To Explore The Protective Effect Of Shennao Fuyuan Decoction On Hypoglycemia And Hypoxia PC12 Cells Based On The Mechanism Of Cell Pyrolysis

Objective:To observe the protective effect of Shennaofuyuan Decoction（SNFYD）containing serum on PC12 cells of Oxygen-Glucose Deprivation（OGD）and its effect on the expression of cytokines related to the Pyroptosis pathway of NLRP3/Caspase-1,IL-1beta/IL18,and to explore the effect related to Pyroptosis pathway death,cell culture,morphological examination and staining,biochemical detection and Western Blot were used in this study.And discuss its possible mechanism of impede pyroptosis death and mitigation of injury to protect PC12 cells,to provides experimental basis for its application in the treatment of ischemic stroke.Methods:1,Preparation of serum containing Shennaofuyuan Decoction:SD rats were fed Shennaofuyuan Decoction for 3 consecutive days,then collect abdominal aorta blood,serum was separated by centrifugation and frozen after inactivation.2,The culture and model establishment of PC12:PC12 cell line was purchased and cultured in DMEM-HG complete culture medium containing 10%FBS and1%bi-antibiotic.According to the detection index and experimental purpose,PC12 cell line was inoculated into the corresponding orifice plate.When the cells adhere to the seed plate and grow logarithmically,the sugar-free medium is replaced by complete culture medium and placed in a three-gas incubator at37℃.The culture is carried out with 95%N₂-5%CO₂.OGD lasted for 4 hours,then changed back to the original culture conditions,restored the supply of sugar and oxygen,and treated according to the grouping.3,Intervened by SNFYD containing serum culture medium:cells after OGD were divided into three groups,SNFYD group as experimental group,using complete culture medium containing 10%SNFYD containing serum for culture;negative control group was blank serum group,using 10%blank rat serum;positive control group chose NLRP3 inhibitor INF39 as intervention drug,adding as 1μM concentration to complete medium for culture.4,Relevant data:Cells in each group were observed under inverted microscope,apoptotic degree was detected under fluorescence microscope after AO/EB staining,activity was detected by CCK8 method,and expression of cytokines NLRP3,Caspase-1,and IL-1beta/IL18 in pyroptosis death pathway was detected by Western blot.Results:After OGD,PC12 was damaged obviously,and the cytokines of the pyroptosis pathway were increased obviously.The morphology and activity of PC12 cells were improved after intervention with SNFYD containing serum and INF39.Accordingly,the activation of NLRP3/Caspase-1 pyroptosis pathway was weakened,which indicated that the inhibition of this pathway was related to the improvement of cell status.Both SNFYD and INF39 could inhibit this pathway’s activation.It is suggested that Shennaofuyuan Decoction may protect nerve tissue by inhibiting NLRP3/Caspase-1 and downstream pyroptosis pathway and reducing inflammatory apoptosis of nerve cells,so as to achieve the goal of treating cerebral infarction.Conclusions:The protective effect of Shennaofuyuan Decoction on nerve tissue may be related to the inhibition of NLRP3/Caspase-1 and downstream cellular pathway.