| Purpose: The main research was on the effects of Powered acupuncture at "Gan Shu" on the behavior of circadian rhythm disorder C57BL/6J mice and the expression of circadian clock genes in the kidney.By acupuncture the "Gan Shu" point,observe the changes of the day and night behavior of the mice,and detect the expression of specific rhythm genes in the kidney to determine the effect of acupuncture at the "Gan Shu" point on the expression of specific rhythm genes in the peripheral circadian rhythm,and then from the gene To explore the mechanism of acupuncture at "Gan Shu" on C57BL/6J mice with circadian rhythm disorder.Methods: 72 C57BL/6J mice were randomly divided into four groups,which were divided into blank group,model group,acupuncture non-acupoint group and acupuncture liver group,with 18 rats in each group.After 7 days of acclimatization in light synchronization(L: D=12;12,Light: 08:00-20:00 Dark:20:08:00),the model group,acupuncture non-acupoint group and acupuncture liver group The mice were subjected to a short-time deprivation sleep method(L:D = 2: 2),modeled for 5 days,modeled successfully,and a sleep-wake cycle disorder model was obtained,after which the light synchronization was restored(L: D = 12;12).The blank group uses the light synchronization method throughout.After the successful modeling,on the 13 th day of the experiment,the blank group and the model group was only bundled for 20 minutes at 15:00every day,and the other two groups were subjected to acupuncture for 20 minutes at 15:00 every day for 5 consecutive days.The acupuncture group performed acupuncture at the bilateral "Gan Shu" points,and the non-acupoint group selected by the non-acupoint group was used for acupuncture at the non-acupoints fixed in the abdomen of the abdomen.The acupuncture method was inserted obliquely inward and downward into the 0.5cm,two Only the needle handle is connected to the output wire of the KWD-808 I pulse electro-acupuncture instrument,and the parameters of the electro-acupuncture instrument are adjusted to: dense wave 4/20Hz;the sparseness period is 6seconds,and the muscles of the acupuncture site or the lower limbs can be slightly twitched.After 11 hours of the last acupuncture,starting at 02:00 on the18 th day,the four groups were carried out at the same time.Three kidneys were taken from each group,and every 4 hours,a batch was executed until 22:00 on the 18 th day.The last batch was executed,and the 360-intelligent infrared camera was used to record and observe in real time.The spontaneous standing and activity times of the mice during the experiment were recorded.The m RNA levels of Cry1 and Cry2 genes in the kidney were measured by Real-Time-PCR method and the protein expression levels of Cry1 and Cry2 were determined by Western Blot.Cosine analysis and statistical analysis were used to verify whether gene expression was rhythmic.Results:1?Under the remote observation record of 360 cameras,the model group and the non-acupoint group mice showed arrogance,susceptibility,irregular stool,thin skin and dull fur,compared with the initial acclimation period.The number of hair and climbing has increased significantly,and it is obvious that there is a clear sound in the cage.The acupuncture group has recovered the characteristics of the nocturnal night after five days of acupuncture at the liver acupoint.The temperament is milder.The behaviors such as standing and mane are similar to those of the blank group.The fur is soft and the stool is normal.Using statistical analysis,after treatment,the number of spontaneous activities and the number of standing in the mouse within 5 min,the acupuncture group compared with the blank group,the non-acupoint group compared with the model group,the results were not statistically significant(P >0.05).Compared with the model group,the acupuncture group and the non-acupoint group,the number of activities and standing was significantly reduced,which was statistically significant(P<0.05).2?Under the Real-Time-PCR method,the expression of Cry1 and Cry2 genes in the blank group and the acupuncture group showed rhythm(P<0.3),and the expression of Cry1 and Cry2 genes in the model group and non-acupoint group showed rhythm.Sexual disorder(P>0.3).3 ? Western Blot protein detection showed that the difference between the model group and the blank group was statistically significant(P<0.05),indicating that the gene expression of the model group was not rhythmic.The difference between the acupuncture group and the model group was statistically significant(P<0.05),indicating that the protein expression of the acupuncture group was rhythmic,and the difference between the non-acupoint group and the acupuncture group was statistically significant(P < 0.05),showing that the non-acupoint group gene expression did not exhibit rhythm.Conclusion:1?The behavioral aspects of normal C57BL/6J mice and the expression of Cry1 and Cry2 genes in the kidney are rhythmic.2?.Acupuncture at the liver and acupoints helps the mice to return to normal behavior,while acupuncture does not have a role in non-acupoints.3?Acupuncture at "Gan Shu" points helped the rhythm genes Cry1 and Cry2 to return to normal in the kidney,which was rhythmic,and the disordered circadian rhythm cycle gradually returned to normal.The acupuncture does not reflect this effect.?... |
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