| Objective: This experiment aimed to study anti-inflammation,antithrombosis and diuretic effect of compatibilities between Rhubarb and Fructus Aurantii Immaturus,Chinese goldthread rhizome,Moutan Cortex,peach seed and Kansui Radix stir-baked with vinegar,as well as to determine the contents of free and total anthraquinones among five different compatibilities of Rhubarb.A HPLC method was established to determine the change of serum anthraquinones.These studies on compatibility rule of herb-pair in Rhubarb reveal the specific pharmacodynamic material basis and mechanism,which provides scientific references for further study of TCM connotations.Methods: The classical method of acute toxicity was used to determine the content of five different compatibility of Rhubarb and compare their content and the acute toxicity.The effects of five different compatibility of Rhubarb on acute inflammation were observed in the mouse paw swelling induced by carrageenan.Adrenaline hydrochloride mode method was used to observe the effection of diuretic.An UV Spectrophotometry method was established to determine the contents of free and total anthraquinones among five Different Compatibility of Rhubarb and the UV spectra of the solutions extracted by three kinds of solvents(cyclohexane,chloroform and butanol)from decoction of different compounds.Determinng the five Different Compatibility of Rhubarb were carried out by HPLC:The separation was performed at 30? on a Thermo Syncronis C18 column(250mm×4.6mm,5 ?m).The mobile phase consisted of acetonitrile and 0.1% phosphoric acid aqueous solution.The flow rate was 1.0mL/min and the detection wavelength was 254 nm.The change of serum anthraquinones among five Different Compatibility of Rhubarb were determined by HPLC to observe the variation.Result: 1.The LD50 has not been determined.According to crude drug content,the MLD were 145.33,142.30,117.53,103.45,113.09,182.36 g/kg respectively,which equal to 581,569,470,418,452,729 times of people daily doses in clinic.2.The content of five different compatibility of Rhubarb with high and low dosage groups(30,15 g/kg)could inhibit the paw edema in mice.Furthermore it could reduce NO and MDA production and enhanced activity of SOD in mice inflammatory tissue.3.The five different compatibilities of Rhubarb could apparently/obviously improve the abnormality of hemorheology and blood coagulation function in acute blood stasis rats.PT and APTT were obviously reduced,and the content of TT and FIB obviously increased.Meanwhile it significantly reduced the plasma TXB2,VWF and GMP-140 level of blood stasis rats,and also had significant tendency to improve 6-keto-PGF1? level;4.After 4 hours,the urine output and the urinary levels of Na+,K+,Cl-the normal rats were significantly increased.The five different compatibility of Rhubarb could reduce the acute renal failure rats renal damage and improve its kidney function.and could signficantly decrease the levels of serum urea nitrogen,creatinine(Cr),K+,Na+ and increase urine volum of rats with ARF;5.The total and free anthraquinones were changed after compatibilities,especially when they were combined with peach seed and Kansui Radix stir-baked with vinegar.The content of variation anthraquinones was increased whereas the rest of content of anthraquinones among different compatibility of Rhubarb was all decreased.6.After IG,the five different compatibility of Rhubarb,aloe-emodin,rhein and emodin were detectable in plasma.The changes of content of serum show a certain tendency.Conclusion: The method to study acute toxicity,anti-inflammation,antithrombosis and diuretic effect in five different compatibilities of Rhubarb is safe and has low toxicity.The five different herb-pair,which have the anti-inflammation effect after compatibility,could obviously improve the abnormality of hemorheology,blood coagulation function and the urine output.The content of anthraquinones in Rhubarb was mostly decreased by different compatibilities,which may contribute to the characteristics of efficacy to Rhubarb. |
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