Study On Effect And Mechanisms Of Angelica Polysaccharide On Blood Stasis Rats Before And After Wine Broiled

Angelica sinensis is one of the most widely used drugs for promoting blood circulation and removing blood stasis,and there was a statement of“nine Angelica sinensis in each ten prescriptions”.The content of polysaccharides in Angelica sinensis is relatively high,about 10% to 15%.Angelica sinensis is traditionally prepared by the wine-broiled method.After wine-broiled,the function of passing through the channels and activating blood stasis is enhanced,it has not been reported whether the effect of Angelica sinensis after wine broiled changes due to the changes in polysaccharide composition.This experiment is to study the difference of pharmacological effects of Angelica sinensis polysaccharide before and after wine broiled on rats with acute blood stasis syndrome,trying to expiain the underlying mechanism of its therapeutic effect,establishing the research foundation of Angelica sinensis polysaccharide processed transformation theory and providing experimental evidence for clarifyingthe pharmacological effects of Angelica sinensis polysaccharide components and chosing medication for clinical blood stasis syndrome.Experimental methods:1.In accordance with the fourth part of the “Chinese pharmacopoeia“(2015 edition)general rules,add yellow rice wine to the raw Angelica decoction in a certain proportion,and put it in a wok until it is fried to become golden brown.Raw Angelica total polysaccharides and wine broiled Angelica total polysaccharides were obtained through hot water reflux extraction,alcohol precipitation,sevage method to remove protein and distilled water dialysis of the raw Angelica decoction pieces and wine broiled Angelica decoction pieces.Taking glucose as reference,using the phenol sulfuric acid colorimetric method to measure the content of polysaccharides equivalent to glucose in the total polysaccharides of raw Angelica sinensis and wine broiled Angelica sinensis,The sulfuric acid-carbazole method was used to detect the content of uronic acid in the total polysaccharides of Angelica Sinensis and wine broiled Angelica Sinensis.2.Acute cold coagulation stasis model in rats was caused by subcutaneous injection of epinephrine and ice water bath,rats were orally administered with low-dose(400 mg/kg)and high-dose(800 mg/kg)aqueous solutions of raw Angelica polysaccharides and wine broiled Angelica polysaccharides for intervention treatment.The effect of Angelica sinensis polysaccharides before and after wine broiled on hemorheological indexes in rats with acute blood stasis syndrome was measured by a hemorheology detector,the influence of four functional indexes of blood coagulation in the plasma of rats in each group was measured by automatic coagulation instrument,Hematoxylin-eosin HE staining is to observe the pathological changes of heart and lung tissue in rats,calculating the changes of thymus index and spleen index of immune organs in each group of rats and detect the changes in oxidative stress indicators which are the content of total superoxide dismutase T-SOD,glutathione peroxidase GSH-px and MDA in rat plasma at the same time.3.In vitro experimental model of oxidative damage of human umbilical vein endothelial cells induced by hydrogen peroxide.Detecting the effect of Angelica sinensis polysaccharides on the value-added activity of HUVEC induced by hydrogen peroxide by cck-8 method,detecting the changes in the content of nitric oxide NO and lactate dehydrogenase LDH in the supernatant of each group of cells,studying the anti-oxidation protective effect of Angelica sinensis polysaccharides before and after wine broiled on HUVEC at the cell level.Experimental results:1.Raw Angelica decoction pieces are light yellow and flexible,while wine broiled Angelica decoction pieces are dark yellow-brown and hard with a slight burnt spot;raw Angelica polysaccharides are yellow-white powder,while wine broiled Angelica polysaccharides are dark yellow powder.The extraction yield of raw Angelica sinensis total polysaccharides was 13.9%,the extraction yield of wine broiled Angelica sinensis total polysaccharides was 15.6%,of which the polysaccharide content in raw Angelica sinensis total polysaccharide was 83.1%,and the polysaccharide content in wine broiled Angelica sinensis total polysaccharide was 86.5%;the content of uronic acid in the total polysaccharide of raw angelica sinensis was 40.2%,while the content of uronic acid in the total polysaccharide of wine broiled angelica sinensis was 43.6%.2.Compared with the model group,both the raw Angelica polysaccharide groups and the wine broiled Angelica polysaccharide groups can reduce the whole blood viscosity(WBV),plasma viscosity(PV)and the content of fibrinogen,prolong prothrombin time(PT),activated partial thrombin time(APTT)and thrombin time(TT)in plasma of blood stasis rats(P <0.05,P <0.01);compared with the results of thepositive drug compound Danshen dripping pills group,the effect of the function of high-dose group of raw Angelica sinensis polysaccharides to improve blood rheology and coagulation system in rats are close to it,and the difference is not statistically significant(P >0.05),while the effect of low-dose and high-dose wine broiled Angelica polysaccharides groups are better than that of the compound Danshen dripping pills group(P <0.05).Both raw Angelica polysaccharides and wine broiled Angelica polysaccharides can improve myocardial cells widened and heart muscle cell nucleus rounds,alveolar wall thickening,pulmonary interstitial edema,and pulmonary hemorrhage,etc.Among them,the effect of polysaccharides from wine broiled Angelica is better than polysaccharides from raw Angelica sinensis3.The thymus index and spleen index of the model group rats were significantly increased(P <0.01).After administration,the effect of raw Angelica polysaccharide on the thymus index and spleen index of blood stasis rats was not statistically significant(P >0.05),while wine broiled Angelica polysaccharides significantly reduced the thymus index and spleen index of rats(P <0.05).4.The MDA content in the plasma of the model group rats was significant increased,and the activities of total superoxide dismutaseT-SOD and glutathione peroxidase GSH-Px were significantly reduced(P<0.01).Compared with the model group,the MDA content of the raw Angelica polysaccharide groups and the wine broiled Angelica polysaccharide groups decreased,and the activities of T-SOD and GSH-px increased significantly(P <0.05,P <0.01);compared with the results of the compound Danshen dripping pills group,the effect of improving the oxidative stress level of rats in raw Angelica polysaccharide groups were similar to that of the compound Danshen dripping pill group and and the difference was not statistically significant(P >0.05),while the effect of enhancing the activity of GSH-px in the low-dose group of wine broieed Angelica Sinensis polysaccharidethe and effect of enhancing the activity of T-SOD and GSH-px in the high-dose group of wine broiled Angelica Sinensis polysaccharide were better than that of compound Danshen dripping pills group(P <0.05).5.The raw Angelica sinensis polysaccharide had no significant effect on the proliferation activity of HUVEC damaged by hydrogen peroxide(P >0.05),while the wine broiled Angelica sinensis polysaccharide significantly enhanced the proliferation activity of injured HUVEC(P<0.05).There was no significant difference in the effect of polysaccharides from raw Angelica sinensis on the activity of NO in HUVEC damaged byhydrogen peroxide(P >0.05).while polysaccharides from wine broiled Angelica sinensis could significantly increase the content of NO released by HUVEC(P <0.05).Both raw Angelica polysaccharides and wine broiled Angelica polysaccharides could effectively reduce the LDH leakage from HUVEC cells damaged by hydrogen peroxide(P <0.05,P<0.01).Experimental conclusion:Polysaccharides in Angelica sinensis are easier to dissolve after wine broiled,the content of polysaccharide in wine broiled Angelica sinensis total polysaccharides increased.The effect of wine broiled Angelica sinensis polysaccharides that improving blood rheology,blood coagulation system function and histopathological morphology in rats with acute blood stasis are better than raw Angelica sinensis polysaccharides,the mechanism were probably related to the changes of polysaccharide content afeter wine broiled,the improving of the level of oxidative stress,immune function and vascular endothelial cell damage in rats.At the same time,it has also been proved that the polysaccharide component should be the material basis of Angelica sinensis for promoting blood circulation and removing blood stasis and its effect could be enhanced after wine broiled.Polysaccharide conversion and enhanced activity should be one of the connotative mechanisms of wine broiled of the Angelica sinensis.