| Objective:To investigate the effect of moxa-stick moxibustion on the expression of mineralocorticoid receptor,MR and glucocorticoid receptor,GR gene and protein in hippocampus and amygdala of rats with corticosterone deficiency of kidney yang.To reveal the possible molecular mechanism of moxa-stick moxibustion in improving kidney-yang deficiency syndrome.Methods:70 male Sprague-Dawley rats,weighing 200 ± 25 g,were divided into two groups: blank group(n = 20)and model group(n = 50),provided by the Animal experiment Center of Jiangxi University of traditional Chinese Medicine.Rats in the model group were given hydrocortisone(0.03mg)dose per gram of body weight once a day for 14 days to establish a kidney-yang deficiency model in the hindlimb(left and right hind extremities alternately injected with hydrocortisone).The model of kidney-yang deficiency was established in rats in the model group.When the model was completed,the rats who did not meet the requirements and died were eliminated according to the behavioral score,and the 40 rats who were successful in modeling were also divided into 2 groups according to the random numerical table.kidney yang deficiency model group(20 rats),moxibustion treatment group(20 rats).Moxibustion treatment group began to intervene moxibustion treatment from the 15 th day of the actual test.Before treatment,the hair at the acupoint was cut off in advance,and the moxa stick(8mm in diameter,long 12mm)made by the affiliated Hospital of Jiangxi University of traditional Chinese Medicine was suspended on the points of Guanyuan and Shenshu,and moxibustion with 2min preheated to preheat the local skin,then fixed at the above-mentioned points for mild moxibustion.The implementer will put the index finger around the acupoint,with a warm sense of heat is appropriate.Moxa-stick moxibustion for 20 minutes,once a day,7 times as a course of treatment,the interval of treatment rest for 2 days,and then the second course of treatment,a total of 14 treatment number of times.The other two groups of rats received the same binding fixation as moxibustion group rats.After two courses of treatment,draw materials The contents of ACTH,CRH,CORT in serum and MR and GR mRNA and protein expression in hippocampus and amygdala of rats in each group were detected by ELISA method and RT-PCR,Western blot technique respectively.Results:1.Comparison of body weight:(1)there was no significant difference in body weight of each group before modeling(P>0.05);(2)14 days after modeling,compared with model control group,there was no significant difference in body weight of rats in blank control group(P>0.05).There was a significant difference between the two groups(P< 0.01).2.Serum indexes:(1)Compared with the blank group,the expression of serum ACTH,CRH,CORT in the model group was lower than that in the blank group,and there was significant difference between the model group and the blank group(P < 0.01).(2)Compared with the model group,the expression of serum CRH,CORT in the moxibustion treatment group were significantly higher than those in the model group(P<0.01),and there was also a significant difference in the expression of serum ACTH in the moxibustion treatment group(P<0.05).(3)Compared with the blank group,there was no significant difference in the above indexes between the moxibustion treatment group and the blank group(P>0.05).3.mRNA expression:(1)Hippocampus:(1)Compared with the blank group,there was no significant difference in the expression of MR mRNA in the hippocampus of the model group,but the expression of GR mRNA in the model group was significantly lower than that in the control group(P<0.05).(2)Compared with the model group,the expression of MR,GRmRNA in hippocampus of moxibustion treatment group increased to some extent,but the difference was not statistically significant(P > 0.05).(2)Amygdala1 :(1)Compared with the blank group,the expression of MR,GR mRNA in the amygdala nucleus of the model group decreased significantly(P<0.01).(2)Compared with the model group,the expression of MR mRNA in amygdala of moxibustion treatment group increased significantly(P<0.01),and the expression of GR mRNA was significantly higher than that of model control group(P<0.05).(3)Compared with the blank control group,there was no significant difference in the expression of MR,GR mRNA in the amygdaloid nucleus between the moxibustion treatment group and the blank group(P>0.05).4.Protein expression:(1)Hippocampus:(1)Compared with the blank group,there was no significant difference in the expression of MR protein in hippocampus of each group(P>0.05),but the expression of GR protein in model control group was significantly lower than that in the control group(P<0.05).There was no significant difference in the expression of GR protein in hippocampus of rats treated with moxibustion(P>0.05)(2)Compared with the model group,the expression of GR protein in hippocampus of moxibustion treatment group was up-regulated and the difference was statistically significant(P<0.05).(2)Amygdala :(1)Compared with the blank group,the protein expression of MR,GR in the amygdala of the model group was significantly lower than that of the blank group(P <0.05).There was no significant difference in the expression of MR,GR protein in amygdala of moxibustion group to normal level(P>0.05).(2)Compared with the model group,the expression of MR,GR protein in amygdala in moxibustion treatment group was significantly higher than that in model group(P<0.05).Conclusion:1.In this experiment,after consulting the literature of various methods of modeling kidney-yang deficiency rats,the model of corticosterone kidney-yang deficiency rats was successfully established by intramuscular injection of hydrocortisone,which laid a good foundation for the development of this experiment.2.Moxibustion at Shenshu point and Guanyuan point can effectively improve the symptoms of yang deficiency in rats with deficiency of kidney yang.Under the intervention of warm and heat stimulation of moxibustion and specific acupoint effect,the hormone level of ACTH,CRH,CORT on the HPA axis increases,3.Moxibustion can regulating function of HPA axis gradually becomes normal,which makes the GR mRNA and protein expression in hippocampus and amygdala upregulat,also help the amygdala nucleus up-regulate the expression of MR mRNA and protein,thus participate in the negative feedback regulation of HPA axis,which may be one of the molecular mechanisms of moxibustion in improving kidney-yang deficiency syndrome. |
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