later and the swelling degree reached the peak at 24h or 48h.The level of NO> TNF-ou PGE2. IL-lp\ IL-6 and IL-8 in the model group was significantly increased and histopathologic examination showed that the synovial cells were seriously injured in the model group, there are many polycyte, histoleucocyte and urete crystal in the joint and soft tissue. TSDA 200, 60mg.kg'1 could significantly inhibit joint swelling of rat and significantly improve the histopathologic changes. TSDA 200mg.kg"1 could improve the gaits and significantly inhibit the increase of NO. TNF- a and PGE2 content as well as the content of IL-6.In the rabbit model, the rabbit joint of the model group appeared red, swelling and higher skin temperature; the fluid increased; WBC in the blood and fluid rose significantly in the model group. The level of NCK TNF-ou PGE2> IL-lp\ IL-6 and IL-8 in the model group was significantly increased and histopathologic examination showed that the synovial cells and soft tissue were seriously injured in the model group. TSDA 100 mg.kg"1 could significantly reduce the fluid in the joint and 100, 30, 10 mg. kg"1 could significantly reduce WBC in the joint fluid. 100, 30 mg.kg"1 inhibited the content of IL-lfi. IL-6 and IL-8.In the model induced by carrageenan, the swellling degree of TSDA 200,60mg.kg" groups was significantly decreased compared with the model group. In the model of cotton
ball granuloma, the weight of granuloma groups were significantly reduced. In the modelsinduced by dimethylbenzen and writhing respone was induced by acetic acid, TSDA400,120,40 mg.kg'1 could significantly reduce mouse ear swelling and frequency of writhingrespone. TSDA 400 mg.kg"! could prolong pain response latency.Conclusion TSDA possessed antigout arthritis effects, its mechanism may be related toinhibit the product of cytokines.Part 5 Effects of TSDA on cytokine secrection and their gene expression of J774 cells and human peripheral blood mononuclear cellsAim To investigate the effects of TSDA on cytokine secrection and their gene expression of J774 cells and human peripheral blood mononuclear cells (PBMC) in vitro. The effect of TSDA on neutrophil chemotaxis was also observed.Methods Isolated total RNA from different groups of J774 cells and PBMC, TNF-a mRNA, IL-lpmRNA, IL-8mRNA expression with LPS stimulation were investigated by RT-PCR technique; ELIS A technique measured TNF-a, IL-6 contents of J774 cells, neutrophil chemoattractant made with zymosan was observed.Results The results from RT-PCR showed that TSDA could inhibit EL-1 pmRNA,IL-8mRNA expression, the inhibitory rates on IL-lp*mRNA expression in the groups of 100, 1000 mg.L"1 were about 27.2±7.8%> 32.5±10.8% (P<0.01,n=4) respectively; the inhibitory rates on IL-8mRNA expression in the groups of 100,1000 mg.L"1 were about 19.1±7.2%^ 24.0±6.2% (P<0.05,n=4) respectively. In addition, TSDA 10, 100, 1000 mg-L^could inhibit TNFa and IL-6 secretion of J774 cells, the inhibitory rates on TNFa were 32.8%, 56.6%, 85.6% and on IL-6 were 33.4%, 61.8%, 86.1%. And TSDA 1000 mg.L"1 could inhibit TNFamRNA expression, the inhibitory rate was 15.0±6.0%. What's more TSDA 100, 1000 mg.L"1 could inhibit neutrophil chemotaxis induced by zymosan.Conclusion TSDA could inhibit the synthesis of TNF-a , IL-6 and expression of IL-ipmRNA, IL-8mRNA,TNF-amRNA. Inhibition inflammatory cytokines production
引文
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2. Kamienski M. Gout: not just for the rich and famous! Everyman's disease. Orthop Nurs, 2003; 22(1): 16～20
3. Arromdee E, Michet C J, Crowson CS, et al. Epidemiology of gout: is the incidence rising? JRheumatol, 2002; 29(11): 2403～6
4. Yu KH, Luo SF. Younger age of onset of gout in Taiwan. Rheumatology (Oxford), 2003; 42(1): 166～70
5. Kuzuya M, Ando F, Iguchi A, Shimokata H. Effect of aging on serum uric acid levels: longitudinal changes in a large Japanese population group. J Gerontol A Biol Sci Med Sci, 2002; 57(10): M660～4
6. Kim KY, Ralph Schumacher H, Hunsche E, et al. A literature review of the epidemiology and treatment of acute gout. Clin Ther, 2003; 25(6): 1593～617
7. Chen S, Du H, Wang Y, Xu L. The cpidemiology study of hyperuricemia and gout in a community population of Huangpu district in Shanghai. Chin Med J, 1998, 111(3): 228
8. Zhu Shi, Tang Ping, Xie Ling, et al. Epidemiological survey(1999-2000) on cardiovascular risk factors in Chengdu hyperuric acid and clinical implication. Chin JHypertens, 2002; 10(5): 476～8
9.邵继红,莫宝庆,喻荣彬,等.南京市社区人群高尿酸血症与痛风的流行病学调查.疾病控制杂志,2003;7(4):305～8
10.申延宏,陈南衡,高嘉琳.老年糖尿病无症状高尿酸血症的临床分析.实用老年医学,1997;11(6):259～60
11.陈慎仁,彭璇娟,蔡应本.原发性高尿酸血症及其相关疾病的调查.临床内科杂志,1997;14(1):42～3
12.李东,曹丽华,张惠英等.高尿酸血症370例分析.中华风湿病学杂志,1999;3(4):244～6
13.杨岫岩,唐福林,尹培达.21家医院痛风住院构成比15年变化趋势分析.中华流行病学杂志,1996;17(1):10～2
14. Chang HY, Pan WH, Yeh WT, Tsai KS. Hyperuricemia and gout in Taiwan: results from the nutritional and health survey in Taiwan (1993-96). J Rheumatol, 2001; 28(7): 1640
15. Ko YC, Wang TN, Tsai LY, et al. High prevalence of hyperuricemia in adolescent Taiwan aborigines. J Rheumatol, 2002; 29(4): 837～42
16.张振文.痛风的危险因素.国外医学内分泌分册,1997;17(3):141～3
17.华鸿宝,殷安康,蔡九钟等.抗高尿酸血症追踪治疗对原发性痛风的影响.中华内分泌代谢杂志,1996;12(3):135～7
18. Punzi L, Calo L, Plebani M. Clinical significance of cytokine determination in synovial fluid. Crit Rev Clin Lab Sci, 2002; 39(1): 63～88
19.庄宏.高尿酸血症及老年人原发性痛风的研究.中华老年医学杂志,1993;12(5):314～6
20. Koh WH, Seah A, Chai P. Clinical presentation and disease associations of gout: a hospital based study of 100 patients in Singapore. Ann Acad Med Singapore. 1998, 27(1): 7～10
21. Takahashi S, Yamanoto T, Moriwaki Y et al. Increased eoneentration of serum Lp (a) lipoprotein in patients with primary gout. Ann Rheum Dis, 1995; 54(2): 90～3
22. Tomita M, Mizuno S, Yamanaka H, Hosoda Y. Does hyperuricemia affect mortality? A prospective cohort study of Japanese male workers. J Epidemiol, 2000; 10(6): 403～9
23.Jing Fang,Alderman MH.血清尿酸与心血管病死亡率:1971-1992年首次国民健康与营养状况普查及流行病学随访研究.美国医学会杂志·中文版,2001;20(2):53～9
24.陈光亮,徐叔云.高尿酸血症研究进展.中国药理学通报,2003;19(10):1088～92
25.胡俊发,郭素缄,陈树明.110例高尿酸血症及痛风临床观察.天津医药,1996,24(6):353～5
26. Grwitz JH Kalish SC, Bohn RL et al. Thiazide diuretics and the initiation of anti-gout therapy, J Clin Epidemiol, 1997; 50 (8): 953～9
27.田国庆.中医辨证及治疗痛风研究近况.医学研究通讯,1995;24(4):17～19
28. Pollmarm G, Kullich W, Klein G. Therapy of hyperuricemia and gout. Wien Med Wochenschr, 1997; 147 (16): 382～7
29. Emmerson BT. The management of gout. NEngldMed, 1996; 334(9): 445～51
30.陈光亮,王琳琳,徐叔云.防治痛风的药物研究进展.国外医学内分泌学分册,2005年,排印中
31. Fam AG. Managing problem gout. AnnAcadMedSingapore. 1998; 27 (1): 93～9
32. Shimizu T. Treatment for gout arthitis. Nippon Rinsho, 1996; 54(12): 3267～71
33. Tamanaka H. How to select and use urate lowering for hyperuricemia, Nippon Rinsho, 1996; 54(12): 3261～5
34. Conaghan PG, Day RO. Risk and benefits of drugs used in the management and prevention of gout. Drugs, 1994; 11(4): 252～8
35. Ben Chetrit E, Levy M. Colchicine: 1998 update. Semin Arthritis Rheum, 1998; 28 (1): 48～59
36. Martinez RV, Reval M, Campos MD, et al. Involvement of peripheral eyelooxygenase-1 and eyclooxygenase-2 in inflammatory pain. JPharm Pharmacol, 2002; 54(3): 405
37.纪元.警告:抗痛风药苯溴马隆可致重症肝炎.日本医学介绍,2000;21(6):285
38. Yamada H, Kotaki H, Furitsu H, et al. Mechanism of the uricosuric action of E3040, a drug used to treat inflammatory bowel disease Ⅱ: study using DBA. 2N mice. Biopharm Drug Dispos, 1999; 20(5): 271～6
39. Sumino H, Iehikawa S, Kanda T, et al. Reduction of serum uric acid by hormone replacement therapy in postmenopausal women with hyperuricaemia. Lancet, 1999; 354(9179): 650～2
40. Mukhin IV, Ignatenko GA, Nikolenko VY. Dyshormonal disorders in gout: experimental and clinical studies. Bull Exp Biol Med, 2002; 133(5): 491～3
41.陈光亮,徐叔云.中药治疗痛风研究近况.安徽中医学院学报,2003;22(5):57～9
42.陈文照,姜宏,顾瑞生等.经方治疗痛风临床研究进展.中医正骨,1996;8(2):33
43.张梅红,周翠英.痛风的中医药研究进展.山东中医学院学报,1996;20(6):421
44.陈光亮,王钦茂,徐叔云.小鼠高尿酸血症模型的研究.中国药理学通报,2001;17(3):351
45.陈光亮,张清林,徐叔云,等.酵母致小鼠高尿酸血症模型.中国药理学通报,2003;19(4):467～9
46.陈光亮,徐叔云.高尿酸血症动物模型研究进展.中国药理学通报,2004;20(4):369～73
47.陈云凤,扈晓宇,彭介寿.痛风宁Ⅰ号抗炎及镇痛作用.中药药理与临床,2000,16(4):38
48.陈文照,金策,林坚等.痛风宁对尿酸钠致大鼠关节炎模型前列腺素的影响.中 国医药学报,2000;15(2):104～6
49.林学波.中医中药治疗急性痛风性关节炎.广东医学,1999;20(10):813～4
50.张玉红.三妙汤加味治疗痛风急发67例.云南中医中药杂志,2000:21(1):21～2
51.许少素.加味四妙汤治疗老年痛风性关节炎30例.安徽中医临床杂志,1999;11(6):397
52.陈文照,姜宏,陈家荣.痛风宁治疗急性痛风55例临床观察.中医正骨,1999;11(11):25～6
53.王纪云,谢梅,甄艳,等.痛风散合四妙汤加味治疗急性痛风性关节炎42例.云南中医中药杂志.1999;20(2):14～5
54.许伏新,侯士良.三妙散镇痛抗炎作用的实验研究.基层中药杂志.1999;13(1):15～7
55.郝钰,沈欣,邱全瑛.情热燥湿法对类风湿性关节炎治疗作用的实验研究.中国中医基础医学杂志,1997:3(1):32～34
56.李勤凡,童德文,朱小甫,等.萆薢提取物治疗鸡痛风的剂量筛选试验.西北农林科技大学学报(自然科学版),2001;29(3):84～86
57.童德文,耿果霞,赵宝玉,等.萆薢中薯蓣皂甙分离及其对鸡痛风的作用.西北农林大学学报,1999:27(6):108～111
58.国家中医药管理局《中华本草》编委会.《中华本草》(精选本,上、下册),第1版,上海:上海科技出版社出版,1998;402～9,2107～11,585～91,1039～50,1885～93,2139～44
59.王浴生,邓文龙,薛春生主编.中药药理与临床.第2版,北京:人民卫生出版社,1998:480～95,1024～9
60.张前德.萆薢丸加味治疗痛风性关节炎26例.新中医,2000;32(10):47
61.陈国新.萆薢分清饮治疗痛风性关节炎.陕西中医,1999:20(12):564～5
62.王清伟.萆薢、土茯苓、菝葜的临床鉴别及应用.陕西中医,2000:21(8):378
63.赵维良.浙江产薯蓣属萆薢组药材的化学成分综述.现代应用药学,1988;(1):38～40
64.郑宝灿,陈忠科.怀牛膝多倍体、单体和二倍体的药理作用比较.药学通报,1988;23(11):666
65.史玉芬,郑延彬.牛膝抗炎、抗菌作用的研究.中药通报,1988;13(7):44
2. Kamienski M. Gout: not just for the rich and famous! Everyman's disease. Orthop Nurs, 2003; 22(1): 16～20
3. Arromdee E, Michet C J, Crowson CS, et al. Epidemiology of gout: is the incidence rising? JRheumatol, 2002; 29(11): 2403～6
4. Yu KH, Luo SF. Younger age of onset of gout in Taiwan. Rheumatology (Oxford), 2003; 42(1): 166～70
5. Kuzuya M, Ando F, Iguchi A, Shimokata H. Effect of aging on serum uric acid levels: longitudinal changes in a large Japanese population group. J Gerontol A Biol Sci Med Sci, 2002; 57(10): M660～4
6. Kim KY, Ralph Schumacher H, Hunsche E, et al. A literature review of the epidemiology and treatment of acute gout. Clin Ther, 2003; 25(6): 1593～617
7. Chen S, Du H, Wang Y, Xu L. The cpidemiology study of hyperuricemia and gout in a community population of Huangpu district in Shanghai. Chin Med J, 1998, 111(3): 228
8. Zhu Shi, Tang Ping, Xie Ling, et al. Epidemiological survey(1999-2000) on cardiovascular risk factors in Chengdu hyperuric acid and clinical implication. Chin JHypertens, 2002; 10(5): 476～8
9.邵继红,莫宝庆,喻荣彬,等.南京市社区人群高尿酸血症与痛风的流行病学调查.疾病控制杂志,2003;7(4):305～8
10.申延宏,陈南衡,高嘉琳.老年糖尿病无症状高尿酸血症的临床分析.实用老年医学,1997;11(6):259～60
11.陈慎仁,彭璇娟,蔡应本.原发性高尿酸血症及其相关疾病的调查.临床内科杂志,1997;14(1):42～3
12.李东,曹丽华,张惠英等.高尿酸血症370例分析.中华风湿病学杂志,1999;3(4):244～6
13.杨岫岩,唐福林,尹培达.21家医院痛风住院构成比15年变化趋势分析.中华流行病学杂志,1996;17(1):10～2
14. Chang HY, Pan WH, Yeh WT, Tsai KS. Hyperuricemia and gout in Taiwan: results from the nutritional and health survey in Taiwan (1993-96). J Rheumatol, 2001; 28(7): 1640
15. Ko YC, Wang TN, Tsai LY, et al. High prevalence of hyperuricemia in adolescent Taiwan aborigines. J Rheumatol, 2002; 29(4): 837～42
16.张振文.痛风的危险因素.国外医学内分泌分册,1997;17(3):141～3
17.华鸿宝,殷安康,蔡九钟等.抗高尿酸血症追踪治疗对原发性痛风的影响.中华内分泌代谢杂志,1996;12(3):135～7
18. Punzi L, Calo L, Plebani M. Clinical significance of cytokine determination in synovial fluid. Crit Rev Clin Lab Sci, 2002; 39(1): 63～88
19.庄宏.高尿酸血症及老年人原发性痛风的研究.中华老年医学杂志,1993;12(5):314～6
20. Koh WH, Seah A, Chai P. Clinical presentation and disease associations of gout: a hospital based study of 100 patients in Singapore. Ann Acad Med Singapore. 1998, 27(1): 7～10
21. Takahashi S, Yamanoto T, Moriwaki Y et al. Increased eoneentration of serum Lp (a) lipoprotein in patients with primary gout. Ann Rheum Dis, 1995; 54(2): 90～3
22. Tomita M, Mizuno S, Yamanaka H, Hosoda Y. Does hyperuricemia affect mortality? A prospective cohort study of Japanese male workers. J Epidemiol, 2000; 10(6): 403～9
23.Jing Fang,Alderman MH.血清尿酸与心血管病死亡率:1971-1992年首次国民健康与营养状况普查及流行病学随访研究.美国医学会杂志·中文版,2001;20(2):53～9
24.陈光亮,徐叔云.高尿酸血症研究进展.中国药理学通报,2003;19(10):1088～92
25.胡俊发,郭素缄,陈树明.110例高尿酸血症及痛风临床观察.天津医药,1996,24(6):353～5
26. Grwitz JH Kalish SC, Bohn RL et al. Thiazide diuretics and the initiation of anti-gout therapy, J Clin Epidemiol, 1997; 50 (8): 953～9
27.田国庆.中医辨证及治疗痛风研究近况.医学研究通讯,1995;24(4):17～19
28. Pollmarm G, Kullich W, Klein G. Therapy of hyperuricemia and gout. Wien Med Wochenschr, 1997; 147 (16): 382～7
29. Emmerson BT. The management of gout. NEngldMed, 1996; 334(9): 445～51
30.陈光亮,王琳琳,徐叔云.防治痛风的药物研究进展.国外医学内分泌学分册,2005年,排印中
31. Fam AG. Managing problem gout. AnnAcadMedSingapore. 1998; 27 (1): 93～9
32. Shimizu T. Treatment for gout arthitis. Nippon Rinsho, 1996; 54(12): 3267～71
33. Tamanaka H. How to select and use urate lowering for hyperuricemia, Nippon Rinsho, 1996; 54(12): 3261～5
34. Conaghan PG, Day RO. Risk and benefits of drugs used in the management and prevention of gout. Drugs, 1994; 11(4): 252～8
35. Ben Chetrit E, Levy M. Colchicine: 1998 update. Semin Arthritis Rheum, 1998; 28 (1): 48～59
36. Martinez RV, Reval M, Campos MD, et al. Involvement of peripheral eyelooxygenase-1 and eyclooxygenase-2 in inflammatory pain. JPharm Pharmacol, 2002; 54(3): 405
37.纪元.警告:抗痛风药苯溴马隆可致重症肝炎.日本医学介绍,2000;21(6):285
38. Yamada H, Kotaki H, Furitsu H, et al. Mechanism of the uricosuric action of E3040, a drug used to treat inflammatory bowel disease Ⅱ: study using DBA. 2N mice. Biopharm Drug Dispos, 1999; 20(5): 271～6
39. Sumino H, Iehikawa S, Kanda T, et al. Reduction of serum uric acid by hormone replacement therapy in postmenopausal women with hyperuricaemia. Lancet, 1999; 354(9179): 650～2
40. Mukhin IV, Ignatenko GA, Nikolenko VY. Dyshormonal disorders in gout: experimental and clinical studies. Bull Exp Biol Med, 2002; 133(5): 491～3
41.陈光亮,徐叔云.中药治疗痛风研究近况.安徽中医学院学报,2003;22(5):57～9
42.陈文照,姜宏,顾瑞生等.经方治疗痛风临床研究进展.中医正骨,1996;8(2):33
43.张梅红,周翠英.痛风的中医药研究进展.山东中医学院学报,1996;20(6):421
44.陈光亮,王钦茂,徐叔云.小鼠高尿酸血症模型的研究.中国药理学通报,2001;17(3):351
45.陈光亮,张清林,徐叔云,等.酵母致小鼠高尿酸血症模型.中国药理学通报,2003;19(4):467～9
46.陈光亮,徐叔云.高尿酸血症动物模型研究进展.中国药理学通报,2004;20(4):369～73
47.陈云凤,扈晓宇,彭介寿.痛风宁Ⅰ号抗炎及镇痛作用.中药药理与临床,2000,16(4):38
48.陈文照,金策,林坚等.痛风宁对尿酸钠致大鼠关节炎模型前列腺素的影响.中 国医药学报,2000;15(2):104～6
49.林学波.中医中药治疗急性痛风性关节炎.广东医学,1999;20(10):813～4
50.张玉红.三妙汤加味治疗痛风急发67例.云南中医中药杂志,2000:21(1):21～2
51.许少素.加味四妙汤治疗老年痛风性关节炎30例.安徽中医临床杂志,1999;11(6):397
52.陈文照,姜宏,陈家荣.痛风宁治疗急性痛风55例临床观察.中医正骨,1999;11(11):25～6
53.王纪云,谢梅,甄艳,等.痛风散合四妙汤加味治疗急性痛风性关节炎42例.云南中医中药杂志.1999;20(2):14～5
54.许伏新,侯士良.三妙散镇痛抗炎作用的实验研究.基层中药杂志.1999;13(1):15～7
55.郝钰,沈欣,邱全瑛.情热燥湿法对类风湿性关节炎治疗作用的实验研究.中国中医基础医学杂志,1997:3(1):32～34
56.李勤凡,童德文,朱小甫,等.萆薢提取物治疗鸡痛风的剂量筛选试验.西北农林科技大学学报(自然科学版),2001;29(3):84～86
57.童德文,耿果霞,赵宝玉,等.萆薢中薯蓣皂甙分离及其对鸡痛风的作用.西北农林大学学报,1999:27(6):108～111
58.国家中医药管理局《中华本草》编委会.《中华本草》(精选本,上、下册),第1版,上海:上海科技出版社出版,1998;402～9,2107～11,585～91,1039～50,1885～93,2139～44
59.王浴生,邓文龙,薛春生主编.中药药理与临床.第2版,北京:人民卫生出版社,1998:480～95,1024～9
60.张前德.萆薢丸加味治疗痛风性关节炎26例.新中医,2000;32(10):47
61.陈国新.萆薢分清饮治疗痛风性关节炎.陕西中医,1999:20(12):564～5
62.王清伟.萆薢、土茯苓、菝葜的临床鉴别及应用.陕西中医,2000:21(8):378
63.赵维良.浙江产薯蓣属萆薢组药材的化学成分综述.现代应用药学,1988;(1):38～40
64.郑宝灿,陈忠科.怀牛膝多倍体、单体和二倍体的药理作用比较.药学通报,1988;23(11):666
65.史玉芬,郑延彬.牛膝抗炎、抗菌作用的研究.中药通报,1988;13(7):44