| Objective:To explore the mechanism of temporal and dynamic variety of acupoint reactivity based on the change of TRPV1&TRPV4 receptors on different parts after acupuncture.Method:The experiment had two steps. Firstly, the clean-male SD rats were synchronized to normal light/dark cycle for 7days. As the result of basic pain threshold test, the rats were randomly allocated to model group and blank group. The rats in model group were injected complete Freund's adjuvant(CFA) on the right footpad and randomly allocated to model-control group and model-acupuncture group. According to different time point of acupuncture, each group was divided into 6 subgroups-ZT0(7:00),ZT4(11:00),ZT8(15:00),ZT12(19:00),ZT16(23:00), ZT20(3:00). At 6 above-mentioned time points, each rat in model-acupuncture group was acupunctured "Zusanli(ST36)" for 30 min, twisted 1 min every 5 min,120 per/min. The rats in blank group and model-control group were bound for 30 min without acupuncture. When acupuncture therapy was finished, the changes of pain threshold on the different time points were detected by heat tail flick test. Secondly, as the result of analysis of variance(ANOVA), the time points, which the peak and valley point was occurred on curve after acupuncture, were chosen to further experiment. The method of grouping and modeling was the same as the first step. The acupuncture therapy lasted for 7 days. When the 7th therapy was finished, the samples should be taken immediately. And then, the expresses of TRPV1&TRPV4 on partial acupoint, dorsal root ganglion and spinal cord were detected by immunohistochemistry. That was to explore whether the temporal and dynamic variety of acupoint reactivity was related to TRPV1&TRPV4.Results:(1)Temporal factor had a significant effect on the pain threshold of the blank group(F=9.205, P=0.00, P<0.05). The peak point of the pain threshold appeared at ZTO and the valley point of its appeared at ZT4 and ZT12.(2)The pain threshold of the rats in model-control group decreased obviously and its circadian rhythm became disorder. Temporal factor had no significant effect on the pain threshold of the model-control group (F=1.155,P=0.354>0.05).(3)The pain threshold of the rats in model-acupuncture group increased obviously after acupuncture therapy. Temporal factor had a significant effect on the pain threshold of the model-acupuncture group(F=30.414, P=0.000<0.05). The most obvious time point was at ZT8 and the less obvious time point was at ZT16.(4)On the partial skin of acupoint:Compared with the blank group, the content of TRPV1&TRPV4 in the model-control group increased obviously(P<0.05 or P<0.01) at ZT8. Compared with the model-control group, the content of TRPV1&TRPV4 in the model-acupuncture group decreased obviously(P<0.05) at ZT8. Compared with the blank group, the content of TRPV1&TRPV4 in the model-control group increased obviously(P<0.05) at ZT16. Compared with the model-control group, the content of TRPV1 in the model-acupuncture group decreased obviously(P<0.05) at ZT16. Whereas the content of TRPV4 was not significant difference between the model-control group and model-acupuncture group at ZT16(P>0.05).(5)On dorsal root ganglion:Compared with the blank group, the content of TRPV1&TRPV4 in the model-control group increased obviously (P<0.01) at ZT8. Compared with the model-control group, the content of TRPV1&TRPV4 in the model-acupuncture group decreased obviously(P<0.05 or P<0.01) at ZT8. Compared with the blank group, the content of TRPV1&TRPV4 in the model-control group increased obviously(P<0.05 or P<0.01) at ZT16. Compared with the model-control group, the content of TRPV1 in the model-acupuncture group decreased obviously(P<0.05) at ZT16. Whereas the content of TRPV4 was not significant difference between the model-control group and model-acupuncture group at ZT16(P>0.05).(6)0n spinal cord:Compared with the blank group, the content of TRPV1&TRPV4 in the model-control group increased obviously (P<0.05 or P<0.01) at ZT8. Compared with the model-control group, the content of TRPV1 in the model-acupuncture group decreased obviously(P<0.01) at ZT8. Whereas the content of TRPV4 was not significant difference between the blank group and model-control group at ZT8(P>0.05). The content of TRPV1&TRPV4 in the model-control group increased obviously(P<0.01) at ZT16. Compared with the model-control group, the content of TRPV1&TRPV4 in the model-acupuncture group decreased obviously(P<0.05) at ZT16.Conclusion(1) The of normal rats has shown rhythmical fluctuation. And temporal characteristic showed double valley structure and the valley point respectively appeared at ZT4 and ZT12.(2) Acupoint reactivity had temporal dynamic variety. Acupuncture could increase the pain threshold of the rat in all different time points, but its extent had an effect on temporal characteristic. It showed peak-valley structure. That was to say, acupoint reactivity had time difference. The best time of acupuncture was at ZT8 and the worst was at ZT16.(3) Temporal and dynamic variety of acupoint reactivity might be related to the kind and quantity of excitation of TPRV1&TPRV4 on the partial acupoint, dorsal root ganglion and spinal cord after acupuncture therapy. |
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