Study On The Effection Of Increasing The White Blood Cells And Chemical Constituents Of Caulis Spatholobi

Caulis Spatholobi is the dry stem of Spatholobus suberectus Dunn.It can nourish and promote blood,promote blood circulation to remove meridian obstruction.It is clinically used mainly for irregular menstruation,dysmenorrhea,amenorrhea,rheumatic arthralgia,paralysis and blood deficiency chlorosis.Modern pharmacological researches indicate that Caulis Spatholobi could improve the bone marrow depressions induced by radiotherapy or chemotherapy.Clinically,Caulis Spatholobi is also widely used to treat the leucopenia effectively that induced by radiotherapy or chemotherapy.Objective:The study is to investigate the pharmacodynamics effect on water extraction of Caulis Spatholobi increasing the white blood cells via leucopenia rats that induced by Cytoxan.Furthermore,use the macroporous resin technology to gather flavonoids and obtain a series of extractions,as well as sift the pharmacodynamics combined with the leucopenia rats.What’ s more,to discover the extraction which is much more effective and study the chemical constituents,provided experimental bases for the further development of Caulis Spatholobi.Methods:1.Study on the pharmacodynamics for water extraction from the Caulis Spatholobi with the leucopenia SD rats caused by Cytoxan.Establish the model by hypodermic injection of Cytoxan with 20mg/kg dosage and six days successively.Take the water extract of Caul is Spatholobi 12 days with the dosage of 24.3g original plant stems per kg by gavage to observe the effect on increasing white blood cells.Measure blood routine in 2th day,4th day,6th day,8th day,10th day,12th day,and figure out the datas in T-test.2.Study on the flavonoids garhering process of Caul is Spatholobi.With the flavonoids as the target constituents,and the adsorption amount as well as desorption rate as the evaluation index,choose the macroporous resins of AB-8,D101,HPD100,HPD 400,HPD 600.To optimize the suitable macroporous resins for isolating and purifying the total flavonoides from Caulis Spatholobi,and describe the elution characteristic.3.Study on the effect on extractions from the Caulis Spatholobi with the leucopenia SD rats caused by Cytoxan.A series of extractions are enriched and isolated from the 50%alcohol total extraction by the macroporous resins HPD 400 which is suitable for total flavonoides in Caulis Spatholobi.Study on the pharmacodynamics combined with the leucopenia rats caused by Cytoxan(the dosage is 100mg/kg,hypodermic injection).The research lasts 11 days and the blood is measured in 4th day,7th day,9th day,11th day.Finally,all the datas are figured out in T-test.4.Study on the chemical composition of the effective extraction JXT-D and n-butyl alcohol extraction.On the base of the effective extraction D,we isolate chemical compounds by silica gel column chromatography,gel chromatography and reverse phase silica gel column chromatography.And identify the structure by ’HNMR,12CNMR,MS.Results:1.Study on the pharmacodynamics for water extraction from the Caulis Spatholobi with the’ leucopenia SD rats caused by Cytoxan.On the 10th day,the amount of white blood cells in JXT-A group(the dosage is 24.3g original plant stems per kg)is 3.43X 109/L,compared with model group 2.13 X 109/L,which had markedly increased the white blood cells(p<0.01).On the 12th day,the amount of white blood cells in JXT-A group i s 7.86X 109/L,compared with model group 3.47 X 109/L,which also had obviously increased the white blood cells(p<0.01).2.Study on the flavonoids garhering process of Caulis Spatholobi.The HPD-400 macroporous resin with adsorption amount 768.5mg/5g,and desorption rate 90.50%,is the best suitable type for isolating and purifying the flavonoides in Caulis Spatholobi.According to the elution curve,80%alcohol can be eluted completely.3.Study on the effect on extractions from the Caulis Spatholobi with the lcucopenia SD rats caused by Cytoxan.On the 11th day,the amount of white blood cells in JXT-D group is 7.54×109/L,when the high-dosage is 12.15g original plant stems per kg,compared with model group 4.64×109/L,which had markedly increased the white blood cells(p<0.05).The medium-dosage group of 4.05g original plant stems per kg and low-dosage group of 1.35 g original plant stems per kg both had the increasing trend.The effect on increasing the white blood cells relied on the dosage and they present positive correlation.4.Study on the chemical composition of the effective extraction JXT-D and n-butyl alcohol extraction.Five chemical compounds were isolated from the effective extraction JXT-D and three of them were identified the structure as epicatechin,ononin and genistin.Among them,genistin was the first time to be found in the Caulis Spatholobi.Thellungianol,isolated in n-butyl alcohol extraction,was also the first time to be found in the Caulis Spatholobi.Conclusion:1.The water extraction of Caulis Spatholobi has markedly increased the SD rats white blood cells which are leucopenia induced by Cytoxan(P<0.01).2.The effective extraction JXT-D has markedly increased the SD rats white blood cells which are leucopenia induced by Cytoxan(P<0.05).Additionally,JXT-D is enriched by HPD-400 macroporous resin from 50%alcohol extraction of Caulis Spatholobi eluted with water,250%alcohol and 70%alcohol in turn,and collected the 70%alcohol eluate.3.The total flavonoids just as the epicatechin,may be one of the active constituents.The study of the pharmacodynamics in increasing the white blood cell and the active constituents of Caulis Satholohi show that it would have a developmental perspective in the future and should be studied further.