Preclinical Research On Xiaofeng Powder Treatment Of Gouty Arthritis And Its Mechanism Of Cartilage Matrix Degradation

Objective In order to explore the toxicity and efficacy of Xiaofeng Powder in vivo,we take acute toxicity,long-term toxicity.Hyperuricemia,Gouty arthritis,Inflammation and pain model.On other hand,we explore the effect of gouty arthritis induced-cartilage degradation in vitro on the basis of animal experiments as clinical references.Methods Toxicoloagy:To observation of mortality rate by administered Xiaofeng Powder twice(crude drug 175.7 g·kg-1 x2)within 24 hours in mice that can calculate the mice LD50 and MTD.To observation of coaogulation,blood routine,blood biochemical parameters,organ coefficient and histopathological examination by administered Xiaofeng Powder(crude drug 200.100,50 g·kg-1)three consecutive months and withdrawal two weeks.Pharmacodynamics:To observe the effect of blood uric acid,urine uric acid,serum xanthine oxidase and liver xanthine oxidase in hyperuricemia rats and mice which are ig Xiaofeng Powder.To observe the effect of joint swelling,joint lesions,synovial fluid WBC,IL-1? and IL-6 in Gouty arthritis rabbits and rats which are ig Xiaofeng Powder.In addition,we observe the xylene-induced mice ear edema model,acetic acid-induced mice peritoneal capillary permeability model,paper-induced granuloma in rats model and acetic acid-induced writhing response model to validation of anti-inflammatory and analgesic effects for Xiaofeng Powder.Mechanism:To observe the effect of cell viability,iNOS,NO,IL-1?,TNF-? in MSU stimulated chondrocytes for Xiaofeng Powder containing serum.To observe the effect of GAGs,Hyp and proteoglycans in MSU stimulated chondrocytes for Xiaofeng Powder containing serum that can evaluate the extent of the cartilage matrix proteoglycan and coiiagen degradation.Moreover,we observe the effect of MMP-3,TIMP-1,ADAMTS-4 and TIMP-3 in MSU stimulated chondrocytes for Xiaofeng Powder containing serum that can research the mechanism of inhibition of cartilage matrix degradation.Results Toxicology:There were no mice dead or abnormal after fourteen days from administration of Xiaofeng Powder that mice LD50 can't be counted.The maximum dose in mice was crude drug 351.4 g·kg-1,equivalent to 175.7 times of clinical adult dosage.We found rats generally in good condition,appearance,behavior,excreta no abnormal changes,and rats weight,food intake,coagulation system,blood routine,blood biochemical parameters,organ coefficient and histopathological examination were no drug-induced toxicity after three consecutive months from administration of Xiaofeng Powder three groups(crude drug 50,100.200 g·kg-1).Its equivalent to twenty-five,fifty,a hundred times of the clinical adult dosage.Pharmacodynamics:Xiaofeng Powder can decrease blood uric acid,urine uric acid,24 hours urinary uric acid,serum XO in hyperuricemia model(P<0.01,P<0.05),and continuous administration Xiaofeng Powder for five days is better than a single dose about lowering blood uric acid.Xiaofeng Powder can decrease joint swelling,IL-1?,IL-6,WBC,articular lesions in Gouty arthritis model(P<0.01,P<0.05).In addition,Xiaofeng Powder can decrease xylene-induced ear edema in mice,acetic acid-induced mice peritoneal capillary permeability,granulomas induced by paper,acetic acid-induced writhing in mice and prolong the first time writhing appear(P<0.01,P<0.05).Mechanism:MSU(2000 ?imol·L-1)can decrease the cell viability of chondrocytes within 24 hours(P<0.01),and MSU(1000 ?imol·L-1)can decrease the cell viability of chondrocytes within 48 hours(P<0.01).Xiaofeng Powder containing serum can increase the cell viability of chondrocytes that stimulated by MSU within 48 hours(P<0.01,P<0.05),and decrease the content of iNOS,NO,IL-1?,TNF-? after MSU stimulated chondrocytes for 48 to 72 hours(P<0.01,P<0.05).Xiaofeng Powder containing serum can decrease the activity of MMP-3 and ADAMTS-4 that can inhibit GAGs and Hyp released(P<0.01,P<0.05).In addition,Xiaofeng Powder containing serum can increase the proteoglycans expression on stained chondrocytes by Toluidine blue(P<0.01,P<0.05).Conclusion There are no toxicity about acute toxicity test and long-term toxicity tests in mice and rats which are ig Xiaofeng Powder.Xiaofeng Powder can decrease uric acid in hyperuricemia model,and inhibition of inflammation in Gouty arthritis model that owing to the decrease of serum XO,IL-1? and IL-6.In addition,we also verify the anti-inflammatory and analgesic effects of Xiaofeng Powder by Inflammation and Pain model.Described that Xiaofeng Powder not only can be applied to both acute attack and intermittent of gouty arthritis,but also can reduce uric acid to prevent the recurrence of gouty arthritis by long-term administration.To observe the mechanism of gout joint destruction reduce in vitro based on animal Gouty arthritis experiments,we first found the cell viability of chondrocytes were declined by MSU(1000?mol·L-1)after 48 hours,and the 48 hours is the best time point that.Second,Xiaofeng Powder have anti-inflammatory effects,its ralated to the activity of iNOS,NO,IL-1?,TNF-? decreased.Third,Xiaofeng Powder can inhibition of proteoglycan and collagen degradation that owing to the decrease of MMP-3 and ADAMTS-4.Xiaofeng Powder can protect chondrocytes to prevent the outflow of GAGs and Hyp from proteoglycan and collagen.