Construction Of Animal Model Of Kidney-Yang Deficiency Constitution And Related Transcriptome Study

Objective:Using the method of congenital endowment and acquired environment,a rat model of Kidney-Yang Deficiency constitution was established,which conforms to the theory of etiology and pathogenesis of traditional Chinese medicine.The rationality and scientificity of the model were demonstrated by evaluating the level of hormones related to hypothalamus-pituitary-target gland axis in animals,and the expression level of genes related to Kidney-Yang Deficiency in the Kidney-Yang Deficiency rat model was observed to further evaluate the model,at the same time,a new molecular level of thinking was provided for the diagnosis and treatment of Kidney-Yang Deficiency.Methods:1.Establishment and evaluation of Kidney-Yang Deficiency rat model: 130 SPF-grade 22-month-old SD rats of female and 90 male were observed and screened for cold-heat tendency and general traits.The rats with the most Kidney-Yang Deficiency tendency(staying in the area of 40 C of cold-hot plate for 500 s-600 s and their behavioral manifestations conformed to the clinical manifestations of Kidney-Yang Deficiency)were mated and fed and fed with cold food for pregnant rats.The offspring were used as the model group,and the offspring were used as the normal control group,while the rats remained in the hot and cold plate for less than 300 seconds.Six Kidney-Yang Deficiency model rats and six normal rats were randomly selected after the birth of their offspring.Serum CORT,T3,LH and other hormones related to hypothalamus-pituitary-target axis were detected and evaluated.2.Combined with a large number of previous experimental studies and HowNet search,we screened ANXA1,C-fos,DAPK2,FHIT,IGF1,LAIR1,Lpl1,MBNL1,SOCS2.Six normal rats and six Kidney-Yang Deficiency model rats were randomly selected from the normal group and the model group after the successful establishment of the model.SYBR Green fluorescence quantitative PCR was used to detect and evaluate the Kidney-Yang Deficiency model rats from the level of molecular biology.Results:1.General character and weight: the mental state of rats was still good after modeling,the reaction was relatively slow,prefer static,the hair color was dark and disordered,and the weight of Kidney-Yang Deficiency constitution model rats was significantly higher than that of normal rats(p<0.01),the difference was significant.2.Detection of serum hormone level: Compared with the normal group,the hypothalamus-pituitary-target axis-related hormones in Kidney-Yang Deficiency model rats:(1)The level of Corticosterone decreased significantly,the difference was significant(p<0.05);(2)The level of LH was significantly decreased(p<0.05);(3)The level of T3 was significantly decreased,the difference was significantly(p<0.05);3.RT-PCR: compared with the normal group,the quantitative results of the gene of the Kidney-Yang Deficient physique rats showed that:(1)There was a significant difference between the level of LAIR1 and the difference of p = 0.028(p <0.05);(2)the level of Lpl1 was significantly higher,p=0.000(p <0.05);(3)The level of FHIT was significantly higher,the difference was significant,and the difference of p=0.011(p <0.05)was of statistical significance.Conclusions:1.The evaluation of a biological index of Kidney-Yang Deficiency model rats shows that the method of combining innate endowment with acquired environment in this experiment can establish the animal model of kidney-yang deficiency.2.In this study,nine genes related to Kidney-Yang Deficiency were screened out from the results of differential gene expression profiles of the population with Kidney-Yang Deficiency,and there were significant differences in the expression level of three genes among the nine genes of the Kidney-Yang Deficiency model rats,and there was a significant difference in the expression trend of three genes.These six genes are mainly involved in immune response,energy metabolism,tumorigenesis,aging and other physiological processes.It shows that the model is similar to human kidney-yang deficiency in molecular biology level,and further demonstrates the rationality of the model.