| Objective: Gout is a chronic inflammatory rheumatic disease,owing to deposite of monosodium urate crystals(MSU)in joints and their surrounding organization,which is characterized with hyperuricemia,arthritis repeatedly,tophus sedimentation.Hyperuricemia is considered be main origin be of gout.When the concentration of blood uric acid in continue to rise more than its saturation monosodium urate crystal will be formed.Gout is gradually developed because of monosodium urate crystal.Gout would cause joint deformity if not treated timely and appropriately,therefore the treatment to gout is very important.At present the main drugs of uric acid-reducing included inhibitor of uric acid generate(such as allopurinol,febuxostat)and promoter of uric acid excretion(such as benzbromarone,probenecid).These drugs are failed to meet expectations and may lead to some side effects,even life threatening,although can decreased blood uric acid.Gout was attributed to the category of “arthralgia” in traditional Chinese medicine(TCM)theory in a long history.With the progress of science and technology,the treatment of gout with Chinese medicine has maked remarkable advancement.In this paper we would screened the TCM which have the role of uric acid reduction in a reliable hyperuricemia mice model,explored the optimal dose combination of TCM compound using the orthogonal design,and discussed preliminarily on its mechanism,expecting to provide more practice basis for clinic.Methods:1 The establishment of the acute hyperuricemia mice modelAfter divided randomly into three groups,Kunming mice were respectively given oxonic acid potassium salt by Intraperitoneal injection,hypoxanthine by intraperitoneal injection and oxonic acid potassium salt by subcutaneous injection,physiological saline by intraperitoneal injection.Their serum uric acid levels were detected using enzyme colorimetry.2 Screening the single TCM which have the role of uric acid-reducing2.1 Kunming mice were randomly divided into normal control group,model group,allopurinol group,astragalus group,coptis chinensis group,semen coicis group,radix paeoniae alba group,sinomenium acutum group(each group of eight).TCM group and allopurinol group were given corresponding drug gavage for 7 days.Model group and normal control group were given equal volume of normal saline.After the last administration,TCM group and allopurinol group,model group were given 500 mg/Kg hypoxanthine by intraperitoneal injection and 400mg/Kg oxonic acid potassium salt by subcutaneous injection to make model,then collected the serum to detect serum uric acid levels by enzyme colorimetry.2.2 Kunming mice were randomly divided into normal control group,model group,allopurinol group,forsythia group,honeysuckle group,scutellaria group,cremastra appendiculata group,capillary artemisia group,rhizoma alismatis group,rhizoma atractylodis group(each group of seven).TCM group and allopurinol group were given corresponding drug gavage for 7 days.Model group and normal control group were given equal volume of normal saline.After the last administration,traditional Chinese medicine group and allopurinol group,model group were given 500 mg/Kg hypoxanthine by intraperitoneal injection and 400mg/Kg oxonic acid potassium salt by subcutaneous injection to make model,then collected the serum to detect serum uric acid levels by enzyme colorimetry.3 Screening and verifying the optimal dose combination of TCM compound3.1 Screening the optimal dose combination of TCM compoundKunming mice were randomly divided into nine groups according to the orthogonal design,each group of seven,at the same time set up the model group and normal control group to test the model.TCM group were given corresponding drug gavage for 7 days.Model group and normal control group were given saline of equal volume.After the last administration,TCM group and model group were given 500mg/Kg hypoxanthine by intraperitoneal injection and 400mg/Kg oxonic acid potassium salt by subcutaneous injection to make model,then collected the serum to detect serum uric acid levels by enzyme colorimetry.3.2 Verifying the optimal dose combination of TCM compoundKunming mice were randomly divided into the TCM compound high dose group,the optimal dose combination of TCM compound group,the TCM compound low dose group,model group and normal control group(each group of seven).TCM group were given corresponding drug gavage for 7 days.Model group and normal control group were given equal volume of normal saline.After the last administration,TCM groups and model group were given 500mg/Kg hypoxanthine by intraperitoneal injection and 400mg/Kg oxonic acid potassium salt by subcutaneous injection to make model,then collected the serum to detect serum uric acid levels by enzyme colorimetry.4 Researching the mechanism of the optimal dose combination of TCM compoundKunming mice were randomly divided into the TCM compound optimal dose group,normal control group,model group and allopurinol group(each group of seven).TCM compound optimal dose group and allopurinol group were given corresponding drug gavage for 7 days.Model group and normal control group were given equal volume of normal saline.After the last administration,TCM compound optimal dose group,allopurinol group and model group were given 500 mg/Kg hypoxanthine by intraperitoneal injection and 400mg/Kg oxonic acid potassium salt by subcutaneous injection to make model,then collected the serum to detect serum uric acid levels by enzyme colorimetry,and to detect serum xanthine oxidase(XOD),adenosine deaminase(ADA)activity by Colorimetry.Anatomy of the liver in mice,using colorimetry to detect liver xanthine oxidase(XOD),adenosine deaminase(ADA)activity.Results:1 The establishment of the acute hyperuricemia mice modelThe level of uric acid in normal control group,hypoxanthine combine oxonic acid potassium salt group,oxonic acid potassium salt group was statistically difference(P<0.05).The uric acid level in hypoxanthine combine oxonic acid potassium salt group was significantly higher than that of normal control group,the difference was statistically significant(P<0.05).The level of uric acid in oxonic acid potassium salt group and normal control group was no significant difference,no statistical significance(P>0.05).2 Screening the single TCM which have the role of uric acid reductionThe level of serum uric acid in model group was obviously higher than that of normal control group,the difference was statistically significant(P<0.05).The level of serum uric acid in semen coicisis group,forsythia group,rhizoma atractylodis group,and allopurinol group was significantly lower than the model group,the difference was statistically significant(P<0.05).The level of serum uric acid in astragalus group,coptis chinensis group,radix paeoniae alba group and sinomenium acutum group compared with model group was no difference and no statistical significance(P>0.05).Honeysuckle group,scutellaria group,cremastra appendiculata group,capillary artemisia group and rhizoma alismatis group compared with model group,the uric acid level had no obvious change,there was no statistically significantdifference(P>0.05).The semen coicisis group compared with the astragalus group,coptis chinensis group,radix paeoniae alba group and sinomenium acutum group,the uric acid level declined obviously,the difference was statistically significant(P<0.05).The forsythia group compared with honeysuckle group,scutellaria group,cremastra appendiculata group,capillary artemisia group and rhizoma alismatis group,the level of serum uric acid declined obviously,the difference was statistically significant(P<0.05).The rhizoma atractylodis group compared with honeysuckle group,scutellaria group,cremastra appendiculata group,capillary artemisia group and rhizoma alismatis group,the level of serum uric acid declined obviously,the difference was statistically significant(P<0.05).The uric acid level in semen coicisis group,forsythia group,and rhizoma atractylodis group was higher than allopurinol group,the difference had statistical significance(P<0.05).3 Screening and verifying the optimal dose combination of TCM compound3.1 Screening the optimal dose combination of TCM compoundScreening the optimal dose combination of three kinds of TCM by the orthogonal design : Semen coicis group(4g/Kg),Forsythia group(4g/Kg),Rhizoma atractylodis group(2.5g/Kg),best of lowering uric acid.3.2 Verifying the optimal dose combination of TCM compoundThe level of serum uric acid in TCM compound high dose group,the optimal dose combination group,and low dose group was lower than the model group,the difference had statistical significance(P<0.05).Between the TCM compound high dose group,the optimal dose combination group and low dose group,the uric acid level had obvious difference(P<0.05).The uric acid level for the optimal dose combination of TCM compound dose group significantly declined,compared with the TCM compound high and low dose group was statistically difference(P<0.05),compared with the uric acid level in the orthogonal design phase decreased obviously,explained that using this dose combination was stable.4 Researching the mechanism of the optimal dose combination of TCM compoundThe uric acid level in model group was obviously higher than that of normal control group(P<0.05).Compared with model group,the level of serum uric acid in the optimal dose combination of TCM compound group and allopurinol group declined(P<0.05).The liver XOD,ADA activity and serum XOD,ADA activity in model group was significantly higher than normal control group(P<0.05).The liver XOD,ADA activity and serum XOD,ADA activity in the optimal dose combination of TCM compound group and allopurinol group obviously declined,compared with model group had difference(P<0.05).The optimal dose combination of TCM compound group was higher than allopurinol group,but no difference(P>0.05).Conclusions:1 A stable mice model of acute hyperuricemia can be established by intraperitoneal injection of hypoxanthine 500mg/Kg and subcutaneous injection of oxonic acid potassium salt 400mg/Kg.2 The optimal dose combination of three TCM by the orthogonal design was screeninged,that is Semen coicis(4g/Kg),Forsythia(4g/Kg),Rhizoma atractylodis(2.5g/Kg).3 The compound recipe of optimal dose combination of TCM can obviously reduced the blood uric acid level in acute hyperuricemia mice,and the possible mechanism is to inhibit the activity of XOD,ADA in liver and blood. |
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