| Objective: The pathogenesis and prognosis of cerebral ischemia-reperfusion injury are closely related to the ups and downs of the spleen and stomach vigour.Medicine lotus leaf could regulate spleen and stomach,rise the effect of clear material,this research adopts the line switch method of animal model of cerebral ischemia reperfusion injury in rats main selection lotus leaf extract nuciferine base for treatment,explore the nuciferine base after the intervention,cerebral ischemia reperfusion injury in rats nerve function score,cerebral infarction,cerebral edema and ischemic brain tissue volume RhoA/ROCK ? signaling pathways,and serum inflammatory factor expression,for TCM clinical treatment of ischemic stroke in the lotus leaf provide experimental basis for further.Methods:1.SD rats were randomly divided into sham operation group(sham),model group(I/R),Fasudil treatment group(fasudil),and high,medium and low dose nuciferine treatment group(Nuciferine-H,Nuciferine-M,nuciferine-L)and received preadministration.The sham group and the model group were given purified water by gavage.The positive group received fasudil injection intraperitoneally.The high,medium and low dose nuciferine groups were given different doses of nuciferine gavage.2.Establishment of a rat model of focal cerebral ischemia(MCAO).The rat model of cerebral ischemia-reperfusion(MCAO)was established by the line bolt method after the last administration for 1 h.The neurological function score was measured 24 h later,the volume of cerebral infarction was detected by TTC staining,and the cerebral water content of rats was measured by dry and wet weighing method.The effects of nuciferine preadministration on cerebral ischemia-reperfusion injury were evaluated.3.By immune protein imprinting method and immunofluorescence method,to observe on the nuciferine dosage on cerebral ischemia reperfusion injury in rats cortex RhoA/Rock ? signal path expression.4.The expression levels of IL-4,IL-10,TNF-? and IL-1 in peripheral blood of rats with cerebral ischemia-reperfusion injury were detected by ELISA.Results: 1.After the preliminary experiment,I have mastered the production of the model of middle cerebral artery embolization cerebral infarction in rats by wire embolization.Neurological function score was performed 24 h after the successful cerebral ischemia modeling.Compared with the sham group,the neurological function score of the model group was significantly improved(P < 0.05).Compared with the model group,the scores of neurological impairment were significantly reduced in the high,medium and low dose nuciferine group and the fasudil group(P < 0.05).Compared with the fasudil treatment group,the high dose nuciferine treatment group could significantly reduce the neurological function defect score(P < 0.05).2.The cerebral edema was detected 24 hours after successful cerebral ischemia modeling.Compared with the sham operation group,the cerebral water content of the model group was significantly increased(P < 0.05).Compared with the model group,the cerebral water content in the high,medium and low dose nuciferine treatment group and the fasudil treatment group could be significantly reduced(P < 0.05).Compared with the fasudil treatment group,the cerebral water content in the high-dose nuciferine treatment group was significantly reduced(P < 0.05).3.TTC staining was used to detect the volume of cerebral infarction in rats with cerebral ischemia/reperfusion injury.This method was intuitive,effective,and relatively easy to operate.TTC ischemia test was performed 24 hours after successful cerebral ischemia modeling.No cerebral infarction was observed in the sham group.Compared with sham group,cerebral infarction volume in model group was significantly increased(P < 0.05).Compared with the model group,the volume of cerebral infarction of rats in the high,medium and low dose nuciferine treatment group and the fasudil treatment group was significantly reduced(P < 0.05).Compared with the fasudil treatment group,the cerebral infarction volume of rats in the high dose nuciferine treatment group was significantly reduced(P < 0.05).4.Western blotting method to detect each side cerebral ischemia of rats cortex section RhoA and Rock ? protein expression levels.Compared with the control group,the model group RhoA and Rock ? protein expression was significantly increased(P < 0.05).Compared with the model group,the fasudil treatment group and the nuciferine by high,medium and low dose nuciferine treatment group of RhoA and Rock ? protein expression level decreased obviously(P < 0.05).Compared with the fasudil treatment group,the high-dose nuciferine treatment group rats ischemic lateral cortex section RhoA and Rock ? protein expression significantly lower(P < 0.05).5.Immunofluorescence test between groups of ischemia rats lateral cortex section RhoA and Rock ? protein expression.Compared with control group,the model group and lateral cortex section RhoA Rock ? expression levels(P < 0.05);Compared with model group,the fasudil treatment group and nuciferine by high,medium and low dose treatment group of RhoA and Rock ? expression level decreased significantly(P < 0.05).Compared with the fasudil treatment group,the high-dose nuciferine treatment group rats ischemic lateral cortex section RhoA and Rock ? expression levels significantly lower(P < 0.05).6.The expressions of inflammatory cytokines TNF-?,IL-1,IL-4 and IL-10 in the serum of rats with cerebral ischemia-reperfusion injury were detected by ELISA,and the serum levels of IL-4,IL-10,TNF-? and IL-1 were significantly increased in the model group compared with the sham group(P < 0.05).Compared with the model group,serum IL-4,TNF-? and IL-1 levels in the fasudil treatment group and the nuciferine treatment group were significantly decreased and il-10 levels were significantly increased(P < 0.05).The serum levels of IL-4,TNF-? and IL-1 were significantly decreased and the serum levels of IL-10 were significantly increased(P < 0.05).Conclusion:1.The middle cerebral artery embolization model of rats was performed by thread embolization.The infarcted area was stable,the surgical effect was good,and the survival rate of rats was high.2.Lotus leaf extract nuciferine pretreatment with cerebral ischemia-reperfusion injury in rats can improve neurological function score,reduce cerebral infarction volume,improve cerebral edema in rats with cerebral infarction,and play a neuroprotective role.3.Lotus leaf extract nuciferine pretreatment of cerebral ischemia-reperfusion injury in rats,can cut RhoA/ROCK signaling pathways ? expression in ischemic brain tissue,improve neurologic recovery.4.Lotus leaf extract nuciferine preconditioning rats with cerebral ischemia/reperfusion injury can regulate the expression levels of IL-4,IL-10,TNF-? and IL-1 in peripheral blood,inhibit the inflammatory response after cerebral ischemia,and reduce the inflammatory damage. |
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