Preclinical Pharmacodynamic Experiment Of DH001

Objective:Hemorrhoids is the most common disease in coloproctological department, thecommon clinical manifestation is bleeding, pain, infection, itching and so on. Hemorrhoidsaccounts for87.5%of all rectal diseases. As the study of the etiology and pathology ofhemorrhoids progressed, conservative treatment has been the overwhelming majority ofpatients to choice in its early stage.Otherwise, as one of the traditional Chinese medicine, it is very effective inactivating blood circulation to dissipate blood stasis, reliefing of swelling and pain, andhemostasis.By now, It is usually used for the treatment of traumatic injury, bleeding andabdominal pain, etc. In order to better explore the pharmacodynamics effect ofDragon`s blood as an external used drug, and to create a new field in the treatment ofhemorrhoids, we get started with preclinical pharmacodynamic experiment includingdetumescence, ulcer healing and antimicrobial,analgesic and hemostasis.Methods and Results:1.Detumescence: According to simulate the inflammatory reaction of hemorrhoids, weset up the animal model of anal swelling in rats with croton oil. The experiment is dividedinto six groups: negative control, model control, positive control(Ma yinglong), DH001Ⅰ,DH001Ⅱ, DH001Ⅲ. Setting up rat model with croton oil and rectal administration are atthe same time. Then calculate the anorectum swelling degrees6hours afterrectal administration. The results show that the swelling degrees of model control group issignificantly higher than that of negative control（P<0.05）, It turns out that the model issuccessful. The degrees of Ma yinglong group and DH001Ⅱ, DH001Ⅲare significantlylower than that of model control group（P<0.05）. In order to determine the permeability ofthe vessel, we first determine the dissolved quantity of Evans Blue in formamide, andweight the rectum cut off. The permeability of the vessel is equal to the content of EvansBlue in one gram of rectum. The results show that the permeability of Ma yinglong and DH001Ⅰ,DH001Ⅱ, DH001Ⅲ are much lower than that of the model group（P<0.05）.And compared with Ma yinglong,the results of DH001Ⅱand DH001Ⅲgroups in the experiments permeability of the vessel are remarkable(P<0.05).2.Ulcer healing and Antimicrobial: In order to simulate the natural and specificinfection, we set up the rabbit model of chemical and bacterial ulcer in the anus. Theexperiment is divided into five groups: blank group, positive control(Ma yinglong), DH001Ⅰ, DH001Ⅱ, DH001Ⅲ.Use acetic acid to set up the model of chemical ulcer. Measure theulcer’s maximum diameter with vernier caliper,then calculate the area(mm2). The result isthat compared with blank group,the data of DH001Ⅲ on the first day afteradministration are significantly reduced（P<0.05）.Also, Ma yinglong,DH001Ⅱ andDH001Ⅲ can significantly reduce the healing tim（eP<0.05）. On the other hand, we set upthe rabbit model of bacterial ulcer in the anus with a mixture of escherichia coli andstaphylococcus aureus. Test the pathological grade after continuously using DH001over5days. The results show that compared with blank group, abscessus in Ma yinglong,DH001Ⅱ and DH001Ⅲ are almost disappeared, the rectal mucous membrane returned to normal,only a few still have a tiny clump of mononuclear cells and neutrophils（P<0.05）.Andcompared with Ma yinglong,the results of DH001Ⅲ in the experiments above areremarkable（P<0.05）.3.Analgesic: The experiment is divided into five groups: blank group, positivecontrol(Ma yinglong), DH001Ⅰ, DH001Ⅱ, DH001Ⅲ. Use writhing law with acetic acid,then determine the time period between beginning and first writhing, also calculate thenumber of mice writhing in15minutes. The results show that the time period of all theother groups are not remarkable compared with blank group（P>0.05）.But they areremarkable in reducing the number of mice writhing（P<0.05）. On the other hand, we usehot plate method to determine the thermal resistance of mice. The results show thatcompared with blank group, DH001Ⅱand DH001Ⅲ can significantly increase the thermalresistance of mice（P<0.05）. But compared with Ma yinglong,the results of DH001ⅡandDH001Ⅲ in the experiments above are not remarkable(P>0.05).4.Hemostasis: The experiment is divided into five groups: blank group, positivecontrol(Ma yinglong), DH001Ⅰ, DH001Ⅱ, DH001Ⅲ. Use tail cutting method to determine the bleeding time and blood clotting time, and determine the number of bloodplatelet count with Animal Hematology Analyzer. After continuously using DH001over7days, DH001Ⅱ and DH001Ⅲ can significantly reduce the bleeding time andblood clotting time compared with blank group（P<0.05）, and the effect of DH001Ⅲ ismore remarkable than Ma yinglong（P<0.05）.On the other side, DH001Ⅱ and DH001Ⅲcan significantly increase the number of blood platelet count（P<0.05），this effect ofDH001Ⅱ and Ⅲ is also more remarkable than Ma yinglong（P<0.05）.Conclusion：1.Detumescence: DH001can significantly relieve the inflammatory edema,and alsocan apparently improve the permeability of the vessel in the rat’s rectum and anus; theeffect of DH001Ⅱand Ⅲ in the experiment of vessel’s permeability is more remarkablethan Ma yinglong(P<0.05);2.Ulcer healing and Antimicrobial: DH001can obviously reduce the area of theacetic acid-induced chemical ulcer, and speed up the ulcer healing; Also, DH001caninhibit the growth of Escherichia coli and Staphylococcus aureus; the effect of DH001Ⅲis more remarkable than Ma yinglong(P<0.05);3.Analgesic: DH001can significantly inhibit the pain caused by acetic acid, and canreduce the number of mice writhing; On the other side,DH001can increase the thermalresistance of mice, and can increase the pain threshold;4.Hemostasis: After continuously using DH001over few days, DH001can stopthe bleeding of severed tails of mice, and can increase the number of blood plateletcount,it hint that DH001can promote the hemostatic effect. And the effect of DH001Ⅲ ismore remarkable than Ma yinglong(P<0.05).