Oryzanol and vitamin B1 enhance the anxiolytic effect of diazepam

[ Abstract ] Objective : To study the anti-anxiety effect of diazepam, oryzanol and vitamin B1 in a ratio of 1 : 32 : 8 , and compare with diazepam. METHODS : The model of the large and mouse elevated cross mystery model and the rat pentamethine tetrazolium discrimination effect were used. Results: In rats and mice in elevated plus-maze, after diazepam in combination with vitamin B1 and oryzanol, and mice in the open arm was significantly increased percentage of the residence time, it showed significant anxiolytic effects; and its arm in the open the percentage increase in residence time, significantly higher than its effect alone equal levels of diazepam. Pentaerythritol tetrazolium

In the discrimination effect , the ED50 value of the discrimination effect against rat penta-tetrazole after the combination was 23. 4 mg · kg

That diazepam content of 0. 6mg · kg, far lower than the ED50 values diazepam unilateral 3. 5mg · kg. Conclusion : Oryzanol and vitamin B1 have an anxiolytic effect of diazepam.

[ Key words ] diazepam ; oryzanol ; vitamin B1; elevated plus maze ; pentamethine tetrazolium discrimination experiment

    Diazepam is a more commonly used anti-anxiety drug , but it will be dependent after long-term use , so how to reduce its dependence while ensuring clinical use is the first consideration of such drugs. Synergy between the case of using a pharmaceutical, by combination with other drugs, to improve its efficacy, so that produce the same efficacy, can greatly reduce the amount of use of these drugs, thus reducing thermal dependency thereof, which is to reduce Dependent drug dependence is very effective

a way. In this study, diazepam, oryzanol and vitamin B1 of 1: 32: 8 ratio composition of Compound (hereinafter referred to as valley Vivian, in another paper published in the prescription basis) study the anxiolytic effect after, found that the combination generating the same In the case of an anxiolytic effect , the amount of diazepam can be greatly reduced.

Materials and Method

1 drug

Dixi ( Datong Huida Pharmaceutical Factory , batch number : 960610), oryzanol , vitamin B1, batch number : 960411, grain and oil from Jining City , Shandong Province

Chemical plants, formulated as suspension in 1% carboxymethylcellulose. Pentamethyltetrazolium , a Sigma product , formulated with a physiological sodium chloride solution.

2 animals

Kunming mice , male , 18 ~ 25g; Wistar rats , male , 180 ~ 250g . The experimental animals were provided by the Experimental Animal Department of Peking University Medical School , which met the national second-level experimental animal standards.

3 methods

3.1 mouse elevated cross puzzle experiment

The labyrinth is made of wood and consists of 2 open arms and 2 closed arms. Open arms of 30cm × 5cm, closed arms of 30 cm × 5cm × 15cm, the open arms and closed arms at the intersection of a 5cm × 5cm of the central platform, the bottom of the open arms and closed arms from the ground 38. 5cm. Experiments in the morning 8: 30: 30 ~ 11. After gently caught maze experimental group and control group animals were orally administered drug or solvent central 30min, the animal's head towards the open arms, immediately after the recording start release the hand within 5min animals staying in open arms and closed arms Time and number of entries , and calculate the open arm staying time in the open arm and closed

The percentage of arm stay time. 3.2 Rat elevated cross puzzle experiment labyrinth is wooden ,

It consists of 2 open arms and 2 closed arms. The open arm is 50cm × 10cm, the closed arm is 50cm × 10cm × 40cm, the open arm and the closed arm are at the center of a 10cm × 10cm platform , and the open arm and the bottom of the closed arm are 50cm away from the ground . Experiments in the morning 8: 30: 30 ~ 11. After gently caught maze experimental group and control group animals were orally administered drug or solvent central 30min, the animal's head towards the open arms, immediately after the recording start release the hand within 5min animals staying in open arms and closed arms Time and number of entries , and calculate the open arm staying time in the open arm and closed

The percentage of arm stay time.

3. 3 penta-tetrazole discrimination antagonism experiment

3.3. 1 Drug Identification Device The rat identification training box is an improved Skiner -type double-bar training box. The training box is placed in a control box with light, sound and ventilation. The experimental process is completed automatically by the computer.

3.3. 2 Establish a rat pentamethine tetrazolium discrimination model. One rod in the training box is a drug ( pentamethene tetrazolium ) rod , and the other rod is a saline rod . Rats are intraperitoneally injected with pentamethine tetrazolium every day. After 20 minutes of azole 20mg · kg-1 (D phase ) or physiological sodium chloride solution (N phase ) , put it into the training box and start training the rats to step on the drug rod in the D phase and step on the saline rod in the N phase. When you step on the correct rod, you can get food fortification , and you ca n't strengthen the step bar. From FRl (a right pedal lever to give a reinforced) starts, by D, N, D, N. .. The order of circulation, a daily training 30min experimental period, the correct recording of rat foot bar at the end of each experiment. When the experiment of 4 consecutive correct pedal lever rate greater than 80%, the FR increases 1 until FR10 (right pedal lever 10 to give a reinforced). Under the condition of FR10 , the correct rate of continuous 4d pedals is above 80% , indicating that the pentamethene has been established in rats.

The discriminating effect of tetrazolium.

3.3. 3 penta-tetrazole discrimination effect against the experiment Rats with stable discrimination effect on penta-tetrazole were intraperitoneally injected with 20 mg · kg-1 of pentamethine 15 min after intragastric administration of the test drug or solvent. Tetrazolium , 15 min later , was placed in a discrimination stimulation chamber to determine the discrimination of pentaerythritol in rats.

4 data processing

Data were expressed as mean ± standard deviation , group t- test was used for comparison between groups , and ED50 value was calculated by Bliss method.

Result

1 mouse elevated plus maze test anxiolytic compared with the control group, the percentage of the valley Vivian stay in the open arms time and three doses of diazepam three doses were significantly increased (P <0. 01), it indicates that have had Significant anxiolytic effects. Diazepam large, medium and small doses of three dose groups, respectively, equal to the amount corresponding dose of diazepam Valley Vian contained. Valley Vian compared to the corresponding dose of diazepam, after Valley Vian high-dose group in the percentage of open arms residence time was significantly higher than diazepam high-dose group, indicating that diazepam and oryzanol, vitamin B1 combined, diazepam anti- The anxiety effect is significantly enhanced.

2 anxiolytic rat elevated plus maze test compared with the control group, the percentage of the valley Vivian stay in the open arms time and three doses of diazepam three doses were significantly increased (P <0. 01), it indicates that have had Significant anxiolytic effects. Diazepam dose three dose groups respectively corresponding dose valley Vian equal amounts of diazepam group contained therein. Valley Vian compared to the corresponding dose of diazepam, Valley Vian large, medium-dose group in the percentage of open arms residence time was significantly higher than large diazepam dose group, indicating that diazepam and oryzanol, vitamin B1 combined after the The anti-anxiety effect of Xiqiao has been significantly enhanced. 3 penta-tetrazole discrimination effect against Shigian Wei'an and diazepam against rat penta-tetrazole

The ED50 values ​​of the stimulating effects were 23. 4 and 3.5 mg · kg- 1 , respectively . The dose of compound diazepam 23. 4 mg · kg-1 contained 0.2 mg · kg-1 of diazepam , which was much lower than the ED50 value measured by diazepam alone 3. 5 mg · kg- 1

. This suggests that diazepam combined with oryzanol and vitamin B1, the anxiolytic effects of diazepam has been significantly strengthened.

Discuss

In this study, three anti-anxiety methods were used : the large and mouse elevated cross mystery method and the rat pentamethine tetrazolium discrimination method . The anti-anxiety effect of diazepam with oryzanol and vitamin B1 was studied . And compared with the anti-anxiety effect of diazepam alone. Three methods have shown that anxiolytic effects of diazepam after the combination has been significantly strengthened, but its mechanism is not fully understood. The synergistic effect of oryzanol is likely to be related to its regulation of diencephalic function, activation of the hypothalamic and limbic systems associated with the autonomic nervous system, and improvement of autonomic function ; the synergistic effect of vitamin B1 is likely to be related to its role in energy metabolism. After diazepam combined with oryzanol and vitamin B1, greatly enhancing the anxiolytic effects of diazepam, which is clinically very meaningful.

Because in the case of the same pharmacodynamics , this combination can reduce the amount of diazepam , thereby reducing its dependence.