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12cortical 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: [email protected]. 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 20mgkg-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, AUC0-t=23L806ug-h-mr\ AUC()-»=282.710ughmr1.
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, MgS040.144g, KH2P040.054g, NaHC032.100g 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, MgS04:1.2 mmol/L, KH2P04:0.4 mmol/L, NaHC03: 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 L0D and L0Q; 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
jugular vein, ligated the distal end, with the ophthalmic scissor cutting a small opening on the surface of the vessel. The cannula of the microdialysis probe should be inserted quickly into the jugular vein of rats, while at the same time, with Tween-80 wet it, and then the blood microdialysis probe is implanted the position of atrial enlargement, which is located at the junction of the superior vena cava and inferior vena cava (from the junctionof The external jugular vein and internal jugular vein approximately 33-34mm, Fig. 1). The probes and the vascular were fixed together by non-adsorbing surgical suture, in order to prevent the probes slipping. After surgery, the wound covering gauze wetted with saline. Turning on the CMA microdialysis perfusion pump, with a blank dialysate perfusion 1h, perfusion flow rate of 1ul/min. Orally administered to rats Schizandrin 20mg/kg (which is dissolved by 3% Tween-80 solution), turn on microdialysis automatic collection device immediately after administration, collected one sample every 30min, collected nine samples, with the blank dialysate continued perfusion 2h, then turned to a schizandrin concentration of 0.1mg/ml of dialysate perfusion, balance 1 h, collecting three consecutive samples, detection of the contentof schisandrin in collected samples by HPLC.Calculating determination of the probe of vivo recovery, the experimental data of each animal corrected by their ownvivo recovery. Determination of the vivo recovery of the formula: R=[1-Cout/Cin] x 100% The rats were sacrificed by thoracotomy, check the correct position of the probe implantation, eliminating the data of the incorrect positions. All statistical evaluations were performed with 3P97 pharmacokinetic software, calculating the various moments of pharmacokinetic parameters and drawingmean blood concentration - time curve.
Fig. 1 The location of the implantedblood
W ■ , : dialysate j- i • u
microdialysis probe
jugulaiVuL vein
axillary c^JsA. vein
microdialysis probe_
Results and discussion 1. Results
1.1 Results of the methodological research 1.1.1 Preparation of the standard curve
Linear regression of its concentration in the peak area of schisandrin, the linear regression equation is: Y=5012.3X-7984.3, R2=0.9993, schisandrin in the concentration of 0.2-1000 ug in concentration range, showed a good linear relationship. (Table 2, Fig. 2)
Concentration C (ug/ml) 0.2 2 20 100 200 500 1000
Peak area (A) 1689 10535 107346 501685 1009507 2395041 5051685
Table 2 the Standard curve table of peak area-concentration
6000000 5000000 4000000
IS 3000000 §
^ 2000000 & 1000000 0
-1000000
0
5012.3x - 7984.3
R2
0.9993
200 400 600 800 1000 1200 Schizandrin concentration C(ug/mL)
Fig. 2 the standard curvediagram of peak area-concentration
1.1.2 LOD andLOQ
In accordance with the signal to noise ratio (S/N) of 3:1 meter schizandrol determine a detection limit of 0.05 ug/ml; according to noise ratio (S/N) of 10:1 meter to determine the limit of quantification schizandrin of 0.2 ug/ml.
1.1.3 Specificity Specificity result in Fig. 3, Fig. 4
A
0.501-
0.45" 0.4Ö-;
Ö.30-< 0.25-
2.00 4.00 e.oo 6.00 io!oo iz!m 14.00 10.00 is'oo 2o!oo 2e!oo
B
Fig. 3 the HPLC chromatogram of the Schizandrin and the blank dialysate A: Schizandrin;B:Blank dialysate
:7ig.4the HPLC chromatogram of the blank dialysate of containing Schizandrin
1.1.4 the intra-day precision and the interday precision (Table 3 , Table 4)
Concentratio n (C) Peak area (A) X RSD%
1000 ug/ml 5048165 5074531 5065934 5025391 5003194 5043443 0.58
200 ug/ml 1020832 1010394 996489 1035642 1039889 1020649 1.76
20 ug/ml 103346 99753 101556 102844 105897 102679 2.21
Table 3 the intra-day precision
The results showed that: high, medium and low concentrations of chromatographic peak ar
RSD% values between 0.58% ~ 2.21%, less than 3.0%, in line with the quantitative requirements.
Concentratio Peak area (A) v RSD%
n (C) 1d 2d 3d 4d 5d X
1000 ug/ml 5048165 5098462 5026983 5016854 5012697 5040632 0.69
200 ug/ml 1020832 998725 987543 1001299 1036981 1009076 1.94
20 ug/ml 103346 998264 102368 989315 106372 102176 2.89
Table 4the interday precision
The results showed that: high, medium and low concentrations 5d consecutive chromatographic peak area of RSD% values between 0.69% ~ 2.89%, less than 3.0%, in line with the quantitative requirements.
1.1.5 Mean blood concentration-time curve
The experimental results showed that: schizandrol monomer after administration, plasma concentrations of rats immediately elevated concentrations peaked in within 1h, 2.5h after drug-time curve to flatten. 3.75 h after the plasma concentration decreased to 25ug/ml or less. (Table 5, Fig. 5)
-
Jrj'M---
nr
0 1 2 3- -i &
i(h)
Fig. 5 Mean blood concentration-time curve of Schizandrin after oral administration to rats 1. Discussion
Fructus Schisandrae Chinensis is the dried ripe fruit of Magnoliaceae Schisandra chinensis (Turcz.) Baill, and Sphenanthera Rehd. et Wils, the former commonly known as Schisandra, the latter commonly known as Kadsura. Schisandra is one of the most commonly used herbs in traditional Chinese medicine, research shows that Schisandra has a clear role in protecting the liver, including lipid peroxidation, protecting liver cell membrane and promoting the metabolism of toxic substances.The 10th edition Chinese Pharmacopoeia sets Schisandrin is the index component of traditional Chinese medicine in Hugan tablets, the experiment proved schizandrol hepatoprotective effect is remarkable.
Table 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:
