The Effect Of Stellate Ganglion Block On Expression Of Apoptosis-related Proteins And Inflammatory Cytokines In Rat Cardiopulmonary Bypass

Objective: To build SD rat right starlike ganglion block(SGB)model,SD rat heart beating cardiopulmonary bypass(CPB)model,SD rat right starlike ganglion block model combined with cardiac arrest cardiopulmonary bypass model(CPB+SGB).In order to detect the effects of right starlike ganglion block on the hippocampus of SD rats under cardiopulmonary bypass,the mechanisms of related inflammatory pathways and apoptotic pathways were investigated.Methods: Seriously select 24 adult male SD rats of SPF level,the body weight measurement is about 350 g.The 24 SD rats were randomly divided into 4groups.Including: sham operation group(group S),simple right stellate ganglion block model group(group SGB),pure heart beating cardiopulmonary bypass model group(group CPB),before operation,right starlike ganglion block combined with heart beating cardiopulmonary bypass model group(group CPB+SGB),four groups were 6.Shallow anesthesia,group CPB+SGB and group SGB were used to construct the right stellate ganglion block model,and then deepen anesthesia.Normal anesthesia was performed in group S and group CPB.After the above operations,the above four groups performed the punctureand concatenation of the right jugular vein and the inferior femoral artery in SD rats.The following steps were followed: group CPB and group CPB+SGB successfully performed cardiopulmonary bypass and stopped the experiment two hours later;and group S and group SGB only needed close observation and stopped the experiment after two hours.The respiratory frequency of the rats,skin color of the limbs,heart rate,pulse,blood pressure,pain,blood oxygen,pupil and corneal reflex were closely monitored and recorded during the experiment.At the end of the experiment,the hippocampal tissues of rats were rapidly separated,and some hippocampal tissues were fixed with paraffin embedded sections after fixed with neutral Formaldehyde Solution.After HE staining,the pathological changes of hippocampus in rats were examined.The expression of Bax and Bcl-2 protein in hippocampus of rats was detected by immunohistochemistry.In addition,some hippocampal tissues were stored at-80 ? for detection of Caspase-3 gene expression by reverse transcription polymerase chain reaction(RT-PCR).Another part of hippocampus was preserved at-80?,and was used to detect the expression of Caspase-3 protein by Western blot.Results:The rat model of simple right stellate ganglion block,rat heart beating cardiopulmonary bypass model,right starlike ganglion resistance combined with cardiac beating cardiopulmonary bypass model before operation were successfully constructed.HE results showed that in the S group and SGB group,the morphology of CA1 cells in the hippocampus was normal.There is no change in the basic structure of the neuron.Most of the cell groups are neatly arranged.The cell membrane and internal organelles are relatively intact.The nucleolus of the nucleus is clear and unchanged.There are many organelles around the nucleusand normal structure.he ultrastructure of hippocampal neurons in the CPB and CPB+SGB groups was obviously damaged,the cells of the hippocampus were swollen,the cells were arranged in disorder,the boundaries between the nuclei and the membrane were not clear,the nucleus nucleolus dissolve,the nucleus had nuclear condensation,and the organelles around the nucleus decreased obviously.Group CPB and group CPB+SGB showed obvious apoptosis in hippocampal neurons.Compared with group CPB,the degree of injury in group CPB+SGB was relatively reduced.The results of ELISA displayed that the thickness of TNF-alpha and IL-6in the adtevak of CPB and CPB+SGB groups rised significantly at T1 and T2 time point(P<0.05).Compared with the CPB group,the thickness of TNFalpha and IL-6 in the adtevak of CPB+SGB groups decreased significantly at T1 and T2 time point(P<0.05).The immunohistochemical results displayed that compared with the S group,the amount of Bax protein expression in group CPB and CPB+SGB rised distinctly(P<0.05).Compared with the CPB group,the amount of Bax protein expression in CPB+SGB group decreased distinctly(P<0.05),and the amount of Bcl-2 protein rised distinctly(P<0.05).The results of PCR displayed that compared with the S group,the amount of Caspase-3 gene expression in group CPB and CPB+SGB rised distinctly(P< 0.05).Compared with the CPB group,the amount of Caspase-3 gene expression in CPB+SGB group decreased distinctly(P<0.05).The results of WB displayed that compared with the S group,the amount of Caspase-3 protein expression in group CPB and CPB+SGB rised distinctly(P< 0.05).Compared with the CPB group,the amount of Caspase-3 protein expression in CPB+SGB group decreased distinctly(P<0.05).Conclusions:Cardiopulmonary bypass can lead to the apoptosis of hippocampus grow in number and cause brain damage in rats.Stellate ganglion block can abate the chroma of TNF-alpha and IL-6 in plasma,thereupon then accommodating the inflammatory response of the body.Through down regulating the expression level of Bax and up regulating the expression level of Bcl-2,the amount of Caspase-3 protein expression was reduced,and ultimately apoptosis was reduced.Finally,stellate ganglion block can a certain extent protect the brain tissue of cardiopulmonary bypass rats under beating heart condition.