Study On Screening Active Fraction Of Ardisia Gigantifolia Against Rheumatoid Arthritis

ObjectiveTo provide reference for the material foundation research and drug developmentfor rheumatoid arthritis,the active fraction of Ardisia gigantifolia was screened.Methods1.Pre-experiment of chemical constituents,determination content of total triterpenoids and systematical Separation of chemical components.Chemical constituents in Ardisia giganti folia were pre-tested by chromogenic reaction;a UV method was established for determination content of total triterpenoids with the oleanolic acid as the control;ethanolic extract from Ardisia giganti folia were mixed with diatomite,dried at 70?,smashed,and then extracted by petroleum ether(30?60?),ethyl acetate,n-butanol,water in turn.2.Screening Active Fraction of Ardisia giganti folia against Rheumatoid Arthritis and its safety study.In order to screening active fraction of Ardisia giganti folia against rheumatoid arthritis,the petroleum fraction,ethyl acetate fraction,n-butanol fraction and water fraction were used for the treatment of the mice modle with paw edema induced by carrageenan and the mice modle with the increasing of abdominal cavity capillary permeability and torsion induced by acetic acid.MH7A cells,treated with the four extract fractions,was assessed by MTT assays to evaluate the proliferation inhibition effect.The maximum dosage of mouse lavage administration was 150 times higher than the man's nomal dose to investigat the safety of total extract fraction and four extract fractions.3.Experimental studies on anti-rheumatic arthritis of petroleum fraction.Freund's complete adjuvant(FCA)was injected subcutaneously into the right hindfoot to establishthe adjuvant arthritis model of rats(AA rats).Curative effect of petroleum fraction was estimated by systemic inflammatory response syndrome(SIRS)score,arthritis index score,joint swelling count,the swelling level of secondary side joint,contents of MDA in rat serum,content of PGE2 in inflammatory side tissues.Results1.Ardisia gigantifolia contains amino acids,polysaccharides,organic acids,tannin,isocoumarin,tri-terpenoid saponins,essential oils,etc.Using oleanolic acid as the control,a UV method was established for determination content of total triterpenoids,regression equation was A=0.0654C-0.1114(R=0.9997),the calibration curve of oleanolic acid showed good linear relationship in the range of 3.333?16.667?g · mL-1,the RSD of precision,stability in 60?120min and repeatability were 0.84%,2.62%,1.43%,the average recovery was 97.64%(RSD=1.29).2.The four extract fractions could inhibit the paw edema induced by carrageenin in mice and the increase of abdominal capillary permeability induced by acetic acid in mice,showed a certain degree of anti-inflammation action.The four extract fractions could decrease the number of torsion causing by acetic acid in mice,had a certain degree of analgesic action.The petroleum fraction had a significant effect in anti-inflammation and analgesic action.The proliferation of MH7A could be suppressed by The petroleum fraction(IC50=68.1?g · mL-1)and ethyl acetate fraction(53.6?g · mL-1).The total extract fraction and four extract fractions of Ardisia giganti folia had no apparent toxicity,no mouse died and had systemic toxicity reacts after the medicine was given within 14d,no obvious pathological changes were found in mice,there was no statistically significant difference of weight between the treated groups and the blank control group.These results showed that the petroleum fraction was the active fraction of Ardisia gigantifolia against rheumatoid arthritis and its safety met the requirements.3.Each dosage group of the petroleum fraction could decrease the scores of systemic inflammatory response syndrome and arthritis index,reduced the joint swelling count,lessen the swelling level of secondary side joint of AA rats,showed the effect of ameliorating the symptoms of inflammation disorders,including rheumatoid arthritis.Each dosage group of the petroleum fractionhad obvious inhibition on MDA in rat serum,showed a strong antioxidant activity.Each dosage group of the petroleum fraction decreased the level of PGE2 in the inflammatory side tissues,inhibited the synthesis or release of PGE2· Each dosage group of the petroleum fraction decreased the spleen index and the thymus index of AA rats,showed a certain level of the immunosuppressive function.Each dosage group of the petroleum fraction could decrease the liver index of AA rats.ConclusionThese results show that the petroleum fraction is the active fraction of Ardisia gigantifolia against rheumatoid arthritis and it has no obviously toxic side effect.The studycan provide reference for the material foundation research and drug development for rheumatoid arthritis.