CC BY
811. Date-Ito A., Kasahara K., Sawai H.,etal.(2002) Rapid evolution of the male-specific antibacterial protein andropin gene in Drosophila. J. Mol. Evol.54:665-670
12. Suttmann H, Retz M, Paulsen F, Harder J, Zwergel U, et al. (2008)Antimicrobial peptides of the Cecropin-family show potent antitumor activityagainst bladder cancer cells. BMC Urol 8: 5.
13.McKenna MP, Hekmat-Scafe DS, Gaines P, et.al. (1994)Putative Drosophilapheromone-binding proteins expressed in a subregion of the olfactory system.J. Biol. Chem.
14.Galindo K., Smith D.P..(2001) A large family of divergent Drosophila odorant-binding proteins expressed in gustatory and olfactory sensilla.Genetics, 159:1059-1072
15.Lou Y G, Cheng J A. Chemical sensory mechanism of insects. ChineseJournal of Ecology, 2001, 20(2): 66-69. (in Chinese)
16.Li S, Picimbon J F, Ji S, Kan Y C, Qiao C L, Zhou J J, Pelosi P.Multiple functions of an odorant-binding protein in the mosquitoAedes aegypti. Biochemical and Biophysical ResearchCommunications, 2008, 372(3): 464-468.
17.Damberger F F, Ishida Y, Leal W S, Wuthrich K. Structural basis ofligand binding and release in insect pheromone-binding proteins:NMR structure of antheraea polyphemus PBP1 at pH 4. 5. Journal ofMolecular Biology, 2007, 373(4): 811-819.
18.Damberger F F, Ishida Y, Leal W S, Wuthrich K. Structural basis ofligand binding and release in insect pheromone-binding proteins:NMR structure of antheraea polyphemus PBP1 at pH 4. 5. Journal ofMolecular Biology, 2007, 373(4): 811-819.
19. ZHOU Xian Ju.The effect of olfactory Behavior and olfactory input blockade and auditory defect oncircadian rhythm of locomotor activity in Drosophila [D]. PhD thesis, Shanghai Institutes for Biological Sciences (SIBS) of Chinese Academy of Sciences (CAS).2005
20.Fengliang Jin, Xiaolin Dong, Cloning and Prokaryotic Expression of cDNA Encoding General Odorant Binding Protein I from Spodoptera litura [J], SCIENTIA AGRICULTURA SINICA, 2009,42 (3).
The research of the treatment role of anemarrhena water decoction on the rat with chronic emotional stress by the empty bottles
Yunfeng DOU, Bing ZHANG, Tingli LI HeilongjiangUniversity of Chinese Medicine, Harbin,150040,China
Abstracts: Objective :To research the treatment role of Anemarrhena water decoction on the rats with chronic emotional stress induced by empety bottles.Methods:Making use of empty bottles stimulatecausethe chronic emotional stress model,Then observed behavioural changes(attacking,exploring,grooming) and weighted the rats by electronic animal scales befor the experiment,in the seventh day in the stimulus,in the fourteen day in the stimulus. .Used Enzyme-linked immunosorbent method to determine serum corticosterone in rats after stress. Results:The rats with chronic emotional stress manifested frequent attack,and modify behavior, Weight growth had striking decreased ,The content of cortical ketones in the blood serum had significant decreased, compared withtreatment group and model group :attack and modify behavior had significant increased, the weight growth had significant increased, the content of cortical ketones in the blood serum had significant decreased. Conclusion: The rhizoma anemarrhenae decoction for the improve lever function empty bottle stimulate induced chronic emotional stress for the treatment role.
Key words: rhizoma anemarrhenae, cortical ketones, empty bottle stimulate, Chronic emotional stress
Anemarrhenae is of cool nature, it tastes bitter and sweet,the rolo of clearing heat purging fire and nourishing yin for moistening dryness.Studies have shown that saponins from
Anemarrhenaaspholeloides Bge can reduce the content of cortical ketones in the bloodserum in rats after chronic emotional stress.This experiment usedthe model of empty bottle stimulate to cause chronicemotional stress in rats. From the behavior, weight growth and corticosterone levels, observeand study hizoma anemarrhenaefor the improve lever function empty bottle stimulate induced chronic emotional stress.
1.Materials and Methods
1.1 Instruments and reagents
Anemarrhena,dried rhizome of monocotyledon liliaceae Anemarrhena aspholeloides Bge(Harbin Pharmaceutical GroupCo. Ltd.Shi amedicinedrink, Lotnumber:111023 ), reparation of anemarrhenae water decoction: Take Anemarrhena 100 g, Soak for 1 hour, Plus 20 times theamount of water, decoct 2 times,each time 1.5hours,The two combined filtrate is concentrated to 1.4ml,Rat corticosterone ELISAkits(U.S.R& Dcompany, Lot number:20111201B), Microplatereader(U.S. BIO TEK Corporation, Model:Syner-gy MX);KDC-40 Low-speed centrifuge(HKUST Innovation Co., Ltd. in the best branch).
1.2 Replicated the rats chronic emotional stress model and divded rats into different group
Selected 30 SD rats, detergents, male,weight(200±20)g,(Heilongjiang University of Chinese Medicine Experimental Animal Center, License No. Black move Zi p00701109).The rats was randomly divided into blank group,model group,treatment group,Each group had 10.The rats were feeded one week with single cage to adapt the environment.Trained for drinking water after one week, Contents are: drinked water to rats at 9:00-9:10 am and 9:00-9:10pm. The rest of the time, neither water nor to the bottle.Trained for one week.Drinking Stimulate for 14days after one week dringing training: In addition to the blank group drank water twice a day, The remaining two groups were stimulated in the eighteen day of the training .Given uncertain empty bottle stimulation in the above-mentioned period of water time. Once or twice daily for 14 days.Treatment group began to irrigate stomach anemarrhenae in the second week of stress. Irrigated stomach for 7 days, the dose of2.16g/kg.
1.3 Behavioral observation
Behavioral observation containsaggressive behavior(Attack or bite empty bottles and cages). Exploratory behavior(Visit the occurrence of the water bottle and movement around). Grooming behavior(combing fur and face). Ten minutes of the stress was divided into 10 time periods, Rats were recorded the three actsin each period. Behavior appears denoted 1, otherwise as 0. Observed score within 10 minutes is 0-10 points. Score was based on an average of the results obtained by two observers. One of them was double-blind controlled, The average scores of the last three days during the 14 days traintment were used for statistical analysis.
1.4 Increase of body mass of rats
Weighed each rat by electronic animalscales on the day before water stimulation(Before the experiment ).weighed each rat in the 7th day of stimulation and the 14th day.(the increase of body mass of rats =Current body mass weighing—Body mass before the experiment).Eat freely during the experiment
1.5 Determination of serum corticosterone
After the 14 days stimulus, All rats were orbital blood, Serum was separated,Detected the rat's serum corticosterone levels by Enzyme-linked immunosorbent assay. The standard curve is shown in figure 1.
1.6 Statistics and Data Processing
Used spss13.0 software systems to analyze experimental data. Experimental data expressed as x±s deviation. Use One Way ANOVA, P <0.05 was considered significant difference, P <0.01 was considered to be a very significant difference.
Figure 1. Corticosterone standard curve 2. Results
After the empty bottle stimulation, The behavior of rats in each group, body mass growth, changes in serum corticosterone levels are shown in Table1,2,3. Table1. Behavioral changes in rats compared before and after administration( x±s)
Before administration after administration Attacking exploring grooming attacking exploring grooming
BlankGuoup ModelGroup
0.0±0.0 1.2±0.7 4.2± 1.4 0.0±0.0 1.0±0.5 4.7±1.3 7.1 ± 1.6^ 5.9±1.5^2.9±1.1^ 0.3±0.5 1.6±0.9 3.1 ± 1.1
TreatmengGroup 6.8±1.5^ 6.2± 1.4^2.7± 1.0^ 0.4±0.5 4.3±1.2#
1.2±0.7
Note: compared with blank group AP<0.05, ^P<0.01, compared with model group#p<0.05 Table2 Growth of rat body mass compared( x±s)
Body mass before body mass in the body mass in the
Blank group Model group Treatment group
experiment
225.9± 19.5 232.8± 18.2 230.5 ±17.4
14thday
27.3 ±8.4 18.7±4.6A
21thday
41.2±7.4 32.4 ±6.5A
20.4±4.2A
38.7±5.8#
Note: compared with blank group AP<0.05, compared with model group # p<0.05 Table3 The serum corticosterone levels changes ( x±s)
serum corticosterone levels (ng/mg)
Dose(g/kg). in the 14th day in the 21th day
BlankGroup ModelGroup TreatmengGroup
2.16
42.27 ±11.83 76.62± 12.19^ 71.48± 13.51^
49.18± 14.32 65.31 ± 13.19A 52.01 ± 12.73 #
Note: compared with blank group AP<0.05, ^P<0.01, compared with model group#p<0.05 3.Conclusion
Anemarrhena used to treat fever, the disease often accompanied agitated, anxiety, insomnia and other symptoms, Previous studies that involve fear, anxiety, hopelessness and other negative emotions can cause the release of corticosterone, Studies have confirmed that Timosaponin have antidepressant and reduce corticosterone levels in mice after chronic stress .This experiment uses a chronic emotional stress model that has been proven effective (empety bottole stitulation) .By studying behavior and body weight gain and blood corticosterone levels change in rats with chronic emotional stress found that rhizoma anemarrhenae decoction can significantly reduce the content of
cortical ketones in the blood, Reduce aggressive behavior in rats with stress, Significantly improve this situation that reduced body weight gain in rats. Results showed that therhizoma anemarrhenae for the improve lever function empty bottle stimulateinduced chronic emotional stress for the treatment role.
Corespondent author:
Liting Li, MD, Male, HeilongjiangHarbin, College of Pharmacy, HeilongjiangUniversity of Chinese Medicine, Doctoral Supervisor. Main research interests: Natural drugs and compound biological activity. Email: litingli8888@sohu.com. Foundation item: National Natural Science Foundation of China (81073077)
The Study on the Metabolic Process of Schizandrin in Rats
Tingli LI*, Shengnan LI, Ruixin XU College of Pharmacy of Heilongjiang University of Chinese Medicine,Harbin,150040,China
Abstracts: Research on the metabolic process of the monomer of Schizandrin in rats. Plantthe microdialysis probe into the blood vessel of the rat.Theyare given the monomer of Schizandrin by 20mg kg-1. Use HPLC to detect Schizandrin in dialysate, drawing Mean blood concentration-time curve and calculating the pharmacokinetic parameters of the processes. Absorption and metabolism of the monomer of Schizandrin are very fast. The value of Cmax is high, while t1/2 and MRT are low. The pharmacokinetic parameters are: ke=0.371h-1, ti/2=1.869 h, Tmax=0.75h, Cmax=159.35ug ml" 1, AUCo-t=23L806ug-h-mr\ AUCo.»=282.710ug h ml_1.
The monomer of Schizandrinbelongs to the fast absorption and metabolism type of drug,.
Key word: blood-microdialysis; Schizandrin; metabolism; pharmacokinetic parameters
Schizandrin is one of the main active components of Schisandra chinensis.Its effects of liver-protected and reducing aminopherase in blood are notable.However,there is no report on the metabolic process of Schizandrin in animals.This experiment adopts the method of blood-microdialysis to reserch the metabolic process of the monomer of Schizandrin in rats, then laying the foundation for Pharmacokinetic of Schizandrin.
Materials and methods: Preparation of the blank dialysate: Weigh NaCl 7.137g, KCl0.224g, MgSO40.144g, KH2PO40.054g, NaHCO32.100g and CaCl20.133g was dissolved in 1 L of pure water, prepared the Krebs-RingerVRinger's solution containing NaCl: 122 mmol/L, KCl:3 mmol/L, MgSO4:1.2 mmol/L, KH2PO4:0.4 mmol/L, NaHCO3: 25 mmol/L and CaCl2: 1.2 mmol/L of Ringer's solution, adding 1% Tween-80, suspended for 3 min, sonicated for 30 min. The dialysate should be prepared before using it every once.
Established the analysis methods of Schizandrin by HPLC:
Chromatographic conditions: Column: Hypersil C18 column (4.6 x 250mm, 5p,m); Mobile phase: (methanol: water)= (60:40,V/V); Flow rate:1.0 ml/min; Column temperature:30°C;Detection wavelength: 254nm
Study of Methodology: Preparation of the standard curve; Determination of the LOD and LOQ; Determination Specificity; Study of the intra-day precision; Study of the inter-day precision
Themetabolicprocesses of Schizandrinin rats
Five male Wistar rats (They were provided by the Experimental Animal Center of Heilongjiang University of Chinese Medicine.) weight 300 ± 20g. Feed adaptively one week. Before the experiment, fasting for 12h, and having free access to water. Before the surgery, rats by intra-peritoneal injection of 10% urethane (10ml/kg body weight), rats were anesthetized and injected subcutaneously in saline solution containing heparin (content heparin 60U). Abdominal injection of heparin after 30 min, cutting the fur of rat neck, exposing and separating the right
