| Objective In this study,the characteristics of Xinjiang Uygur Medicine-Chamomile were chosen as the objective.We have established the method of chamomile bisabolane derivatives,total saponins and flavonoids,improved the quality control standard code in Uygur medicine;To establish a method for the determination of quercetin,luteolin and apigenin in rat plasma and pharmacokinetics in rats,and to reveal the dynamic behavior of three components in rats;Preliminary pharmacological studies on reducing blood pressure,relaxing blood vessels,lowering blood fat,anti thrombus,anti-oxidation,hypoglycemic and anti tumor were investigated;To establish a method for extraction and isolation of total flavonoids from chamomile,and to investigate its pharmacodynamics.Method1.Determination of chamomile bisabolane derivatives,total saponins and flavonoids:1)GC method was used to determine the content of the ramification of opopanax in chamomile;2)Using UV method to determine the content of total saponins in chamomile;3)Using HPLC method and determining the content of luteoloside,apiin,quercetin,luteolin and apigenin in chamomile.2.Studies on the pharmacokinetics of chamomile:1)The HPLC-UV method was established for the simultaneous determination of quercetin,luteolin and apigenin in plasma of SD rats;2)The time concentration curves of quercetin,luteolin and apigenin were obtained after oral administration of SD in rats,Kinetica4.4 software was used to calculate the pharmacokinetic parameters of 3 components in rats.3.Preliminary pharmacological studies of chamomile:1)The rabbits were injected with drugs(1.5 mg/kg)through the ear vein,all rabbits were collected blood pressure data before and after administration,analysis the antihypertensive effect;2)Observed the relaxation effect of chamomile on KCL induced and NE induced vasoconstriction,and calculate the EC50 value;3)High fat rats by gavaged for 4 weeks of drug,collected rat blood,using automatic biochemical analyzer of TC、TG、HDL-C、LDL-C levels were measured;4)The effect of Chamomile on thrombosis was observed by the experiment of rat oneway vein bypass thrombosis and mouse tail vein thrombosis test.5)The healthy Kunming mice were administrated with intragastric administration of 21d for a period of time.Serum SOD,MDA,GSH-PX values were determined by the kit method.6)The effect of Chamomile on blood glucose and glucose tolerance in mice:①The normal mice were given the medicine or distilled water treatment for 7 days,the blood glucose test paper was used to measure the blood glucose and OGTT experiment at the same time;②The model of diabetic mice was established by using four alloxan,after successful modeling 3 days,Ig was administered 1 times a day for a total of 30 days,after modeling,0,10,20,and 28 days,test the blood glucose by blood glucose test paper,and after modeling 30 days,OGTT test.③Using STZ modeLing method,after successful modeling 3 days,Ig was administered 1 times a day for a total of 30 days,after modeling,0,10,20,and 28 days,test the blood glucose by blood glucose test paper,and after modeling 30 days,OGTT test.7)Using CCK-8 method,the survival rate of Hela cells was measured under different concentrations and time.Result1.Determination of chamomile bisabolane derivatives,total saponins and flavonoids:1)The content of bisabolane derivatives in chamomile was 0.19%by GC method;The content of total saponin content in chamomile was 8.02%by UV method;The content of luteoloside and apiin was 1.19 mg/g and 23.35 mg/g by HPLC method,respectively.The content of quercetin,luteolin and apigenin was 3.38 mg/g,0.50 mg/g and 0.81 mg/g by HPLC method,respectively.2、Studies on the pharmacokinetics of chamomile:1)The linear equations for determination of quercetin,luteolin and apigenin in rats of Chamomile were Y=0.3736 X0.0309(r=0.9996);Y=0.6176 X0.0127(r=0.9980);Y=0.633 X0.0471(r=0.9957).2)The pharmacokinetic parameters showed that quercetin,luteolin and apigenin peak time(Tmax)were 0.79±0.25、0.42±0.09、0.51±0.13 h.Peak concentration(Cmax)was 0.29±0.06、3.04±0.60、0.42±0.11μg/mL.0-12 hours of AUC0-12,respectively,1.79±0.29、16.00±2.38、2.91±0.74μg h/mL.Elimination half-Life(T1/2)were 13.60±6.62、4.43±1.84、8.82±4.15 h.3.Preliminary pharmacological studies of ChamomiLe1)Response surface methodology was used to determine the content of total flavonoids,the optimum extraction process of the medicinal materials was as follows:the dosage of solvent was 12 times,the extraction time was 3 times,the extraction time was 2h,and the average content of total flavonoids was 34.792mg·g-1.The best elution procedure was as follows:the selected D101 macroporous resin,the concentration of sample was 0.8269mg/mL,the maximum sample volume of 17 BV(17 times of column volume),the sample rate is 0.081BV/min,eluted with 7 BV distilled water at a flow rate of 0.081 BV/min,the eluate was removed and eluted with 6 BV of 70%ethanol at a flow rate of 0.041 BV/min.The flavonoids content in the purified solid was 52%.2)The high dose(117.1 mg/kg)of the total flavonoids of Chamomile was selected by the acute blood pressure test.3)Chamomile couLd relaxing contraction vessels by KCL,EC50=29.73(mu g/mL),relaxing contraction vessels by NE,EC50=91.09(μg/mL).4)Through the establishment of the model of hyperlipidemia,determination of alcohol extract(971 mg/kg)and water extract(1366 mg/kg),the levels of TC,TG and LDL-C in serum of rats were decreased,and the level of HDL-C was significantly increased.5)Chamomile extract(971 mg/kg)and water extract(1366 mg/kg)could significantly prolong the clotting time of normal mice,the bleeding time of mice was significantly prolonged;The ethanol extract(971 mg/kg)and water extract(1366 mg/kg)could reduce the wet weight of thrombus in rats,and the inhibition rate was about 48.76%and 31%,respectively.6)when the concentration of ethanol extract was 1.0mg/mL,the scavenging rate of DPPH free radical was about 64.28%.When the ethanol concentration was 2.5mg/mL,the scavenging rate of hydroxyl radical was 71.89%.When the ethanol concentration was 2.5mg/mL,the scavenging rate of superoxide anion was 97.42%.②High and middle dose of totaL flavonoids of ChamomiLe could significantly increased the activity of SOD and GSH-PX in serum of mice,and decreased the content of MDA.7)The effects of Chamomile on blood glucose and glucose tolerance in mice.The high dose(200mg/kg)of ethanol extract of Chamomile could significantly decrease the content of fasting blood glucose in normal mice.The high dose(971 mg/kg)of ethanol extract of Chamomile can significantly reduce the blood glucose levels of diabetic mice induced by alloxan.High ethanol extract(971 mg/kg)can significantly reduce the blood glucose levels of diabetic mice induced by STZ.In 0～120min,Chamomile can improve the glucose tolerance of the 3 model mice.8)The extract of Chamomile has obvious inhibitory effect on proliferation of cervical cancer Hela cells,the half cell inhibition rate(IC50)for 870 g/mL.Conclusion1.Based on the research of Chamomile,the optimum technology of extraction,separation and purification of flavonoids was determined,GC method was established for determination of bisabolane derivatives,UV method for the determination of total saponins and HPLC determination of galuteolin and method for content of apigenin,these methods are accurate,simple and reliable,and can be used for the extraction and determination of the effective components of Chamomile.2.Through the study of pharmacokinetics of Chamomile,HPLC method was established for the simultaneous determination of quercetin,luteolin and apigenin,and the determination of quercetin,luteolin and apigenin in rats,the pharmacokinetic parameters of Chamomile in rats were provided.The method is rapid,accurate,simple,time-saving and labor-saving,it can be used for in vitro and in vivo detection of these 3 components,and can be used to provide the basis for clinicaL application.3.Through the research on preliminary pharmacology and found that Chamomile can reduce blood pressure,relax blood vessels,lower blood fat,anti thrombus,anti oxidation,reduce blood sugar and resist tumor,the clinical application of Chamomile has been developed,which provides an important basis for the further research of pharmacology and the development of related products. |
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