Abstract:

Objective To screen the differentially expressed genes in type 2 diabetes mellitus model rats by transcriptomics sequencing， and to preliminarily explore the relationship between the differentially expressed genes and the syndrome manifestations of deficiency of both Qi and Yin. Methods Eighteen male SD rats were randomly selected as control group，and the other rats were fed with high-fat and high-sugar diet and injected with streptozotocin （STZ） in the abdomen to establish the type 2 diabetes models. On the 6th week， 31 successfully modeling rats were divided into model group （n=14） and drug evidence group （n=17） according to the principle of blood glucose and body mass balance. The rats in model group were fed with high sugar and high fat diet sequentially.The rats in drug evidence group were given Xiaokeling 2.2 g·kg^－1·d^－1 by gavage；the experiment last for 21 weeks.The body masses， food intakes， water intakes， mental state scores， sport scores， foot temperatures， grasping force， pulse amplitudes， respiratory rates， and tongue image integral of the rats were detected and the physiological signs of the rats during model preparation were evaluated.The fasting blood glucose（FBG） levels of the rats in various groups were detected by blood glucose meter. After the last administration， the rats were anesthetized， the blood samples were collected and the serum samples were separated.The levels of triglyceride （TG） and total cholesterol （TC） of the rats were detected by oxidase method.The levels of high density lipoprotein cholesterol （HDL-c） and low density lipoprotein cholesterol（LDL-c）of the rats were detected by direct method，and the levels of CD4， CD8，cyclic adenosine monophosphate （cAMP）， and cyclic guanosine monophosphate （cGMP） of the rats were detected by enzyme-linked immunosorbent assay （ELISA） method.The liver tissue of the rats was obtained，and the differentially expressed genes between model group and control group and between model group and drug evidence group were screened by transcriptomic sequencing. Results Compared with blank group， the physiological signs of the rats in model group were basically consistent with the syndrome manifestations of Qi and Yin deficiency； compared with model group， the some physiological signs of Qi and Yin deficiency of the rats in drug evidence group were significantly improved. Compared with blank group， the FBG，serum TC， TG and LDL-c levels of the rats in model group were significantly increased （P<0.01）， and the serum HDL-c level had no significant difference（P>0.05）；compared with model group， there were no significant differences in the above indexes of the rats in drug evidence group （P>0.05）.Compared with blank group， the level of serum CD4 and the CD4/CD8 ratio of the rats in model group were significantly decreased （P<0.01）， while the level of serum CD8 had no significant difference（P>0.05）；the cAMP level in serum and the ratio of cAMP/cGMP were obviously increased （P<0.01）， while the cGMP level in serum was significantly decreased （P<0.01）；compared with model group， the level of serum CD4 and the CD4/CD8 ^{ratio of the rats in drug evidence group were remarkably increased （P<0.01）， the cGMP level in serum was increased（P<0.01） and the cAMP/cGMP ratio was significantly decreased（P<0.01）， the cAMP level in serum had no significant difference （P>0.05）. Compared with blank group， the up-regulated expressed genes of the rats in model group mainly included early growth response factor 2 （Egr2），insulin-like growth factor binding protein 1 （Igfbp1） and a disintegrin and metallopeptidase with thrombospondin motifs 4 （Adamts4）； the down-regulated expressed genes mainly included G₀/G₁ switch 2 （G0s2）， midline 1 interaction protein 1 （Mid1ip1）， and basic helix-loop-helix family e40 （Bhlhe40）；compared with model group， the levels of the above differentially expressed genes in drug evidence group showed the opposite trend. Conclusion The syndrome of type 2 diabetes mellitus combined with deficiency of Qi and Yin is associated with up-regulation of Egr2， Igfbp1 and Adamts4 and down-regulation of G0s2， Mid1ip1，and Bhlhe40.}

Key words: Type 2 diabetes mellitus, Deficiency of Qi and Yin, Syndrome manifestation, Differentially expressed gene