ABSTRACT
ObjectiveThe therapeutic effect of polysaccharides from Zanthoxyli Pericarpium on Alzheimer's disease（AD） was evaluated through establishing a mouse model of AD， and the structural characteristics of the polysaccharides was analyzed by sugar spectrum. MethodThe AD model of mice with rapid aging was established by intraperitoneal injection of D-galactose combined with gavage of aluminum trichloride， and the learning and memory ability of mice was evaluated by Morris water maze test， the histopathological status of brain and neuronal damage were observed by hematoxylin-eosin（HE） staining and Nissl staining. After hydrolysis of polysaccharides from Zanthoxyli Pericarpium with acid and different glycosidases， the characteristics of hydrolysates were analyzed by high performance thin layer chromatography（HPTLC） and fluorescence assisted carbohydrate gel electrophoresis（PACE）. HPTLC chromatography was performed on a silica gel 60 plate with sampling volume of 5 μL， developing solvent of ethyl acetate-glacial acetic acid-water（2∶2∶1）， developing twice， aniline-diphenylamine-phosphoric acid solution as chromogenic agent， and heating at 105 ℃ for 10 min， and then observed under sunlight. PACE experimental conditions were 34% separation gel and 8% concentration gel， electrophoresis buffer was 0.1 mol·L-1 tris（hydroxymethyl） aminomethane（Tris）-boric acid buffer（pH 8.2）. Electrophoresis was carried out at 0 ℃ and the loading amount was 3-6 μL. The sample ran to the front of the gel with a constant current of 15 mA， and imaged under ultraviolet 365 nm. ResultThe results of Morris water maze test showed that polysaccharides from Zanthoxyli Pericarpium significantly improved the learning and memory ability of AD model mice， shortened the escape latency， and significantly increased the number of crossing and the residence time in the target quadrant. The results of histopathological experiments showed that polysaccharides from Zanthoxyli Pericarpium could improve the pathological conditions and neuronal damage in the CA1 and CA3 regions of hippocampus of AD mice， and the number of Nissl corpuscles was significantly increased. The results of sugar spectrum analysis showed that the results of HPTLC and PACE analysis were basically consistent， polysaccharides from Zanthoxyli Pericarpium could be mainly hydrolyzed into small molecular sugars by cellulase and pectinase， indicating that they mainly contained β-1，4-glucosidic bond and α-1，4-galacturonic acid glycosidic bond， and could be slightly hydrolyzed by glucanase， β-galactosidase and β-mannase， indicating that they contained only a small amount of α-1，6-glucosidic bond， β-galactosidic bond， β-1，4-mannosidic bond. ConclusionPolysaccharides from Zanthoxyli Pericarpium has obvious therapeutic effect on AD mice， and its structure mainly contains β-1，4-glucosidic bond and α-1，4-galacturonic acid glycosidic bond， which can provide a reference for the structural analysis of traditional Chinese medicine polysaccharides.
ABSTRACT
OBJECTIVE：To study the difference of dissolution performance of chemical component and anticonvulsant effect of Bombyx mori decoction and powder taken with water，and to provide reference for the selection of application forms of B. mori. METHODS：The yield of dry extract was determined for decoction and powder biomimetic gastric juice of B. mori. HPLC method was used to detect the content of ammonium oxalate in decoction and powder biomimetic gastric juice of B. mori. The content of protein in decoction and powder biomimetic gastric juice of B. mori was determined with bicinchoninic acid（BCA）method. Mice was divided into normal group（1％Sodium carboxymethylcellulose solution），model group（1％Sodium carboxymethylcellulose solution），positive group （phenytoin，2 mg/kg） and B. mori decoction and powder suspensions high-dose，medium-dose and low-dose groups（0.75，1.5，3 g/kg by crude drug）according to random number table，with 20 mice in each group. After 60 min of intragastric administration，electric stimulation was conducted，and the rate of convulsion in mice was recorded. RESULTS：The yields of dry extract were 22.08％ and 26.40％ in decoction and powder gastric juice of B. mori （P＜0.05）；the contents of ammonium oxalate were 11.22％ and 16.83％ （P＜0.05），and the contents of protein was 3.39％ and 4.92％ （P＜0.01）. Compared with normal group，convulsion rate of mice was increased significantly in model group （P＜0.01）；compared with model group，convulsion rates of mice were decreased significantly in administration groups （P＜0.05 or P＜0.01），and the convulsion rate of mice in B. mori powder suspensions group was lower than decoction group. CONCLUSIONS：The dissolution performance of the chemical component from gastric juice of B. mori powder is better than that of decoction， and the anticonvulsant effect of B. mori powder is better than decoction.
ABSTRACT
OBJECTIVE:To optimize extraction technology of protein from Cryptotympana pustulata and investigate its in vitro antioxidant activity,so as to provide reference for further research of protein from C. pustulata. METHODS:Using extraction amount of protein as response value,based on single factor test,Box-Behnken response surface methodology was used to optimize the ratio of liquid to material,ultraonic time and extraction times.Validation test was conducted. Using Vitamin C(VC)as positive control, in vitro antioxidant activity of protein from C. pustulata was evaluated by using scavenging rate of 1, 1-diphenyl-2-picrylhydrazyl(DPPH)and 2,2′-azino-bis(3-ethyl benzothiazoline-6-sulfonic acid)(ABTS)free radical as index. RESULTS:The optimal extraction condition of protein from C.pustulata was as follows as the ratio of liquid to material 28:1(mL/g), ultrasonic time of 65 min,extracting for twice. In validation test,average extraction amount of protein from C. pustulata was 65.45 mg/g(RSD=1.68%,n=3),relative error of which to predicted value was 5.48%. The protein from C. pustulata showed strong scavenging effect on ABTS and DPPH free radicals. When the concentration of protein from C. slough was 0.2 mg/mL,and the scavenging rate of it to ABTS free radicals was 97%,the effect of which was similar to VC.The protein from C. pustulata showed weak scavenging ability to DPPH free radical,IC50was 0.96 mg/mL,the effect of which was not as good as VC. CONCLUSIONS:The extraction technology of protein from C. pustulata optimized by Box-Behnken response surface methodology shows high accuracy and good reliability.The protein from C.pustulata shows certain antioxidant activity in vitro.
ABSTRACT
OBJECTIVE:To establish HPLC fingerprint of stir-baked Manis pentadactyla.METHODS:HPLC method was conducted.The determination was performed on Capcell Pak Mg Ⅱ S5 C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution (gradient elution) at the flow rate of 0.8 mL/min.The detection wavelength was set at 275 nm,and column temperature was 30 ℃.The sample size was 10 μL.Using oxyphenylaminopropionic acid as reference,HPLC chromatograms of 11 batches of medicinal materials were determined.TCM Chromatogram Fingerprint Similarity Evaluation System (2004 A edition) was used for common peak identification and similarity evaluation.RESULTS:There were 23 common peaks in 11 batches of stir-baked M.pentadactyla,with similarity＞0.90.After validation,HPLC chromatograms of 11 batches of medicinal materials were in good agreement with control fingerprints.CONCLUSIONS:Established HPLC fingerprint can provide reference for the identification and quality evaluation of stir-baked M.pentadactyla.
ABSTRACT
OBJECTIVE:To establish HPLC fingerprint of stir-baked Manis pentadactyla.METHODS:HPLC method was conducted.The determination was performed on Capcell Pak Mg Ⅱ S5 C18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution (gradient elution) at the flow rate of 0.8 mL/min.The detection wavelength was set at 275 nm,and column temperature was 30 ℃.The sample size was 10 μL.Using oxyphenylaminopropionic acid as reference,HPLC chromatograms of 11 batches of medicinal materials were determined.TCM Chromatogram Fingerprint Similarity Evaluation System (2004 A edition) was used for common peak identification and similarity evaluation.RESULTS:There were 23 common peaks in 11 batches of stir-baked M.pentadactyla,with similarity＞0.90.After validation,HPLC chromatograms of 11 batches of medicinal materials were in good agreement with control fingerprints.CONCLUSIONS:Established HPLC fingerprint can provide reference for the identification and quality evaluation of stir-baked M.pentadactyla.