| ObjectiveThe experimental study could provide new basis that the treatment of electro-acupuncture and catgut embedding is useful for the clinical patient suffering from drug tolerance, by comparing the influence of electro-acupuncture and catgut embedding regulate abnormal circadian rhythm and some behavioral tests,and the change of expression of Perl gene and Per2 gene in suprachiasmatic nucleus(SCN) of mice bearing morphine tolerance after electro-acupuncture or catgut embedding.Methods32 C57BL/6J male selected and domesticated mice, were randomly divided into Control group(C,n=8), Model group(M,n=8),Electro-acupuncture group(EA,n=8) and Catgut embedding group(CE,n=8).While the mice of C group were given 10ml/kg isotonic saline under the skin on 8am and 8pm,the M group,EA group and CE group mice were built the Morphine-tolerance models by injecting 10mg/kg Morphine Hydrochloride Injection twice a day for 7 days consecutively with I.h.After injected drug in the first morning,the EA group began to receive treatment by using the HANS Stimulator(HANS-2000) to administer electro-acupuncture at bilateral "Shenshu" point (BL23) for 7 days consecutively (standard:0.2mA,2/15Hz,15min once a day). At the same time,the mice of CE group were implanted catgut at the bilateral piont of "shenshu" in the first day after using morphine.After the injection of morphine or saline for 30 minutes,we measured the tail flick latency as an index of pain threshold in the firs,third,5th,7th day. Animals were sacrificed respectively in the morning of the 7th day.then we collected the samples of SCN in mice and measured the the expression of Perl gene and Per2 gene in SCN of mice during RT-PCR method.Results1. Behavioral results:1). The pain threshold test:There were no significant differences (P>0.05) about primary pain threshold among four groups.And there were no significant differences (P>0.05) both the pain threshold in the first,third,5th,7th day and primary in the C group. There were significant differences (P<0.05) both the pain threshold of the EA group, the CE group and the M group in the first,third,5th day and primary. In the 7th day, the M group’s pain threshold had no significant differences (P>0.05) during comparing with its primary pain threshold. And the EA group, the CE group had significant differences (P<0.05) during comparing with their primary pain threshold. There were no significant differences both the EA group and the CE group(P>0.05).2). Weight:There were no significant differences (P>0.05) in four groups. Since the morphine-tolerance modle started, the control group was normal (P>0.05) Others had similar decreased trends about weight during the 7 days. Compared with the C group, Each group was significant differences (P<0.05). It was no differences between the EA group and CE group(P>0.05), but compared with the M group(P<0.05).2. The circadian rhythm in 7 days:There were no significant differences about the activity, amplitude and period among all of the groups (P>0.05).When the recorder started work,All groups mice were not abnormal in period as usual(P>0.05). The EA group, M group, and CE group had significant differences in amplitude and activity compared with the C group(P<0.05). There also had significant differences between EA, CE and M in amplitude(P<0.05), But had no change in activity and amplitude both EA group and CE group,in activity among three groups.3. The expression levels of perl and per2 in SCN:We found that the expression levels of Perl and Per2 in the EA group, M group, and CE group had decline compared with the C group (P<0.05). And it had significant differences between the EA group and the M group(P<0.05). Compared with the M group, the expression levels of Perl and Per2 in the CE group were up-regulated,but This trend was not statistically significant(P>0.05).Conclusion:1. The Weight and pain threshold will be changed abnormally after injected morphine over and over again. It suggested that the effect of morphine can not increased pain threshold after 7 days. But both of the treatment of electro-acupuncture and catgut embedding could return to normal, delay the developmental time of morphine-tolerance.2. The mice of morphine-tolerance have significant allorhythmia and negative activity. both of the treatment of electro-acupuncture and catgut embedding can change abnormal phenomenon.3. It is showed that the expression levels of Per1 and Per2 in SCN will be down-regulated after injected morphine. the treatment of electro-acupuncture can relieve this trend but the catgut embedding can not. |
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