Analysis Of The Efficacy And Mechanism Of Ginsenoside Rg1 In Treating Acute Cerebral Stroke By Inhibiting CKLF1 From The Perspective Of The Theory Of "stasis And Toxins Causing Wind

Objective:We evaluated the pharmacological effects of ginsenoside Rg1 on acute cerebral ischemia-reperfusion injury and explored the protective effects and possible mechanisms of ginsenoside Rg1 on ischemia-reperfusion-induced blood-brain barrier injury to provide a theoretical basis for further development of ginsenoside Rg1 as a drug for the prevention and treatment of ischemic stroke.Methods:The SD rats were randomly divided into sham-operated group,model group,ginsenoside Rg1 low-dose group(10 mg·kg^-1),medium-dose group(20 mg·kg^-1),high-dose group(40 mg·kg^-1)and positive drug edaravone group(10 mg·kg^-1).A rat model of focal cerebral ischemia-reperfusion injury was constructed by thread embolism method,and the drug was administered by tail vein injection immediately after thread embolism was pulled out.Equal amounts of saline were given to the model and sham-operated groups.After 24 h of reperfusion,neurological deficits were scored by Zea Longa method,the area of cerebral infarction and the degree of edema in rats were determined by TTC staining,and neuronal death in the post-ischemic cortex was observed by Nissl staining.The expression of TNF-α,IL-1β,IL-6,CKLF1,ICAM-1 and VCAM-1 in the ischemic hemisphere was detected by ELISA,Western blot and immunofluorescence chemistry.The permeability of the blood-brain barrier was assessed by Evans blue staining,and the protective effect of ginsenoside Rg1 on the ultrastructure of the blood-brain barrier was verified by transmission electron microscopy.The expression of ZO-1,Occludin,MMP-2,MMP-9 and NF-κB p65/MAPK signaling pathway-related proteins in brain tissues of ischemic hemispheres was examined by Western blot and immunohistochemistry to investigate the mechanism of action of ginsenoside Rg1 in improving blood-brain barrier injury after cerebral ischemia-reperfusion.Results:Ginsenoside Rg1 intervention can significantly improve neurological deficit scores,reduce the rate of cerebral infarction and edema,reduce neuronal death,inhibit the protein expression of CKLF1,significantly reduce the expression of TNF-α,IL-1β,IL-6,ICAM-1,and VCAM-I,reduce the content of Evans blue in the brain on the ischemic side,effectively improve the blood-brain barrier permeability and attenuate the ultrastructural damage of the blood-brain barrier,and significantly down-regulated the expression of MMP-2,MMP-9,p-NF-κB p65 and MAPK signaling pathway-related proteins,and up-regulated the expression of tight junction protein ZO-1 and Occludin.Conclusion:CKLF1 is involved in the anti-stroke effect of ginsenoside Rg1.Ginsenoside Rg1ameliorates neurological impairment after cerebral ischemia-reperfusion,reduces ischemic inflammatory injury,and plays a protective role in maintaining the integrity of the blood-brain barrier.The regulatory mechanism may be related to the ability of ginsenoside Rg1 to inhibit the expression of CKLFl,suppress the activation of NF-κB p65 and MAPK signaling pathways,reduce the production of inflammatory factors and MMP,and upregulate the expression of tight junction-related proteins thereby reversing the disruption of the blood-brain barrier.