Studies On The HPLC Fingerprint Of JieDuYiHao And The Effects Of JieDuYiHao On Methamphetamine Induced Behavioral Sensitization In Mice

Objective: Drug addiction is a chronic,recurrent brain disease that is characterized by a continuous,compulsive use of dependent drugs for non-medical purposes.Methamphetamine(METH)is a kind of amphetamine-type central stimulant,which is seriously harmful to human health,and brings new problems and challenges to narcotics control in China.The mechanism of METH addiction is complex.The multitargeted Chinese herbal compounds are expected to be a potential therapeutic drug of METH addiction.Through our previous experimental screening,we prepared a drug JDYH consisting of 8 Chinese herbs.A series of quality control and safety evaluation experiments were conducted to ensure the stability and safety of JDYH.We also investigated the effect of JDYH on METH induced behavioral sensitization in mice to find a promising anticraving candidate for the management of METH addiction.Methods: 1.Preparation of JDYH.JDYH consists of the following components:Corydalis yanhusuo.W.T.Wang,Poria Cocos,Codonopsis pilosula(Franch)Nannf,Glycyrrhiza uralensis Fisch,Atractylodes macrocephala koidz,Astragalus membranaceus(Fisch)Bunge,Panax quinquefolius L,Angelica Sinersis.The drug weighing 94 g was soaked in 1400 ml distilled water for 30 min.The liquid was decocted to 400 ml with a traditional Chinese medicine decoction,then the residue was filtered out,and the JDYH water decoction was obtained.2.Quality testing experiment.The high performance liquid chromatography(HPLC)fingerprint was conducted with the Ultimate TM XB-C18(250 mm × 4.6 mm)column in gradient elution.The analysis of Corydaline,Tetrahydropalmatine,Berberine and Glycyrrhizic acid was conducted with the mobile phase consisted of acetonitrile-water-phosphoric acid,at column temperature 30 ?,flow rate of 1.0 mL/min and detective wavelengths of 280 nm.The analysis of Ginsenoside Re and Ginsenoside Rb1 was conducted with the mobile phase consisted of acetonitrile-water,at column temperature 30 ?,flow rate of 1.0 mL/min and detective wavelengths of 203 nm.3.Acute and long-term toxicity test.To observe the toxic effect of JDYH,body weight,food intake,water intake,and biochemical indexes of SD rats were detected in acute and 3 months long-term toxicity test,which provided a basis for clinical safe drug use.4.Locomotor activity test.Locomotor activity was detected after intragastrical adminstration with JDYH for 1 day or 7 days in mice.After 7 days of treatment with JDYH,mice were given 7 days of withdrawal.Locomotor activity was detected after the withdrawal time in mice.The locomotor activity test was conducted to evaluate the effect of JDYH on the locomotor activity and whether JDYH itself would produce behavioral sensitization.5.High activity test.Locomotor activity was detected to observe the effect of JDYH on the hyperactivity of mice with single METH exposure.6.Behavioral sensitization test.The mouse model of behavioral sensitization was established by repeated discontinuous METH treatment,and JDYH was given in the three stages of behavioral sensitization to observe the effects of JDYH on the development,transfer and expression of behavioral sensitization induced by METH.Result: 1.The HPLC Fingerprint of JDYH showed that the concentrations of Corydaline,Tetrathydropalamatine,Berberine were 28.33 ?g/ml,27.66 ?g/ml,3.18 ?g/ml.The fingerprint of ten batches of JDYH decoction were established,which all contained 19 common peaks,the similarities among different batches were above 0.998,with RSD <1%.2.In the acute toxicity test,no toxicity reaction occurred in rats treated with JDYH at67.68 g/kg which was equivalent to 80 times of adult clinical dosage.In the long-term toxicity test,no changes were observed in body weight,food intake,water intake,clinical biochemistry,gross necropsy and histopathology.3.In the locomotor activity test,compared with the control group,there was no significant difference in the locomotor activity of mice treated with low-dose(14.12 g/kg)or high-dose(56.48 g/kg)of JDYH.4.In the high activity test,compared with the control group,the hyperactivity induced by acute METH was not reduced by JDYH.5.In the behavioral sensitization test,compared with the model group,the development,transfer and expression of MA-induced sensitization were significant suppressed by JDYH.Conclusion: 1.According to the HPLC fingerprint of 10 batches JDYH sample,the relative retention time of different batches of JDYH was basically the same,and the composition was basically the same,indicating that the quality of each batch of JDYH was stable.2.JDYH had no significant toxicity in acute toxicity test and long-term toxicity test,indicating that JDYH was safe to be used in clinical dosage.3.Compared with the control group,no significant difference was observed in the locomotor activity of the mice in JDYH low or high-dose group,indicating that JDYH had no effect on locomotor activity in mice and did not produce behavioral sensitized effect.4.Single JDYH administration had no effect on hyperactivity induced by METH in mice.5.JDYH could inhibit the development,transfer and expression of METH-induced sensitization in mice,suggesting that JDYH had therapeutic effect on METH-induced psychic dependence.