SENP1 In Pcricvte Protects Brain In Local Ischemia

Objective:The Senpl knockout in pericyte mice(Cspg4-Cre;Senplf/f)was constructed by Cre-loxp system and the rose bengal local ischemia model was used to exploring the pericytes Senpl function in cerebral ischemia.This study would provide an important theoretical basis for stroke.Method:(1)Cspg4-Cre;Senplf/f mice were obtained by mating Cspg4-Cre mice with Senplf/f mice.The density of neurons,Y-Maze tests and rotarod test were used to explore the effect of pericytes knocking out Senpl on motor performance,learning and memory.(2)The model of local cerebral ischemia in mice was made by Rose Bengal.The infarct volume was quantified by TTC staining;The density of neurons was counted in penumbra field and ischemic core by immunofluorescence staining after 24 hours.(3)After 24 hours of local cerebral ischemia,the mice were subjected to a rotating test and Y-Maze behavioral test to investigate the effect of pericytes knockout of Senp1 on exercise capacity,learning and memory in mice.(4)3D reconstruction of cerebral blood vessels by two-photon in vivo imaging technique was used to detecting vessel density and two-photon line scanning technique was used to measuring microvascular blood flow velocity,diameter and perfusion volume of the cerebral cortex.These methods were used to explore the basic characteristics of cerebral microvessels after pericytes knockout Senp1 under normal conditions.The two-photon in vivo time-lapse imaging technique was used to detecting the degree of vascular embolization in penumbra at 3h,6h,and 24h after Rose Bengal-induced cerebral ischemia.(5)Western blot was used to detecting the expression of Spectrin and Calcineurin proteolytic fragments after 24 hours of cerebral ischemia.Explore effects of the pericytes knockout of Senpl on the degree of functional impairment of neurovascular unit.The morphology of pericytes in the cerebral ischemic penumbra was detected by immunofluorescence technique to investigate the effect of pericytes knocking out Senpl on pericytes morphology.Result:(1)Immunofluorescence results showed that mice with knockout of Senpl in pericytes were successfully obtained by mating of Cspg4-Cre mice and Senp1f/f mice.Under normal conditions,the knockdown of Senpl in pericytes has no effect on the motor ability and learning and memory function of mice.(2)The volume of cerebral ischemic infarction increased significantly after pericytes knockout of Senpl in the model of cerebral local ischemia.The density of neurons in penumbra field and ischemic core is significantly reduced after the knockout of Senpl in pericytes.(3)After 24 hours of regional cerebral ischemia,the motor ability of the knockout Senpl mice after ischemia was decreased,but had no effect on learning and memory.(4)there was no change in microvessels density after pericytes of Senp1 under normal conditions.The results of two-photon line scan showed that the diameter,flow rate and perfusion volume of brain micro vessels did not change under normal conditions.In Rose Bengal cerebral ischemia model,two-photon scanning showed that the cerebral vascular occlusion was severe after the pericytes knocked out Senp1,and the severity increased with time.(5)The expression of Spectrin and Calcineurin proteolytic fragments significantly aggravated after the knockout of SENP1 in local ischemia.Endfoot of pericytes knocking out Senpl was disrupted after ischemia.Conclusion:By construction of pericytes-Senp1 knocking out transgenic mice,we determinate of neuro-vascular unit function and neurological indicators in the rose bengal local ischemia model,indicates that pericytes-SENP1 plays an important protective role in cerebral ischemia.