ABSTRACT
Objective: To develop an accurate and effective method to identify the original animals of Chinese medicine Pheretima (Dilong in Chinese) and its adulterants by using mitochondrial genes, cytochrome coxidase I (CO I) and 16 S rRNA as DNA barcoding sequences to certify and improve morphological analysis. Methods: A total of 66 samples were initially identified according to their morphological characteristics, and then their CO I and 16 S rRNA sequences were simultaneously amplified for DNA barcoding. The two gene sequences were amplified with improved primers. One-step double PCR was used and the experimental conditions were optimized. The genetic distance among and within species of Pheretima and its hybrids was calculated by MEGA 5.1. The N-J tree was constructed based on K2P model. Results: Combined with morphological identification, CO I and 16S rRNA double DNA barcoding identification can accurately identify Pheretima and its adulterants. Conclusion: Morphological identification is the basis of molecular identification. Molecular identification can be a powerful supplement to the morphological identification. The combination of molecular identification and morphological identification can increase the accuracy of the identification of the original animals of Pheretima and its adulterants. Double DNA barcoding can also provide a solution for the identification of the medicinal materials of Pheretima and other animal medicinal materials.
ABSTRACT
Objective To observe the change of dopamine receptor in ventral tegmental area (VTA), nucleus accumbens (NAc) and prefrontal cortex (PFC) in morphine-dependent rats, and the regulation effect of electroacupuncture (EA). Method Thirty male SD rats were randomized into a control group, a model group and an EA group, 10 rats each. Morphine-dependent rat models were induced by morphine self-administration. In the EA group, bilateral Jiaji points (EX-B2) of T5 and L2 were selected, and the EA intervention lasted 4 d. Western blotting method was adopted to observe the change of the contents of dopamine receptor D1 and D2 in VTA, NAc and PFC of the rats. Result After the intervention, compared to the model group, the morphine intake was reduced significantly in the EA group (P<0.05); the level of D2 declined significantly in VTA of rats in the model group (P<0.05); the level of D1 increased significantly and D2 declined significantly in NAc of rats in the model group (P<0.05); the level of D1 declined significantly and D2 increased significantly in PFC of rats in the model group (P<0.05); EA produced regulation effect on the altered contents of D1 and D2 in the cerebral areas mentioned above, approaching the normal level. Conclusion EA can inhibit the hunger of addiction rats for morphine to some extent; the contents of dopamine receptors in dopamine projection pathway will take adaptive changes after morphine addiction, while EA can regulate the abnormal expressions of dopamine receptors, producing a protective effect on dopamine receptors of morphine-dependent rats.