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5Table 5 pharmacokinetic parameters of Schizandrin after oral administration to rats
Parameters x ±s
ke/h-1 0.371±0.050
t1/2/h 1.869±0.241
Tmax/h 0.750±0.000
Cmax/ug ■ ml-1 159.35±31.70
AUCO-t/ug ■ h ■ ml-1 231.806±15.806
AUCO-^/ug ■ h ■ ml-1 282.710±16.538
Nowadays, with the development of molecular biology and the continuous improvement of detection methods, herbal pharmacokinetic study has been greatly developed and has become an important field of Chinese medicine research. Chinese medicine pharmacokinetic studies have significance to elucidate the mechanism of action ,to reveal the herbs and its scientific content, to design and select dosing regimens,topromote the development of new drugs and dosage forms of Chinese medicine to improve and enhance the level of quality, promoting Chinese medicine to the world, and ultimately to achieve modernization of Chinese medicine.
In recent years,microdialysis technique developed a new type of bio-sampling technology, there isthe outstanding advantages ,such as in vivo, real-time, online etc.
In this study, microdialysis technique study blood schizandrol metabolism in rats in vivo.
By schisandrin monomer in rat plasma concentration-time curve can be seen, schisandrin monomer in rats after intragastric administration, drug concentration in blood increased immediately, concentrations peaked within 1 h, 2.5 h after the concentration-time curve tends to be gentle, after 3.75 h blood concentration decreased to 25 ug/ml the following. Pharmacokinetic parameters showed that schisandrin absorbed more rapidly in rats, the peak at higher blood drug concentration, the shorter half-life period in vivo, mean residence time is short, quick absorption, quick metabolism of drugs, this agreement with previous reports.
In summary, this study illustrate the monomer components of traditional Chinese medicine metabolic processes in the body, and thento study the clinical medicine with schisandrin as the index components, like HUGAN tablets in the body pharmacokinetic behavior provide research paradigm and lay the preliminary foundation.
Corespondent author:
Liting Li, MD, Male, Heilongjiang Harbin, College of Pharmacy, Heilongjiang University of Chinese Medicine, Doctoral Supervisor. Main research interests: Natural drugs and compound biological activity. Email: [email protected]. Foundation item: National Natural Science Foundation of China (30672634)
The Effect of Different Illumination Condition on The Content Change of 5-HT and DA In 7-Day-Old Female Drosophila's Brain
Liu Siqi, Bian Hongsheng, Jin Yang and Li Tingli
(Heilongjiang University of Chinese Medicine, Harbin 150040, China)
Abstracts
Objective: The goal of the present study is to investigate the effect of different illumination condition on the content change of 5-HT and DA in 7-day-old female drosophila's brain. Methods:
Choosing 7-day-old female drosophila as the object of observation, the dissertation observes the content change of 5-HT and DA in drosophila's brain in different time(16:00, 19:00, 22:00, 1:00, 4:00 and 7:00) in 12/12h alternating light and dark and 24h constant darkness environment and investigates its effect. Results: With the deepening of sleep, the content of 5-HT and DA in 7-day-old female drosophila's brain first decreases, then increases and decreases again in 12/12h alternating light and dark environment. While the content of 5-HT and DA in female drosophila's brain firstly increases and then decreases in 24h constant darkness environment, and the content of 5-HT decreases significantly at 16:00 in 12/12h alternating light and dark environment (*p<0.05). The content of DA increases gradually and the content of DA in female drosophila's brain at 16:00, 19:00, 22:00 greatly decreases compared with it in the 12/12h alternating light and dark environment at the same time (*p<0.05). However, the gap between them shortens gradually. After 1 o'clock, the content of female flies' DA in 24h constant darkness environment increases more than DA in 12/12h alternating light and dark environment (*p<0.05). Conclusion: The content of 7-day-old CS female drosophila's 5-HT and DA will change as time changes in 12/12h alternating light and dark environment and 24h constant light environment.
Key words: Drosophila, 5-HT, DA
Introduction
In recent years, Drosophila has been defined as a model organism by domestic and foreign scientists for sleep studies[1'2]. Based on previous studies, A circadian rhythm of sleep inDrosophila was found, and the related factors of age and sex could affect the sleep time of Drosophila [3]. What is it look like the change of 5-HT and DA of drosophila by light effects? Only recently has interest in this change led to research on Drosophila.Choosing 7-day-old female flies as the object of observation, the dissertation observes the content change of 5-HT and DA in female flies' brain in different time (16:00, 19:00, 22:00, 1:00, 4:00 and 7:00) in 12/12h alternating light and dark and 24h constant darkness environment and investigates its effect.
Materials and methods
The following methods of investigation were used the Wild-Type Canton S Flies (Drosophila melanogaster) provided by the College of Life Sciences, PekingUniversity.
Experimental Instrument Multifunctional microplate reader (Biotek,Model: SynergyMx) Experimental reagents Fruit fly DA ELISA Kit (from RB, Lot: 20120301B)Fruit fly 5-HT ELISA Kit (from RB, Lot: 20120302B) Experimental temperature: 24 ± 1°C; Humidity: 50 ~ 60%; Light: 12h light-dark auto cycle (7:00 lights, turn off the lights 19:00).Experimental mediumMaize culture medium was used in this study. Culture medium prescription: Distilled water 460g, Maizena 42g, Cane sugar 31g, Agar 3g, Yeast powder 3g, Propanoic acid 3ml.
Determination of Enzyme-linked immunosorbent assay(ELISA)
©From room temperature equilibrium after 20min of foil pouch microtiter plates, corresponds to the order in accordance with the standards were added in 50ul blank microplate. ©Setting the standard holes and sample wells, standard holes each plus different concentrations of standard 50ul. ©Adding the test sample 10ul and sample dilution 40ul in sample holes. Blank wells nothing. ©Except for blank wells,adding100ul horseradish peroxidase (HRP)labeled detection antibody in every standard holes and sample holes, closure plate membrane sealed reaction hole.©Incubating reaction 60 min at 37°. ©Throw away the liquid, pouring washing solution into every hole, standing 10 min, rejection to washing solution. Patting dry with absorbent paper. washing holes with washer 5 times, standing 10-20 sec. every times. ©Each hole by adding substrate A, B mixture of 50ul. ©Incubating reaction without bright about 60 min at 37°.©Adding 50ul of Stop Solution to each well, the reaction was terminated. Results to determine©Instrument values: At a wavelength of 450nm on a microplate reader to read the OD value of each well.©In OD ordinate, with standard concentration as abscissa, draw a standard
regression curve.©According to OD values of samples to find the corresponding concentration range.®Sensitivity:1.0 pg/ml.
Drosophila brain 5-HT and DA Preparation of test sample Remove from the liquid nitrogen frozen pipes with Drosophila heads, The Drosophila heads move within homogenizer, Adding 0.5mL saline homogenate, Centrifuge freezing (TOMOS,3-18R,USA) 3000R/Min 10min, 200ul supernatant in labeled tubes.
Group of Experiments Collected unmated female flies within 8h of emergence, flies shallow anesthesia with CO2. Aliquot the flies within the glass tube, group(B) were randomly divided into 12/12h alternating light and dark, group(M) were randomly divided into 24h continuous darkness. Group(B) flies are placed in the finger tube with basal medium, feeding 7 days in12/12h alternating light and dark, group(M)flies are placed in the finger tube with basal medium, feeding 7 days in 24h continuous darkness. Taking the flies heads at 6:00, 19:00, 22:00 and next day 1:00, 4:00, 7:00 (N=5 samples, each sample 30). Follow the simple random sampling method five samples from each group were randomly selected three parallel samples and set aside. Data processing and statistics The experimental results are x±s , using SPSS17.0 statistical software, when comparing more than two groups using single factor analysis of variance for data processing, LSD method using pairwise comparison, comparisons between the two groups using independent sample t-test. Standard of statistical significance is p<0.05.
Results and discussion
Drosophila melanogasterwhich lay the foundation of classical genetics is the most commonly used in Drosophila genetics and one of the important model organisms. At present,Drosophila Activity Monitor with video recording method is used in the study of the sleep of the flies. The method can monitor a large number of flies in the meantime and can be used in the study of screening of Drosophila. More than 60% of human disease genes can be found in the Drosophila [4' 5].Therefore, scientists first study the physiological mechanisms of some lower organisms. Then results of their research are applied to higher organisms and humans. Table 1 7-day-old female flies brain 5-HT content at different time ( x±s)_
Group Time Number samples of 5-HT content <ng/ml)
12/12h alternating light and dark 16:00 30 140.16±24.13
24h constant darkness 16:00 30 68.73±19.64*
12/12h alternating light and dark 19:00 30 114.37±43.77
24h constant darkness 19:00 30 90.16±26.37
12/12h alternating light and dark 22:00 30 148.09±5.05
24h constant darkness 22:00 30 120.32±25.25
12/12h alternating light and dark 1:00 30 82.22±24.13
24h constant darkness 1:00 30 102.86±19.64
12/12h alternating light and dark 4:00 30 100.07±35.91
24h constant darkness 4:00 30 110.79±20.76
12/12h alternating light and dark 7:00 30 66.35±10.66
24h constant darkness 7:00 30 73.89±5.61
Note:The same time point 24h continuous darkness group compared with 12/12h alternating light and dark:*p<0.05, **p<0.0112/12h alternating light and dark environments adjacent time point compared two groups: Ap<0.05, AAp<0.01; 24h under continuous darkness environments adjacent time point compared two groups:: #p<0.05, ##p<0.01.
The experimental study found, in continuous 24h dark and 12/12h alternating light and dark environments female Drosophila brain 5-HT and DA levels will change with time (table1, table2). Around 19:00, the content of DA and 5-HT in female drosophila's brain slightly lower, but still much higher than in continuous 24h darkness environment female flies' brain 5-HT and DA levels. And combined with previous research findings: in 12/12h alternating light and dark environments,
around 19:00 flies occurs peak activity; the environment in 24h continuous darkness flies peak activity disappear. This phenomenon reminds us of Drosophila activity and sleep rhythms may be associated with brain 5-HT and DA content. Existing 5-HT has been reported to promote the role of sleep[6'7], and DA to promote animals related to the role of awakening[89], the experiment with them minded.
Conclusion
The content of 7-day-old CS female drosophila's 5-HT and DA will change as time changes in 12/12h alternating light and dark environment and 24h constant light environment. Experimental results show:
With the depth of sleep, in 12/12h alternating light and dark environments 7-day-old female Drosophila brain 5-HT content decreased first, then increased, then decreased,While the content of 5-HT in female drosophila's brain firstly increases and then decreases in 24h constant darkness environment.And the content of 5-HT in drosophila's brain,continuous 24h darkness group were lower than 5-HT of 12/12h alternating light and dark at 16:00 (*p<0.05) .
Table2 7-day-old female flies brain DA content at different time ( x±s)
Group Time Number of samples DA content Cng/ml)
12/12h alternating light and dark 16:00 30 121.28±68.55
24h constant darkness 16:00 30 17.62±9.05*
12/12h alternating light and dark 19:00 30 87.43±23.28
24h constant darkness 19:00 30 25.55±15.09*
12/12h alternating light and dark 22:00 30 111.22±6.04
24h constant darkness 22:00 30 57.26±14.23*
12/12h alternating light and dark 1:00 30 62.74±32.34
24h constant darkness 1:00 30 68.54±17.25
12/12h alternating light and dark 4:00 30 56.95±3.45
24h constant darkness 4:00 30 83.17±6.89*
12/12h alternating light and dark 7:00 30 34.39±31.91
24h constant darkness 7:00 30 111.83±47.43*
Note:The same time point 24h continuous darkness group compared with 12/12h alternating light and dark:*p<0.05,**p<0.01; 12/12h alternating light and dark environments adjacent time point compared two groups: Ap<0.05, AAp<0.01; 24h under continuous darkness environments adjacent time point compared two groups:: #p<0.05, ##p<0.01. Experimental results show:
With the depth of sleep, in 12/12h alternating light and dark environments 7-day-old female flies brain DA content decreased first, then increased, then decreased,While the content of DA in female flies' brain gradually increases in 24h constant darkness environment.In 24h continuous darkness 16:00,19:00,22:00 female flies brain DA content lower than the same time in 12/12h alternating light and dark environments (*p<0.05) ,however, the gap between the two decreased.After 1:00,the content of female flies' DA in 24h constant darkness environment increases more than it in 12/12h alternating light and dark environment(*p<0.05). References
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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: [email protected]. Foundation item: National Natural Science Foundation of China (81073077)
Based on sunflower protect liver slices of CCL4 hepatic injury model descending transaminase function and its mechanism of action research
Wang Qi, Zhao Hong Hai, Li Tingli HeiLongJiang University Of Chinese Medicine, Harbin, China
Abstracts:To study the Sunflower brand liver tablets down the role of the characteristics of aminotransferase, and to explore the mechanism of lowering transaminase.
Key words:Sunflower Brand liver tablets, Transaminase, Mechanism, Acute liver injury
CCl4
Introduction:
Liver disease is a major disease threat to human health at present,all kinds of liver diseases,Liver damage and shall be pathological phenomena.Evaluation or development of drug effect that protect liver, has always been a hot spot in the study of liver disease drug.Though various kinds of liver disease drug emerge in endlessly, but there is no better specific can obviously protect liver, reduce injuries and promote the regeneration of the liver cells.Existing drug treatment also is pure treatment for symptoms, more can't achieve the goal of completely cured.
Protect liver parenchyma cells and reduce liver damage can obviously reduce the recession of liver function, can even reverse the disease development. However, because of sunflower protect liver slices and protect liver transaminase function of medicinal material uncertainty, unclear mechanism is not clear, the metabolic process, the low level of quality control standards, thus affecting the curative effect of the sunflower protect liver slices, difficult to get the international drug market recognition, lack of competitiveness in international market. This paper conducted a sunflower protect liver slices descending transaminase function and its mechanism of action research, aims to elucidate its prevention and treatment of liver damage, lowering transaminase function characteristics, and discussed its mechanism of action of descending transaminase. Guided
