| Based on of development of gourt progress,animal models were built to imitate there stage of disease course including hyperuricemic without symptom,acute gouty arthritis and gout with high uric acid level,which were used to evaluate pharmacodynamic effects of Cangbai Qutong capsule.According to research on effects of CangBai Qutong Capsule and its decomposition on analgesia,anti-inflammatory and anti-trioxypurine,compatible regularity of assistant and guide herbal medicines was validated.After main drugs affirmed,fingerprint of Cangbaiqutong Capsules and a wavelength switching method for simultaneously determining content of 6 components by HPLC were established,which can evaluate drug's quality in the round.Part?:To study what kind of role uric acid play on the relationship between oxidative Stress and inflammation in peripheral and cerebral system of oxonate-induced hyperuricemic rats.Potassium oxonate was used to establish hyperuricemic model for four weeks.In 2nd and 4th week,total superoxide dismutase?T-SOD?,Gu,Zn-SOD activity and interleukin-1beta?IL-1??concentration in serum were determined respectively.In 4th week,one hour after last PO treatment,five rats of every group were given Evans Blue to test blood–brain barrier?BBB?permeability.Other brains were obtained to analysis T-SOD,Gu,Zn-SOD activity and IL-1?concentration in cerebral system.Meanwhile,brain and kidney were stained with hematoxylin and eosin?H&E?to observe pathological change.In 2nd week,both of T-SOD and Gu,Zn-SOD activity in serum increased obviously?P<0.05?in hyperuricemia rats.However,IL-1?content didn't change remarkably.In the 4th week,T-SOD activity in model group had become similar with control group,and at the same time IL-1?content in serum increased significantly?P<0.05?.Pathological section showed the structural and functional unit of the kidney had been damaged.On the contrary,both of T-SOD and Gu,Zn-SOD activity in brain increased obviously?P<0.05?,but IL-1?concentration was no significant difference between two groups.In addition,the results of Evans Blue and H&E suggested the integrity of BBB and structure of brain were not changed after PO treatment.Part?:To observe the Cangbai Qutong capsule's effects of anti–trioxypurine and liver/kidney protection.In order to prepare hyperuricemia animal models,oxonate was used by intraperitoneal injection?i.p.?.All animals were divided into different groups,which were given water,allopurinol and Cangbai Qutong capsule respectively.Until end of drug administration,uric acid?UA?level in serum and Xanthine oxidase?XOD?activity in liver/kidney were determined.Meanwhile,liver and kidney were stained with Hematoxylin and Eosin?H&E?to observe pathological changes.In both of two hyperuricemia models,Cangbai Qutong capsule could reduce UA lever in blood significantly?P<0.01?,and inhibit XOD activity in liver?P<0.05,P<0.01?.In addition,H&E showed Cangbai Qutong capsule had a catabatic effect on liver inflammatory injury and structural and functional unit of the kidney injury induced by hyperuricemia.Part?:To observe the analgesic and anti-inflammatory effects of Cangbai Qutong capsule on gouty arthritis?GA?.Acetic acid-induced writhing test,mice ear edema and formalin induced pain test were used to evaluate the analgesic and anti-inflammatory effects of Cangbai Qutong capsule.Coderre model were prepared to analyse Cangbai Qutong capsule.'s influence on IL-1?and matrix metalloproteinases?MMPs?in joint tissue of rats.Cangbai Qutong capsule could reduce the times of writhing in mice induced by acetic acid significantly?P<0.05?,inhibite the xylene-induced ear edema in mice?P<0.01?,and ameliorate lameness induced by formalin?P<0.01?.Meanwhile,Cangbai Qutong capsule made IL-1?content?P<0.01?and MMPs activity?P<0.05?decrease obviously in liver,kidney and joint of Coderre rat model.Part?:To observe Cangbai Qutong capsule's pharmacological action on acute gouty arthritis with hyperuricemia rat model.In order to prepare hyperuricemia animal models,oxonate was used by intraperitoneal injection?i.p.?.Both of oxonate and sodium urate were given to prepare acute gouty arthritis with hyperuricemia rat model.All animals were divided into different groups,which were given water,allopurinol positive control drug and Cangbai Qutong capsule respectively.Until end of drug administration,UA level in serum,XOD activity in liver/kidney and IL-1?and MMPs in joint tissue were determined.Cangbai Qutong capsule could reduce UA lever in blood significantly?P<0.01?,and inhibit XOD activity in liver and kidney?P<0.05,P<0.01?.Meanwhile,contents of IL-1?in liver/kidney and IL-1?/MMPs in joint tissue decreased in Cangbai Qutong capsule groups.Part?:To study effects of CangBai Qutong Capsule and its decomposition on analgesia,anti-inflammatory and anti-trioxypurine in order to understand compatible regularity of assistant and guide herbal medicines.Hot-plate test of mice and abdominal constriction test of mice were used to reseach effects of CangBai Qutong Capsule and its decomposition on analgesia.Xylene-induced ear edema of mice and carrageenan-induced paw edema of rats were used to reseach effects of CangBai Qutong Capsule and its decomposition on anti-inflammatory.Hyperuricemia mice induced by potassium oxazinate were used to reseach effects of CangBai Qutong Capsule and its decomposition on anti-trioxypurine.Sovereign drugs and minister drugs could show analgesia effects by oneself,while assistant drugs and envoy drugs didn't own analgesia effects alone.Compatible regularity of decomposition including sovereign+assistant drugs and sovereign+assistant+envoy drugs,and both of sovereign+minister drugs contained decomposition had analgesia effects obviously.Prescription dose of sovereign drugs had anti-inflammatory effects.However,minister,assistant and envoy drugs didn't display anti-inflammatory effects.Compatible regularity of decomposition including sovereign drugs had anti-inflammatory effects.Prescription dose of sovereign,minister,assistant and envoy drugs didn't own anti-trioxypurine effects by oneself.Once enhance dose twice,sovereign drugs showed anti-trioxypurine effects.Compatible regularity of decomposition including assistant drugs 2 and of sovereign+assistant drugs contained decomposition had anti-trioxypurine effects signifigantly.So CangBai Qutong Capsule had analgesia,anti-inflammatory and anti-trioxypurine effects.Part?:To establish fingerprint of Cangbaiqutong Capsules and develop a wavelength switching method for simultaneously determining content of 6 components by HPLC.The quantitative analysis was carried out on a column of Waters XSELECT CSH--C18?4.6 mm×150 mm,5?m?by simultaneous HPLC wavelength switching method,using a mobile phase of acetonitrile?A?-0.1%phosphoric acid?B?by a gradient eluted program with a flow rate of 1 mL·min-1at 30?.Ten batches of Cangbaiqutong Capsules were tested and analyzed by similarity comparison.The detection wavelength was set at284 nm?07 min,phellodendrine chloride??274 nm?710min,gentiopicroside??230nm?1014min,paeoniflorin??274 nm?1435 min,tetrandrine?berberine hydrochloride?paeonol?,which were used to built simultaneous determination method.Fringerprint spectrum had 20 common peaks.Cortex phellodendri,gentiana,radix paeoniae rubra and radix stephaniae tetrandraes'characteristic spectrums had been found,whose HPLC fingerprint similarity was more than 0.9 among different batches.The linear ranges of phellodendrine chloride,gentiopicroside,paeoniflorin,tetrandrine,berberine hydrochloride and paeonol fell within the ranges of 0.1501.504?g?0.7687.680?g?1.09610.960?g?0.2202.200?g?0.2962.956?g?0.03450.345?g,respectively.The recoveries?n=6?of phellodendrine chloride,gentiopicroside,paeoniflorin,tetrandrine,berberine hydrochloride and paeonol were 98.3%?99.2%?98.8%?98.8%?99.1%and 98.2%,respectively,and the relative standard deviations were1.32%?1.46%?1.08%?1.31%?1.26%?1.21%,respectively.Conclusion:The permeability of BBB and form of UA would be potential factors to decide what kind role UA play on keeping balance between anti-oxidative stress and induction of inflammatory response.Cangbai Qutong capsule had benefical anti–trioxypurine,liver/kidney protection,and analgesic and anti-inflammatory effects which was worth clinical application.Compatible regularity of minister,assistant and enovy herbal medicines could make low dose of sovereign drugs,which strengthened synergistic action on chlinical therapy effects.This method of fingerprint can evaluate drug's quality in the round,and simultaneous determination way is simple,accurate,reproducible and convenient for the quality control of Cangbaiqutong Capsules. |
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