| Objective:This study utilizes a model rat model of cerebral ischemia-reperfusion injury(MCAO),using the PERK/Nrf2 signaling pathway as the research target,to explore the mechanism of action of lotus leaf alkaloid in the intervention process of model rats,and provide more sufficient experimental basis.Methods:1.30 male SPF grade SD rats were used in the experiment.Divided into sham surgery group(Sham group),model group(MCAO group),and model+lotus leaf alkaloid group(MCAO+NF group).Pre administration: Sham group and MCAO group were given 2ml of physiological saline by gavage each time;The MCAO+NF group was given 2ml of lotus leaf alkaloid solution by gavage once a day for one week.2.Establish a MCAO rat model using the suture method.Conduct NIHSS assessment on each group of rats;Dry wet weighing method was used to measure the brain water content of rats in each group;Conduct TTC staining experiments to calculate the proportion of cerebral infarction in each group of rats.Based on the above aspects,evaluate the impact of lotus leaf alkaloid intervention on MCAO model rats.3.Detect the expression levels of PERK and Nrf2 genes in each group of rats through PCR reaction;Using ELISA method to detect the expression levels of oxidative stress related factors SOD and MDA;Using immunofluorescence experiments to investigate the expression of lotus leaf alkaloids on the quantity and localization of PERK/Nrf2 protein.Results:1.The model of cerebral ischemia was successfully established using thread occlusion method.The NIHSS experiment results showed that the Sham group rats exhibited normal performance and no neurological deficits 24 hours after the establishment of the rat model;Compared with the Sham group,the NIHSS score of the MCAO group showed severe neurological damage in rats,with a statistically significant difference(P<0.05).Compared with the MCAO group,the NIHSS score of the MCAO+NF group showed a significant decrease in neurological deficits in rats(P<0.05).2.Brain water content detection results: Compared with the Sham group,the proportion of brain water content in MCAO group rats significantly increased(P<0.05).Compared with the MCAO group,the proportion of brain water content in the MCAO+NF group significantly decreased after lotus leaf alkaloid intervention(P<0.05).3.TTC staining results: The brain slices of Sham group rats showed normal red color and no signs of cerebral infarction;The majority of brain slices stained in MCAO group were gray white,and the proportion of cerebral infarction was significantly increased compared to Sham group(P<0.05);Compared with the MCAO group,the gray white area of the brain slices in the MCAO+NF group showed a significant reduction in infarct size(P<0.05).4.PCR reaction detection of PERK/Nrf2 gene expression levels: Compared with the Sham group,the expression levels of PERK and Nrf2 genes in the reaction system of MCAO group rats increased(P<0.05);Compared with the MCAO group,the expression levels of PERK and Nrf2 in the reaction system of the MCAO+NF group model rats decreased(P<0.05).5.ELISA method for detecting the SOD activity and MDA content of oxidative stress factors: Compared with the Sham group,the SOD activity in the serum of the MCAO group significantly decreased while the MDA content significantly increased(P<0.05);Compared with the MCAO group,the SOD activity was significantly increased and the MDA content was significantly decreased in the MCAO+NF group(P<0.05).6.Immunofluorescence detection of PERK/Nrf2 protein expression and localization results: Compared with the Sham group,the expression levels of PERK and Nrf2 in the MCAO group rats’ test samples increased(P<0.05);Compared with the MCAO group,the expression levels of PERK and Nrf2 in the detection samples of the MCAO+NF group model rats decreased(P<0.05).Conclusion:1.After the intervention of lotus leaf alkaloids in rats with cerebral ischemia-reperfusion injury,the NIHSS score of the rats was improved,and the brain water content was significantly reduced.At the same time,the experimental part measured a decrease in the area of cerebral infarction,which has a neuroprotective effect.2.After the intervention of lotus leaf alkaloids in rats with cerebral ischemia-reperfusion injury,it can reduce the generation of MDA,increase the activity of SOD in the rat brain,and inhibit the occurrence of oxidative stress.3.After the intervention of lotus leaf alkaloids in rats with cerebral ischemia-reperfusion injury,the expression of PERK and Nrf2 genes is promoted to achieve the goal of antioxidant stress,and the nuclear translocation of Nrf2 is promoted,reducing the degree of brain injury. |
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