| Bletilla striata polysaccharide is the essential ingredient of Bletilla Striata (Thunb.) Reichb.f., which is a high polymer compound. It is composed of glucose and mannose with the ratio of 1:4. Bletilla striata polysaccharide could be an adhesive material to mucosa with a good histocompatibility, as a great membrana protectiva to gastric mucosa, restoring the gastric ulcers and protecting them from harmful factors.The study is mainly test on four topics:Part I: Preparation of Bletilla striata polysaccharideWith water as solvent extraction and absolute alcohol as ethanol precipitation and extraction is purified by organic solvents and then to be dialyzed, we can get a polysaccharide with a good quality and the yield is 13.8%. The physical and chemical identification and IR detection showed that Bletilla striata polysaccharide is composed of glucose and mannose, which is linked by the glycosidic link.Part II: Quality studies of Bletilla striata polysaccharide1. By the carzapol-sulfuric acid method, we detected wether the uronic acid was a component of the Bletilla striata polysaccharide and determined the polysaccharide's pH value. The carzapol-sulfuric acid method confirmed that there is no uronic acid component of the Bletilla striata polysaccharide. The pH determination showed that the higher the purity of polysaccharide, the closer the pH value to neutral status, and the pH value of the purified one is 7.3.2. Developed the determination of the polysaccharide by UV, with the anthrone-sulfuric acid method and dextrin T-20 as the standard reference substance, the concentration is linear with the UV absorbance very well between 15 mg·L-1-50 mg·L-1, and the linear regression equation is y=48.383x+1.6295,(r=0.9992,n=6). Investigating the color affected factors, we identified the best determination conditions: the volume ratio of anthrone-reagents and Bletilla striata polysaccharide or dextrin T-20 solution is 2:1, reaction time is 10 min, hydrolysis temperature is 100℃. In this condition, the average content of Bletilla striata polysaccharide is 63.70%.3. Established the method to determin the monosaccharides of Bletilla striata polysaccharide, with glucose and mannose as the standard reference substances, pre-column derivatization, and borax buffer of 50mmol/L-1 (pH = 10.5), the result showed that the relative ratio of glucose and mannose is 1:4.6.4. By the FASCO automatic blood flow measurement device, with three different degrees of the Shear rate, we detected the viscosity of Bletilla striata polysaccharide.The result showed that Bletilla striata polysaccharide is a typical pseudoplastic fluid, which was shear thinning. The viscosity of Bletilla striata polysaccharide decreased with the increasing of the shear rate and temperature, the RSD increased with the increasing of shear rate.Part III: Bletilla striata polysaccharide adhesion in vitro evaluation1. Built the animal model soccsessfully by absolute alcohol on KM rat.2. With the self-made separation detector, developed a method for the determination of separation, The best detective conditions was: tissues wetting time for 2 h, pressure plus on the tissues is 100 g, time of pressing was 15 min, water flow rate was 10 ml/min.The result showed that the separation of Bletilla striata polysaccharide on normal tissues and the model tissues were 50.79 g/cm2, 50.72 g/cm2, and between the two there was no significant difference (P=0.92), which was significant different from the separation of Carbopol (normal 29.96 g/cm2, Model 29.52 g/cm2) (P<0.01),which showed that the adhesion of Bletilla striata polysaccharide is superior to that of Carbopol.3. With the self-made detector for the adhesion of Bletilla striata polysaccharide to the nomal tissues and model tissues, screened the the best detective conditions: tissues wetting time is 2 h, hydrochloric acid washing rate is 20 ml/min. Anthrone-sulfuric acid, dextron T-20 as the standard reference substance, established the determination of polysaccharide of the washing water, the concentration is linear with the UV absorbance very well between 15 mg·L-1-50 mg·L-1, and the linear regression equation is: y= 0.0199x-0.0052 (r=0.9990,n=6). The result showed that the remains weight percentage of Bletilla striata polysaccharide on the normal tissues and model tissues were 60.64%, 61.16%. Between the two there was no significant difference (P=0.68).4. With the best detective conditions, investigated the affected factors of defferent pH value of wetting media, determined the adhesive of Bletilla striata polysaccharide to normal tissues and model tissues with the pH value of wetting media of 6.8 and 1.2. The result showed that when the pH of the wetting medium is 6.8,the separation of Bletilla striata polysaccharide on normal tissue and model tissues were 52.13 g/cm2, 53.12 g/cm2, there was no significant difference(P=0.40). The weight percentage of Bletilla striata polysaccharide remained on normal tissues and the model tissues were 62.36%, 63.05%, there was no significant difference (P=0.52). When the pH of the wetting medium is 1.2, the separation of Bletilla striata polysaccharide on normal tissue and model tissues were 44.57 g/cm2, 46.64g/cm2, there was no significant difference between the two (P=0.039); The weight percentage of Bletilla striata polysaccharide remained on normal tissues and the model tissues were 57.76%, 60.43%, there was significant difference between the two (P=0.046). The result showed that the adhesion of Bletilla striata polysaccharide is defferent when they are wetted by the wetting media of defferent pH value, the lower of pH value was, the lower separation and weight percentage remained on normal tissues and model tissues.Part IV: Bletilla striata polysaccharide selective adhesion in vivo evaluationDeveloped the method of barium sulfate reagent in two degrees concentration polysaccharide treated on SD rats, we detected at the time of 1.5, 3, 6, 12, 24 h by X-ray, contrasted the adhesion of the two groups of Bletilla striata polysaccharide on the normal group and the model group, and with two blank groups each. The results showed that the adhesion of Bletilla striata polysaccharide is very well both in model rats and normal rats, the adhesion of the polysaccharide in model group is much stronger (>24 h) than that in normal one (<24 h).And higher the concentration is, the stronger the ashesion is.It has not been reported that Bletilla striata polysaccharide is as a new type of membrane adhesion to tissues. The theme studied firstly on this issue, which is primarily evaluated and investigated the adhesion of Bletilla striata polysaccharide in vitro and in vivo.The study is confirmed the great adhesion of Bletilla striata polysaccharide to gastric mucosa. The result showed that the remained time in model rats is much longer than that in the normal rats, and the adhesion of Bletilla striata polysaccharide is positively correlated with the concentration, so as to provide a theoretical basis to the further research on exploring Bletilla striata polysaccharide as a new type of adhesive material.
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