六味痛风饮防治痛风性关节炎作用机理的实验研究
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摘要
本研究提出了“痛风病位在肝,与脾肾密切相关;肝脾肾功能失调为本,肝经浊毒、湿热蕴蒸为其关键致病因素”的创新观点,精选了临床行之有效的药物自拟六味痛风饮以清肝泄浊、补脾益肾,标本兼治。为进一步评估其疗效,建立急性痛风性关节炎大鼠及高尿酸血症鹌鹑模型,进行实验研究。
目的:探讨六味痛风饮治疗痛风性关节炎及预防其复发的作用机理。
方法:(1)急性痛风性关节炎实验研究:应用Coderre经典的造模方法造模,分别予六味痛风饮、秋水仙碱、醋氯芬酸进行灌胃治疗,观察大鼠受试关节步态、关节肿胀周径及组织病理学的变化。采用酶联免疫(ELISA)法,检测各组大鼠受试关节内MMP-3、IFN-γ、IL-4的含量;同时观察各治疗组对小鼠扭体实验的镇痛作用。(2)高尿酸血症的实验研究:采用给鹌鹑灌腺嘌呤的方法造模,分别给予六味痛风饮、别嘌呤醇、苯溴马隆进行灌胃,观察鹌鹑血尿酸及黄嘌呤氧化酶的含量变化。
结果:(1)六味痛风饮、秋水仙碱及醋氯芬酸均明显改善模型大鼠的步态(P<0.05)。(2)六味痛风饮、秋水仙碱、醋氯芬酸能抑制急性痛风性关节炎大鼠关节肿胀,各治疗组关节周径与模型组相比有差异(P<0.05),各治疗组间无差异(P>0.05)。(3)病理观察显示,六味痛风饮、秋水仙碱及醋氯芬酸能改善痛风性关节炎模型关节滑膜炎症病理改变。(4)关节周围组织MMP-3含量在空白组、各治疗组均低于模型组,其差异有意义(P<0.05);各治疗组之间无差异(P>0.05)。(5)关节周围组织IFN-γ含量在空白组、各治疗组均低于模型组,其差异有意义(P<0.05),各治疗组之间无差异(P>0.05)。(6)关节周围组织IL-4含量在空白组、各治疗组均高于模型组,差异无统计学意义(P>0.05)。(7)各治疗组均能减轻醋酸引起的小鼠腹部疼痛,与对照组比较,均有显著性差异(P<0.01),各用药组之间相比,没有差异(P>0.05)。(8)高尿酸血症鹌鹑模型造模成功,造模组血尿酸明显高于空白组,其差异具有显著意义(P<0.01)。(9)血尿酸在六味痛风饮组、别嘌呤醇组及苯溴马隆组较用药前明显下降,其差异有意义(分别为P<0.05,P<0.01,P<0.01);药物治疗后,三组血尿酸低于模型组,其差异有意义(分别为P<0.05,P<0.01,P<0.01);而各治疗组之间两两比较,无明显差异(均为P>0.05)。(10)血XOD在六味痛风饮组、别嘌呤醇组较用药前明显下降,其差异具有统计学意义(分别为P<0.05,P<0.01);药物治疗后,两组血XOD均低于模型组,其差异有意义(分别为P<0.05,P<0.01)。苯溴马隆组血XOD较用药前下降,其差异无意义(P>0.05);治疗后,苯溴马隆组与模型组比较,无统计学意义(P>0.05)。六味痛风饮组血XOD高于别嘌呤醇组,低于苯溴马隆组,差异均有统计学意义(P<0.05);别嘌呤醇组血XOD明显低于苯溴马隆组,有显著差异(P<0.01)。
结论:(1)六味痛风饮对痛风性关节炎大鼠模型具有良好的抗炎消肿及镇痛作用。(2)痛风性关节炎模型关节周围组织有MMP-3、IFN-γ的表达,未见IL-4的明显表达。(3)六味痛风饮可以通过抑制痛风性关节炎模型关节周围组织的MMP-3、IFN-γ的活性,减轻关节周围组织的炎症反应,可能与IL-4的表达无关。(4)成功制作鹌鹑高尿酸血症模型,血尿酸及黄嘌呤氧化酶升高。(5)六味痛风饮降低了高尿酸血症鹌鹑模型的血尿酸、血清黄嘌呤氧化酶活性。(6)六味痛风饮可以有效预防及治疗痛风性关节炎。
In this study, the innovative opinion was that "gout disease in the liver, spleen and kidney dysfunction in liver-based, liver toxicity by muddy, Yun steaming hot and humid are its key pathogenic factor". Liu wei tongfeng-Drink was selected clinical effective drugs to clear liver discharge muddiness, Tonifying splenorenal, both temporary and permanent cures. In order to further assess its efficacy, we set up in rats with acute gouty arthritis and hyperuricemia quail model, experimental study.
Objective: To explore Liu wei tongfeng-Drink gouty arthritis treatment and prevention of its recurrence mechanism.
Methods: (1) Acute gouty arthritis experimental research: Applications Coderre Ways classic model, made of acute gouty arthritis rat models, respectively Liu wei tongfeng-Drink, colchicine, aceclofenac for gavage treatment, the rats tested joints gait, as well as the circumference of its joint swelling and histopathological changes. Using enzyme-linked immunosorbent assay (ELISA) detection methods, MMP-3, IFN-γ, IL-4 expression changes was observed in rat articular of every group; At the same time, the analgesic effect of the treatment group were observed through the mouse writhing test. (2) Hyperuricemia Experimental studies: application to adenine give quail gavage method, building hyperuricemia quail model, respectively, to Liu wei tongfeng-Drink, allopurinol, benzbromarone for gavage treatment, observation quail serum uric acid and xanthine oxidase Change the content.
Results: (1) Liu wei tongfeng-Drink, colchicine and aceclofenac significantly improve the gait of rats (P<0.05). (2) Liu wei tongfeng-Drink, colchicine, aceclofenac can inhibit acute gouty arthritis joint swelling in rats, joint circumference in the treatment group compared with the model group that has differences (P<0.05), the no difference between the treatment groups (P>0.05). (3) Pathological observation showed that Liu wei tongfeng-Drink, colchicine and aceclofenac can improve the model of gouty arthritis patients the pathological changes of articular synovitis. (4) MMP-3 levels of the joints surrounding tissue in the blank group, the treatment groups were lower than the model group, the difference between meaningful (P<0.05); all is no difference between the treatment groups (P>0.05). (5) IFN-γlevels of the joints surrounding tissue in the blank group, the treatment groups were lower than the model group, the difference between meaningful (P<0.05), between the treatment groups no significant difference (P>0.05). (6) IL-4 content of joints surrounding tissue in the blank group, the treatment groups were higher than model group, the difference meaningless (P>0.05).(7) The treatment groups are able to reduce the acetic acid-induced mouse abdominal pain, compared with the model group, were significantly different (P<0.01), between the treatment groups compared with no difference (P>0.05). (8) Quail model of hyperuricemia model have been successfully set up, model of serum uric acid was significantly higher than the blank group, the difference was significant (P<0.01). (9) Serum uric acid of Liu wei tongfeng-Drink group, Allopurinol and benzbromarone group than it’s group before treatment were significantly decreased, the difference was statistically significant (respectively P<0.05, P <0.01, P<0.01 ); drug treatment, Liu wei tongfeng-Drink group, Allopurinol group and benzbromarone group than those in model group, serum uric acid decreased, the difference was significant (respectively P<0.05, P<0.01, P<0.01); while between the treatment groups compared with no significant differences (all P>0.05). (10) Blood XOD at Liu wei tongfeng-Drink group, Allopurinol group than in their pre-medication has declined markedly, the difference was statistically significant (respectively P<0.05, P<0.01); XOD at Benzbromarone group was lower than its medication before, the fall there was no significant difference (P>0.05); After their drug treatment, blood XOD at Liu wei tongfeng-Drink group, Allopurinol group is lower than those in model group, the difference was statistically significant (P<0.05, P<0.01); blood XOD in benzbromarone group compared with the model group, that was no statistical significance (P>0.05); blood XOD at Liu wei tongfeng-Drink gout than allopurinol group, which is lower than benzbromarone group , the differences were statistically significant (P<0.05); Allopurinol group was significantly lower than blood XOD benzbromarone group, there is significant difference (P<0.01).
Conclusion: (1) Liu wei tongfeng-Drink for gouty arthritis rat model has a good anti-inflammatory and analgesic effect of swelling. (2) Model of gouty arthritis have MMP-3, IFN-γexpression in it’s joint surrounding tissue, no obvious IL-4 expression. (3)Liu wei tongfeng-Drink can reduce surrounding tissue inflammatory response by inhibiting MMP-3, IFN-γactivity of gouty arthritis model articulation surrounding tissue, which may be related to IL-4 expression has nothing to do. (4) The success of the quail hyperuricemia model was set up, their blood uric acid and xanthine oxidase increased. (5) Liu wei tongfeng-Drink can reduce serum uric acid and serum xanthine oxidase activity of hyperuricemia quail model. (6) Liu wei tongfeng-Drink can be effective prevention and treatment of gouty arthritis.
目的:探讨六味痛风饮治疗痛风性关节炎及预防其复发的作用机理。
方法:(1)急性痛风性关节炎实验研究:应用Coderre经典的造模方法造模,分别予六味痛风饮、秋水仙碱、醋氯芬酸进行灌胃治疗,观察大鼠受试关节步态、关节肿胀周径及组织病理学的变化。采用酶联免疫(ELISA)法,检测各组大鼠受试关节内MMP-3、IFN-γ、IL-4的含量;同时观察各治疗组对小鼠扭体实验的镇痛作用。(2)高尿酸血症的实验研究:采用给鹌鹑灌腺嘌呤的方法造模,分别给予六味痛风饮、别嘌呤醇、苯溴马隆进行灌胃,观察鹌鹑血尿酸及黄嘌呤氧化酶的含量变化。
结果:(1)六味痛风饮、秋水仙碱及醋氯芬酸均明显改善模型大鼠的步态(P<0.05)。(2)六味痛风饮、秋水仙碱、醋氯芬酸能抑制急性痛风性关节炎大鼠关节肿胀,各治疗组关节周径与模型组相比有差异(P<0.05),各治疗组间无差异(P>0.05)。(3)病理观察显示,六味痛风饮、秋水仙碱及醋氯芬酸能改善痛风性关节炎模型关节滑膜炎症病理改变。(4)关节周围组织MMP-3含量在空白组、各治疗组均低于模型组,其差异有意义(P<0.05);各治疗组之间无差异(P>0.05)。(5)关节周围组织IFN-γ含量在空白组、各治疗组均低于模型组,其差异有意义(P<0.05),各治疗组之间无差异(P>0.05)。(6)关节周围组织IL-4含量在空白组、各治疗组均高于模型组,差异无统计学意义(P>0.05)。(7)各治疗组均能减轻醋酸引起的小鼠腹部疼痛,与对照组比较,均有显著性差异(P<0.01),各用药组之间相比,没有差异(P>0.05)。(8)高尿酸血症鹌鹑模型造模成功,造模组血尿酸明显高于空白组,其差异具有显著意义(P<0.01)。(9)血尿酸在六味痛风饮组、别嘌呤醇组及苯溴马隆组较用药前明显下降,其差异有意义(分别为P<0.05,P<0.01,P<0.01);药物治疗后,三组血尿酸低于模型组,其差异有意义(分别为P<0.05,P<0.01,P<0.01);而各治疗组之间两两比较,无明显差异(均为P>0.05)。(10)血XOD在六味痛风饮组、别嘌呤醇组较用药前明显下降,其差异具有统计学意义(分别为P<0.05,P<0.01);药物治疗后,两组血XOD均低于模型组,其差异有意义(分别为P<0.05,P<0.01)。苯溴马隆组血XOD较用药前下降,其差异无意义(P>0.05);治疗后,苯溴马隆组与模型组比较,无统计学意义(P>0.05)。六味痛风饮组血XOD高于别嘌呤醇组,低于苯溴马隆组,差异均有统计学意义(P<0.05);别嘌呤醇组血XOD明显低于苯溴马隆组,有显著差异(P<0.01)。
结论:(1)六味痛风饮对痛风性关节炎大鼠模型具有良好的抗炎消肿及镇痛作用。(2)痛风性关节炎模型关节周围组织有MMP-3、IFN-γ的表达,未见IL-4的明显表达。(3)六味痛风饮可以通过抑制痛风性关节炎模型关节周围组织的MMP-3、IFN-γ的活性,减轻关节周围组织的炎症反应,可能与IL-4的表达无关。(4)成功制作鹌鹑高尿酸血症模型,血尿酸及黄嘌呤氧化酶升高。(5)六味痛风饮降低了高尿酸血症鹌鹑模型的血尿酸、血清黄嘌呤氧化酶活性。(6)六味痛风饮可以有效预防及治疗痛风性关节炎。
In this study, the innovative opinion was that "gout disease in the liver, spleen and kidney dysfunction in liver-based, liver toxicity by muddy, Yun steaming hot and humid are its key pathogenic factor". Liu wei tongfeng-Drink was selected clinical effective drugs to clear liver discharge muddiness, Tonifying splenorenal, both temporary and permanent cures. In order to further assess its efficacy, we set up in rats with acute gouty arthritis and hyperuricemia quail model, experimental study.
Objective: To explore Liu wei tongfeng-Drink gouty arthritis treatment and prevention of its recurrence mechanism.
Methods: (1) Acute gouty arthritis experimental research: Applications Coderre Ways classic model, made of acute gouty arthritis rat models, respectively Liu wei tongfeng-Drink, colchicine, aceclofenac for gavage treatment, the rats tested joints gait, as well as the circumference of its joint swelling and histopathological changes. Using enzyme-linked immunosorbent assay (ELISA) detection methods, MMP-3, IFN-γ, IL-4 expression changes was observed in rat articular of every group; At the same time, the analgesic effect of the treatment group were observed through the mouse writhing test. (2) Hyperuricemia Experimental studies: application to adenine give quail gavage method, building hyperuricemia quail model, respectively, to Liu wei tongfeng-Drink, allopurinol, benzbromarone for gavage treatment, observation quail serum uric acid and xanthine oxidase Change the content.
Results: (1) Liu wei tongfeng-Drink, colchicine and aceclofenac significantly improve the gait of rats (P<0.05). (2) Liu wei tongfeng-Drink, colchicine, aceclofenac can inhibit acute gouty arthritis joint swelling in rats, joint circumference in the treatment group compared with the model group that has differences (P<0.05), the no difference between the treatment groups (P>0.05). (3) Pathological observation showed that Liu wei tongfeng-Drink, colchicine and aceclofenac can improve the model of gouty arthritis patients the pathological changes of articular synovitis. (4) MMP-3 levels of the joints surrounding tissue in the blank group, the treatment groups were lower than the model group, the difference between meaningful (P<0.05); all is no difference between the treatment groups (P>0.05). (5) IFN-γlevels of the joints surrounding tissue in the blank group, the treatment groups were lower than the model group, the difference between meaningful (P<0.05), between the treatment groups no significant difference (P>0.05). (6) IL-4 content of joints surrounding tissue in the blank group, the treatment groups were higher than model group, the difference meaningless (P>0.05).(7) The treatment groups are able to reduce the acetic acid-induced mouse abdominal pain, compared with the model group, were significantly different (P<0.01), between the treatment groups compared with no difference (P>0.05). (8) Quail model of hyperuricemia model have been successfully set up, model of serum uric acid was significantly higher than the blank group, the difference was significant (P<0.01). (9) Serum uric acid of Liu wei tongfeng-Drink group, Allopurinol and benzbromarone group than it’s group before treatment were significantly decreased, the difference was statistically significant (respectively P<0.05, P <0.01, P<0.01 ); drug treatment, Liu wei tongfeng-Drink group, Allopurinol group and benzbromarone group than those in model group, serum uric acid decreased, the difference was significant (respectively P<0.05, P<0.01, P<0.01); while between the treatment groups compared with no significant differences (all P>0.05). (10) Blood XOD at Liu wei tongfeng-Drink group, Allopurinol group than in their pre-medication has declined markedly, the difference was statistically significant (respectively P<0.05, P<0.01); XOD at Benzbromarone group was lower than its medication before, the fall there was no significant difference (P>0.05); After their drug treatment, blood XOD at Liu wei tongfeng-Drink group, Allopurinol group is lower than those in model group, the difference was statistically significant (P<0.05, P<0.01); blood XOD in benzbromarone group compared with the model group, that was no statistical significance (P>0.05); blood XOD at Liu wei tongfeng-Drink gout than allopurinol group, which is lower than benzbromarone group , the differences were statistically significant (P<0.05); Allopurinol group was significantly lower than blood XOD benzbromarone group, there is significant difference (P<0.01).
Conclusion: (1) Liu wei tongfeng-Drink for gouty arthritis rat model has a good anti-inflammatory and analgesic effect of swelling. (2) Model of gouty arthritis have MMP-3, IFN-γexpression in it’s joint surrounding tissue, no obvious IL-4 expression. (3)Liu wei tongfeng-Drink can reduce surrounding tissue inflammatory response by inhibiting MMP-3, IFN-γactivity of gouty arthritis model articulation surrounding tissue, which may be related to IL-4 expression has nothing to do. (4) The success of the quail hyperuricemia model was set up, their blood uric acid and xanthine oxidase increased. (5) Liu wei tongfeng-Drink can reduce serum uric acid and serum xanthine oxidase activity of hyperuricemia quail model. (6) Liu wei tongfeng-Drink can be effective prevention and treatment of gouty arthritis.
引文
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