Effect Of Shennao Fuyuan Decoction On HPA Axis Function Of MCAO Reperfusion Rats

Objective:To observe the effect of Shennao Fuyuan Decoction on the expression of HPA axis in rats with middle cerebral artery occlusion and reperfusion,and to study the biological basis of Shennao Fuyuan Decoction for kidney and brain protection,and to guide the treatment of nervous system diseases for the theory of "reinforcing kidney and brain" Provide experimental basis.Methods:150 rats with specific pathogen free grade SD rats were randomly divided into 30 groups,30 sham operation groups and 90 MCAO model preparation groups.The blank group and the sham operation group were divided into 3d,7d,14 d and 3 phase groups according to the time of death,with 10 in each group.In the model preparation group,the model of focal cerebral ischemia with middle cerebral artery occlusion was established by modified LongaEZ and other methods.The model was reperfused after ischemia for 2 hours.After 2 hours of waking,the neurological function score was 1 to 3 points.success.The sham operation group was treated as a simple operation without threading.After the model was completed,the rats after successful modeling were divided into two groups according to the random number table method,namely,the model group and the Shennao Fuyuan Decoction group,and then divided into three groups after ischemia and reperfusion according to the time of death.7d,14 d,3 time phase groups,each group guarantees 6 animals successfully in each phase.The drug intervention was started on the 2nd day after modeling,and the Shennao Fuyuan Decoction group was administered to Shennao Fuyuan Decoction.The amount of fluid was 10 ml/kg per time.The blank group,the model group,and the sham operation group were intragastrically administrated with an equal volume of distilled water,and each group was intragastrically administered twice a day until sacrifice.After the intervention,the serum CRH and CORT levels were deteand adrenal gland were observed by HE staining.The expression of GR in hippocampus was detected by immunohistochemistry.The expression of CRHmRNA in hypothalamic paraventricular nucleus was detected by qPCR.Results:(1)Neurological function score: The m-NSS score of the sham operation group was not significantly higher than that of the blank group(p>0.05),and the difference was not statistically significant.The m-NSS scores of the model group and the Chinese medicine group were significantly higher than those of the blank group(p < 0.05).Compared with the model group,the score of the Chinese medicine group was lower than that of the model group at the same time point(p<0.05 or p<0.01),and the difference was statistically significant.(2)HE staining was used to observe the pathological changes of brain and adrenal gland: Compared with the model group,the morphological destruction of the cerebral cortical neurons in the ischemic side of the Chinese medicine group was light,and the neurons showed little vacuole-like changes.The arrangement was still neat,the number of remaining neurons was large,and the surrounding interstitial lesions were light,cortical neurons and nerves.The distribution of glial cells is roughly normal;Compared with the model group,the structure of adrenal cortex in the Chinese medicine group was basically normal,the cell arrangement was neat,and there was a little vacuole-like change in the cells.The nucleus,cytoplasmic swelling and peripheral interstitial necrosis were lighter than the model group.(3)Immunohistochemical method was used to detect the expression of GR in hippocampus of rats: compared with the blank group,the expression of GR in hippocampus of sham operation group was not significantly decreased(p>0.05),the difference was not statistically significant;the expression of GR in hippocampus of model group and Chinese medicine group Below the blank group(p<0.05).Compared with the model group,the expression of GR in the hippocampus of the Chinese medicine group was higher than that of the model group at the same time point(p<0.05 or p<0.01),and the difference was statistically significant.(4)ELISA method was used to detect serum CORT and CRH levels in rats: compared with the blank group,the serum CORT and CRH levels in the sham operation group were not significantly increased(p>0.05),the difference was not statistically significant;the model group and Chinese medicine The levels of serum CORT and CRH in the group were significantly higher than those in the blank group(p<0.05).Compared with the model group,the serum levels of CORT and CRH in the Chinese medicine group were lower than those in the model group at the same time point(p<0.05 or p<0.01),and the difference was statistically significant.(5)The expression of CRH mRNA in the PVN region of rat hypothalamus was detected by qPCR: Compared with the blank group,the CRH mRNA level in the PVN region of the sham operation group was not significantly increased(p>0.05),the difference was not statistically significant;The CRH mRNA level in the PVN area of the Chinese medicine group was significantly higher than that in the blank group(p<0.05).Compared with the model group,the CRH mRNA in the PVN region of the Chinese medicine group was lower than that in the model group at the same time point(p<0.05 or p<0.01),and the difference was statistically significant.Conclusion:1.The cerebral ischemia-reperfusion injury in rats continued to activate the HPA axis,and the CORT level increased and was positively correlated with the pathological damage of brain and adrenal gland.2.Shennao Fuyuan Decoction can down-regulate the expression of CRH and CORT in rats after cerebral ischemia-reperfusion,and up-regulate the level of GR in hippocampus,which can effectively regulate the disorder of HPA axis,improve the symptoms of neurological deficit in rats with ischemia-reperfusion injury,and promote pathological repair.Played an effective therapeutic role.