| Objective: To observe the effect of Wenyang Zhuyu Decoction on the expression of micro RNA-210 and its target gene Ephrin-A3 after cerebral ischemic injury in rats with kidney-yang deficiency syndrome,and to explore the prevention and treatment of Wenyang Zhuyutang’s brain deficiency in rats with kidney-yang deficiency syndrome Possible mechanism of blood damage.Methods: 192 adult healthy male SD rats were divided into a blank group(n=48)and a kidney-yang deficiency group(n=144)according to a random number table.After 21 days of intramuscular injection of hydrocortisone in the kidney-yang deficiency syndrome group,the plasma cyclic adenosine monophosphate(cAMP)and cyclic guanosine monophosphate(cGMP)levels were detected by enzyme-linked immunoassay(ELISA),and then the kidney-yang deficiency syndrome group Divided into sham operation group,model group and Wenyang Zhuyu decoction group(n=48),and divided into 4subgroups(n=12)according to the four sampling time points of 1d,3d,7d and14 d after operation.Except for the blank group,each group used the modified Longa suture method to prepare rat middle artery infarction models.The sham operation group was inserted into the internal carotid artery of the rat with a suture 8mm,and the blank group was fed with free diet;sham operation group and model group Distilled water was given to the stomach after the operation;the Wenyang Zhuyu Decoction group was given to the stomach after the operation.According to Longa’s 5-point system,rats were scored for neurological damage,and HE staining was used to assess the pathological damage of brain tissue;real-time PCR(Real-Time PCR)method was used to detect miR-210 and miR-210 in the ischemic side cortex of rats.The expression level of Ephrin-A3 mRNA;Western blot was used to detect the expression level of Ephrin-A3 protein in the cortex of the ischemic side of the rat.Use SPSS24.0 statistical software for data analysis.Results:(1)Specific indicators of kidney-yang deficiency: Compared with the blank group,the plasma cAMP content of rats in the kidney-yang deficiency group decreased and the cGMP content increased(P<0.01);compared with the model group,the yang was warmed after intragastric administration The content of cAMP in Zhuyu Decoction group increased,and the content of cGMP decreased(P<0.01).(2)Neurological deficit score: at the same time point,compared with the sham operation group,the model group and the Wenyang Zhuyu Decoction group had different degrees of neurological deficit(P<0.01);compared with the model group,Wenyang Zhuyu Decoction The neurological deficits in the group were improved to varying degrees(P<0.01).(3)Brain tissue morphological changes: the blank group and the sham operation group have complete neuron morphology without obvious pathological changes;the model group has atrophy and deformation of neurons,most of the neurons are necrotic and apoptosis,accompanied by inflammatory cell infiltration,showing obvious Ischemic damage changes;although there is still a small amount of neuron necrosis in the ischemic area of the Wenyang Zhuyu Decoction group,the cells are gradually densely arranged compared with the model group,the nucleus pyknosis and deep staining range is reduced,the nucleus is gradually clear,Improved sex cell infiltration.(4)The expression of miR-210 and Ephrin-A3 mRNA at the same time point: Compared with the sham operation group,the expression levels of miR-210 and Ephrin-A3 mRNA in the model group and the Wenyang Zhuyu Decoction group were significantly increased(P<0.01);and Compared with the model group,the expression of miR-210 in the Wenyang Zhuyu decoction group was higher than that of the model group,and the expression level of Ephrin-A3 mRNA in the Wenyang Zhuyu decoction group was lower than that of the model group,and the differences were statistically significant(P<0.01).(5)The expression of Ephrin-A3 protein at the same time point:Compared with the sham operation group,the expression of Ephrin-A3 protein in the model group and the Wenyang Zhuyu decoction group increased(P<0.01);compared with the model group,Wenyang The expression of Ephrin-A3 in the Zhuyu Decoction group decreased(P<0.01).Conclusion: Wenyang Zhuyu Decoction can improve the degree of kidney-yang deficiency and neurological deficit in rats with kidney-yang deficiency and cerebral ischemia,and by up-regulating the expression of miR-210 after cerebral ischemia injury,it can negatively regulate its target genes.It may be one of the mechanisms that induce angiogenesis after cerebral ischemic injury. |
