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Study on the mechanism of promoting wound healing to carbon bleeding improvement in submucosal tissue repair state of Mongolian medicine, gastric ulcer

Author: LiuWeiZhi
Tutor: CuiJian
School: Central University for Nationalities
Course: Chinese Minority Traditional Medicine
Keywords: Carbonized Agei Gastric ulcer with hemorrhage Hemostasisand coagulation Molecular mechanism
CLC: R29
Type: PhD thesis
Year: 2013
Downloads: 22
Quote: 0
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Abstract

Peptic ulcer is a common disease worldwide, and gastric ulcer is the most common type of peptic ulcer. Generally speaking, gastric ulcer itself does not cause the patient to die, but it can associated with complications such as bleeding and perforation in severe cases, and if not treated timely, the mortality rate will be very high. In gastric ulcer, bleeding is the most common complication, and it also the most common cause of upper gastrointestinal bleeding. Gastric ulcer occurs at any age, especially in young adults, and symptoms can relieve or disappeare after medication. However, qualities of gastric ulcer healing are poor and gastric ulcer can re-emerge easily, which bring great harm to the patients both physically and mentally.Hemostasis and ulcer healing promotion are key treatment methods of gastric ulcer with hemorrhage, but stagnant blood have serious impact on the quality of ulcer healing. For this reason, how to resolve problems of hemostasis after bleeding and ulcer healing after hemostasis are choke points in the treatment of gastric ulcer. Mongolian medicine have accumulated a lot of valuable experience and drugs in the treatment of gastric ulcer haemorrhage, and Agei is one of representatives. Agei which also named Xiao-Bai-Hao is the entire plants of Arteisia frigida Willd, and it is a common drug used to reduce swelling and hemostasis in Mongolian. Arteisia frigida Willd are widely used in Inner Mongolia Autonomous Regionand, Tibet Autonomous Region and Mongolia, and it has definite effect in clinical use.In preliminary studies, we found that carbonized Agei have effect on hemostasis and invigorating the circulation of blood, and this feature, which can also smmerize by "hemostasis but no stagnat of blood", is similar to the pathogenisis of gastric ulcer accompanied with haemorrhage. In this study, we put forward the hypothesis that combining hemostasis and invigorating the circulation of blood palys key role in the treatment of gastric ulcer bleeding and promotion of quality of ulcer healing. We observed the hemostatic effects of carbonized Agei and explore the hemostatic mechanism through detecting relative index in model rats, and by observing the change of VEGF, VEGFRmRNA and bFGF in ulser tissue, we also explore the mechanism of promotion of angiogenesis and ulcer healing quality. Besides, combining with immunohistochemistry and pathological method, we observed the number of new vessels, vascular density and vascular conformation, then we want to illuminate the inner relationship between ulcer and pathogenisis of haemorrhage, and we also want to establish a unified understanding between two-ways regulation treatment and treatment of gastric ulcer accompanied with haemorrhage. Eventully, this study will provide experimental basis to clinical application for ethnodrugs in the treatment of gastric ulcer accompanied with haemorrhage. Method:1. SD rats were randomly divided into8groups, named control group, model group, Yunnanbaiyao group (Positive drug), ranitidine group, Agei crude drug group, low dose carbonized Agei group, middle dose carbonized Agei group and high dose carbonized Agei group. Model rats which have acute gastric ulcer accompanied with haemorrhage were duplicated by restraint and watered stress, and intragastric administration were given7d continuously. Solvent were administrated by intragastric for control group and model group.2. Observe and analyse the area of ulcer, ulcer index and inflammatory cell infiltration by combining electron microscopy with ordinary microscope, then clear and definite the effect of carbonized Agei on promoting healing of gastric ulcer accompanied with haemorrhage.3. The mouse bleeding time was measured by severing tails test, the clotting time was measured by capillary test, and the platelet count was measured by blood biochemical detection instrument.4. Immunohistochemical method, Ⅷ factor maker, light microscope observation, new vesscels calculation and MVD detection were used in order to observe the effect of carbonized Agei on restruction of basal mucosal tissue of ulcer.5. To investigate the mechanism of hemostasis and coagulation of carbonized Agei on model rats, t-PA, PAI-1, TXA2,6-ke to-PGFla and GMP-140were measured by ELISA, and red cell assembling index, hematocrit and whole blood viscosity were measured by automatic blood rheometer.6. To explore the molecular mechanism for carbonized Agei to promote angiogenesis and the growth of granulation tissue, the level of protein expression and mRNA expression of VEGF, VEGFR, bFGF and FGFR were measured respectively by using RT-PCR and Western blot.Result:1. Carbonized Agei can significantly reduce the gastric ulcer model rat’s ulcer area and the ulcer index (p<0.05), decrease the submucosal tissue inflammatory cells embellish. Other pathological changes compared with model group have different degree of ease2. Carbonized Agei can reduce ulcer bleeding points (P<0.05) of gastric ulcer model rat; Shorten the bleeding time and blood coagulation time in rat models (P<0.01). The effect is better than that of dried medicinal herbs group; the model rat platelet count (P<0.05) is improved.3. Carbon medicine high dose group could significantly increase the number of new blood vessels of submucosal tissue (P<0.05) and carbon medicine low dose group of model rat have significant different influence of microvascular density (P<0.05).4. Carbonized Agei high dose group could increase rats serum TXB2level (P<0.05) in gastric ulcer hemorrhage model and has no effect on6-keto-PGFla (P> 0.05) in rat serum. It can significantly improve the level of GMP-140(P<0.01) in rat serum.5. Carbonized Agei high and middle dose group could significantly improve the level of t-PA (P<0.05) in model rats serum, low and high dose group can reduce the level of PAI-1(P<0.05) in model rat serum.6. Carbonized Agei has no statistical significance on erythrocyte aggregation index, hematokrit and Whole blood viscosity (P>0.05) in the gastric ulcer rat model. Ranitidine can significantly reduce rat red blood cell volume and whole blood viscosity under different shear rate (P<0.05)7. Carbonized Agei low dose group could significantly increase VEGF and its receptor protein expression levels (P<0.05) and increase VEGFmRNA expression. It can improve the expression level of bFGF, its receptor protein and mRNA.Conclusion:1. Carbonized Agei has significant effect on wound healing on the ulcer bleeding model rats:it can significantly reduce the gastric ulcer model rat’s the ulcer index, decrease the submucosal tissue inflammatory cells embellish and promote the healing of the ulcer surface; Carbonized Agei can reduce ulcer bleeding points of gastric ulcer model rat; Shorten the bleeding time and blood coagulation time in rat models; and through the increased platelet count, the bleeding part’s hemostatic and coagulant is sped up and Carbonized Agei has a good therapeutic effect on ulcer bleeding; Carbonized Agei could significantly increase the number of new blood vessels of submucosal tissue and microvascular density; stimulates the growth of granulation tissue in the ulcer area and improve the quality of ulcer healing. Carbonized Agei could improve the quality of ulcer healing.2. Carbonized Agei has a two-way adjustment to hemostatic and invigorating the circulation of blood:On the one hand it can affect blood coagulation system and play a role in hemostasis and coagulanting; On the other hand by adjusting the fibrinolytic system it could prevent thrombus formation, at the same time it has no effect on model rats hemorheology indexes and no blood stasis formation when there is too much hemostasis.3. Carbonized Agei could significantly increase VEGF and its receptor protein expression levels and increase VEGFmRNA expression. It can improve the expression level of bFGF, its receptor protein and mRNA. It reveals one of the possible molecular mechanisms of submucosal tissue reconstruction and promoting angiogenesis.

Study on the mechanism of promoting wound healing to carbon bleeding improvement in submucosal tissue repair state of Mongolian medicine, gastric ulcer

Abstract

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