Studies Of Quality Standard And Pharmacodynamics On Weitong Injection

BackgroundPost-surgical adhesion in abdominopelvic cavity means some indeterminate range of adherences or fibrous bands which are caused by surgery or inflammation between the normally separated organ and abdominal wall or organs.According to previous studies,more than 93%patients who have had undergone surgery suffer post-surgical adhesion.The main way to prevent and cure post-surgical adhesion is having a second operation and using some anti-inflammatory drugs.However,a second operation can add suffering to the patients and the curative effect of many anti-inflammatory drugs needs a further evaluation.Recently,a number of TCM (Traditional Chinese Medicine) treatment and herbal preparation have been evaluated for their value in post-surgical adhesion prevention.So it is necessary to exert the superiority and characteristic of TCM in treating post-surgical adhesion.WTI(Weitong Injection),developed by Department of pharmaceutics,Nanfang Hospital,is a pure herbal injectable powder for anti-adhesion.It bases on the study of Changtong Oral Liqud and contains components from herb,which can improve microcirculation of blood,decrease inflammatory reaction and exudation.It is another important new drug for post-surgical adhesion.ObjectiveTo detect the main components of WTI by HPLC and titration.To observe the acute toxic reaction of WTI in mouse.To study the preventive and curative effect of WTI on post-surgical abdominal adhesions in rat and rabbit. Methods1.Detection of components in WTIComponents of WTI were determined by HPLC and titration.The mobile phase of D was phosphate buffer solution:methanol(95:5).The mobile phase of Q was 1%phosphoric acid:acetonitrile(60:40).Flow rate was 1mL·min^-1.The UV detective wavelength was 220nm.Chromatographic column was Luna^? 5u C18(2) 100A.Column temperature was room temperature.Caustic soda can cause the hydrolysis of the two esterfunction on saponin which is a main component of WTI. Then use the hydrochloric acid to neutralize the excess caustic soda.2.The acute toxic reaction of WTI in mouse.Fifty mice were chose and randomized into five groups.After injected with different concentrations of WTI,the mice's action and toxic reaction could be observed.Base on the information of mice's death,the LD₅₀ of WTI could be calculated.3.Pharmacodynamic action of WTIAnimal(rats and robbits) was received laparotomy and prepared as intestinal adhesion model.WTI was injected for 7 days.Dexamethasone were used as positive control.Blood routine examination was conducted.Intraperitoneal adhesions were graded by adhesion score on day 7.Tissues from normal and adhesive tissues were collected to prepare pathological sections.Result1.The analytic research of main components in WTI.The content of D in WTI is 4.99%,Q-A is 14.98%and Q-B is 11.42%.The method of HPLC used would not be interfered by other components and it has good precision,recovery,stability.The range of linearity could satisfy the determination.By the method of titration,the content of saponin in WTI is 51.20%.2.The LD₅₀ of WTI is 8.4mg/Kg.3.Pharmacodynamic action of WTI3.1 Preventive effect of WIT on post-surgical abdominal sdhesions in rats3.1.1 The effect of WTI on adhesive degree of rats. It has significant difference among the six groups.The sequence of adhesive degree from low to high is:normal control group＜high dose WTI group＜medium dose WTI group＜positive control group＜low dose WTI group＜model control group.3.1.2 The effect of WTI on blood routine examination of rats.Except that of low dose WTI group,White Blood Cell(WBC) of model control group has significant difference from that of other groups.WBC of low dose WTI group is significant less than that of normal control group.Compared to that of low dose WTI group and normal control group,Red Blood Cell(RBC) of positive control group is significant different.RBC of normal control group and low dose WTI group have significant difference than that of model control group.There is a marked difference on RBC between normal control group and low dose WTI group.Blood platelet(PLT) of model control group has no significant difference from that of low dose WTI group,however,it has significant difference from those of other groups. Compared to that of other groups,positive control group' PLT is significant more. Lymphocyte(LYM) of high and medium dose WTI groups are significant different than that of model control group and normal control group,but significant more than that of positive control group.3.1.3 The effect of WTI on Hyp in abdominal wall tissue of rat.Compared to that of model conrol group,WTI can significantly decease Hyp in abdominal wall tissue of rat.3.1.4 The observation of rat pathological sectionWTI can decrease the number of inflammatory cell and the hyperplasy of fibrous tissue.3.2 Preventive effect of WIT on post-surgical abdominal sdhesions in robbits.3.2.1 The effect of WTI on adhesive degree of robbits.It has significant difference among the six groups.The sequence of adhesive degree from low to high is:normal control group＜high dose WTI group＜medium dose WTI group＜positive control group＜low dose WTI group＜model control group. 3.2.2 The effect of WTI on blood routine examination of rats.Compared to that of model conrol group,WTI can significantly decease WBC,RBC,LYM and FbgC.3.2.3 The effect of WTI on Hyp in abdominal wall tissue and urine of robbits.Compared to that of model conrol group,WTI can significantly decease Hyp in abdominal wall tissue and urine of robbits.3.2.4 The effect of WTI on cell factor of robbits.Compared to that of model conrol group,WTI can significantly decease t-PA and PAI-1 in blood plasm of robbits;Compared to that of model conrol group,WTI can significantly decease IL-1βand TGF-βin blood serum of robbits.Conclusions1.D and Q were the main components of WTI.2.The security of WTI is satisfactory.The LD₅₀ got from the experiment can provide more evidences to the pharmacological and pharmacodynamics research.3.WTI could reduce excess synthesis of collagen in traumatized peritoneum and decrease fibrosis level.WTI could also reduce inflammatory reaction and improve the high blood viscosity caused by trauma.