Immune Inflammatory Regulation Of Mechanical Thrombectomy Therapy For Large Vessel Occlusive Cerebral Infarction

Objective:Acute ischemic stroke(AIS)is a condition caused by impaired blood supply to the brain,leading to rapid loss of brain function due to neural damage.It triggers a robust inflammatory response and is associated with unfavorable patient outcomes.This study aims to analyze the neuroprotective effects of identifying endovascular intravascular mechanical thrombectomy surgical treatment for effectively restoring IS patients’ blood flow and the immunoregulatory targets with adjuvant therapeutic potential.This contributes to the development of future strategies for alleviating secondary injuries and promoting recovery in ischemic stroke through rehabilitation techniques.Methods:Gene expression data of AIS patients and healthy controls in GSE16561,GSE22255,GSE122709 and GSE55937 datasets were collected from the gene expression omnibus database.Differential expression analysis was performed to identify differentially expressed m RNAs(DEm Rs),lnc RNAs(DElnc Rs)and mi RNAs(DEmi Rs)between AIS and controls after normalizing the processed data.The DEm Rs from the three datasets were intersected to obtain the intersected DEm Rs,which were subjected to biological enrichment analysis.The lnc RNAs targeting regulatory DEmi Rs were predicted using the DIANA and NPInter databases,Targetscan database predicted m RNAs targeted to regulate Demi Rs,Immport database for immune related m RNAs.In turn,a lnc RNA /mi RNA / m RNA ce RNA network that is differentially expressed and immune related in AIS was constructed.The diagnostic predictive ability of target DEm Rs in the ce RNA network was evaluated by ROC curve and nomogram.Evaluation of immune cells in each sample was performed by calculating ss GSEA,and abnormal levels of immune cells in the AIS were subjected to correlation analysis with target DEm Rs.Then a cohort of 64 patients with AIS,recruiting 64 gender and age-matched healthy control individuals were collected from the same period,and conducting validation of key outcomes.A total of 428 patients with AIS who underwent endovascular intravascular mechanical thrombectomy intervention were included.Hematological tests and scale assessments were conducted on patients within 24 hours of admission or the following morning,as well as 3 days after treatment.Patients were categorized into mild(NIHSS score<4),moderate(score 4-15),and severe(score > 15)groups based on preoperative NIHSS scores.Additionally,based on m TICI grade(IIb-III)and m RS score,patients were divided into good prognosis(m RS0-2)and poor prognosis(m RS 3-5)groups.Clinical data were compared between groups,and immune cells and the PI3K-AKT signaling pathway were analyzed for AIS patients with good and poor prognosis based on post-treatment m RS scores.Eighteen male rats were randomly equally divided into sham operation group(control group): MCAO group(IS group): MCAO + intravascular mechanical thrombectomy treatment group(thrombectomy group).Rats were subjected to middle cerebral artery occlusion to construct the MCAO model,and after 15 min of middle cerebral artery occlusion in rats of the intravascular mechanical thrombectomy treatment group,the monofilament was withdrawn for intravascular mechanical thrombectomy treatment.The neurological deficits of rats were evaluated using the Longa score,and the infarct size in the white region of rat brain tissue was observed by TTC staining.Neutrophils and NK cells were analyzed using flow cytometry.The key target DEm Rs was quantified through quantitative real-time polymerase chain reaction(q RT-PCR).The PI3K-AKT signaling pathway was assessed using Western blotting.Inflammatory cytokine levels were measured using enzyme-linked immunosorbent assay(ELISA).Results:(1)A total of 4988 DEm Rs were identified in the GSE16561 dataset: 3853 DEm Rs were identified in the GSE22255 dataset: 7224 DEm Rs were identified in the GSE122709 dataset,thus 388 intersected DEm Rs were obtained.The 9 targeted DElnc Rs,19 DEmi Rs and 12 targeted DEm Rs were obtained by the targeting prediction results and constructed as a ce RNA network.FNAR1,PAK2,PIK3 CG,RAF1,and SYK were mainly involved in PI3K-AKT signaling pathway.The combined ROC curve and nomogram results suggested that FNAR1,PAK2,PIK3 CG,and SYK might be important target DEm Rs with higher diagnostic predictive roles in AIS patients.(2)The levels of various immune cells,such as mast cells,natural killer cells,and neutrophils,are higher in AIS than in healthy controls.The results of the correlation study revealed a stronger link between the major target DEm Rs and neutrophils and natural killer cells.There was a significant increase in natural killer cell and neutrophil levels in AIS patients.The PI3K-AKT signaling pathway exhibited substantial inhibition.(3)Patients were categorized based on preoperative NIHSS scores into mild group(NIHSS<4)with 17 cases,moderate group(NIHSS 4-15)with 276 cases,and severe group(NIHSS > 15)with 135 cases.There were significant differences(P < 0.05)among the three groups in terms of compensation for good collateral circulation,cerebral hemorrhage,OPT time,ASPECTS score,swallowing function assessment,and neutrophil/lymphocyte ratio.(4)Among patients with m TICI IIb-III,based on m RS scores,they were divided into a good prognosis group(n=120)and a poor prognosis group(n=116).In comparison to the group with a bad prognosis,the fraction of collateral circulation was much higher in the group with a favourable prognosis.The proportions of ASPECTS score,postoperative high-density sign,lung infection,bleeding problems,and neutrophil/lymphocyte ratio were significantly lower in the group with a good prognosis.Natural killer cells and neutrophils were significantly reduced in the good prognosis group(P<0.001),while p-PI3 K and p-AKT were significantly elevated in the good prognosis group(P<0.001).(5)Compared with healthy controls,the levels of natural killer cells and neutrophils in patients with AIS were significantly higher,and compared with before surgery for AIS,the levels of natural killer cells and neutrophils decreased after treatment,but remained higher than those in controls.Obvious infarct zone was seen in the brain tissue of IS rat model.Statistical analysis of the Longa score and TTC staining results revealed that,compared with IS rats,the rats in the thrombectomy group showed a significant decrease in neurological deficit scores and a significant reduction in infarct rates.(6)Assays results on day 12 after transient MCAO revealed that,compared with sham controls,the expression levels of PIK3 CG,PAK2,IFNAR,as well as PI3K-AKT signaling pathway in the rat brain tissues were all significantly decreased in IS:compared with IS,PIK3 CG,PAK2,IFNAR,as well as PI3K-AKT significantly increased in the rats of the thrombectomy group.SYK was increased in IS,and decreased after thrombectomy.Conclusions:(1)By constructing a ce RNA network of lnc RNA/mi RNA/m RNA,immune related PIK3 CG,PAK2,IFNAR1,and SYK were identified to have higher diagnostic predictive effects in IS patients,possibly affecting the PI3K-AKT signaling pathway.Natural killer cells and neutrophils are elevated in IS and have the strongest correlation with target DEm Rs.(2)Compensation for good collateral circulation,ASPECTS score,and neutrophils/lymphocytes may all be influencing factors for the good prognosis of AIS intravascular intravascular mechanical thrombectomy surgery.The reduction of natural killer cells and neutrophils,as well as the activation of the PI3K-AKT signaling pathway,better reflect the favorable conditions and clinical significance of effective recanalization.(3)The simulation of transient occlusion of the middle cerebral artery in rats with cerebral ischemia and intravascular mechanical thrombectomy suggests that early intravascular mechanical thrombectomy improves neurological function and infarct size after cerebral ischemia.Further detection results suggest that natural killer cells,neutrophils,important target DEm Rs,and PI3K-AKT signaling pathways are involved in the treatment process of mechanical thrombectomy in AIS patients.Targeted immunoinflammatory therapy may be an effective strategy for assisting intravascular mechanical thrombectomy in the treatment of cerebral ischemia.