Role And Mechanism Of Propofol On The Regulation Of Postoperative Immune Function And Inflammatory Response In Patients With Lung Cancer

Chapter 1 Study of Propofol-regulating Macrophages Function in Lung squamous cell carcinomaBackgrounds: Lung cancer is one of the most common cause of cancer-related death worldwide.Previous studies showed that propofol shows immunological characteristics related to tumor immune microenvironment in the occurrence and development of non-small cell lung cancer.However,the effect of propofol on immune function of patients with Lung squamous cell carcinoma undergoing surgical resection has not been reported.Objective: To verify the potential regulation mechanism of propofol on regulating macrophages,this study would explore the impact of propofol on immune function of patients with Lung squamous cell carcinoma undergoing surgical resection.Methods: The Gene Expression Omnibus(GEO)database of the National Center for Biotechnology Information was used to search the mRNA sequencing data of Lung squamous cell carcinoma and adjacent tissues based on single cell sequencing,and use R(version: x6.4.3)software to analyze whole data.Then,we combined with the previously published prior knowledge related to the immune system to explore all genes related to various immune response processes.The deferentially expressed genes in macrophages of tumor tissues and paracancerous tissues,and different cell types were identified via t-SNE,UMAP and Principal Component Analysis.The gene function analysis annotation tools DAVID was used to analyze gene ontology and KEGG signaling pathway of the differentially expressed genes,for the identification of downstream signaling pathways and possible biological processes.Forty lung cancer patients were randomly divided into propofol total intravenous anesthesia group(P group)and sevoflurane inhalation anesthesia group(S group)using random number expression.Finally,RT-qPCR was used to detect the node genes of related signaling pathways in the exosomes of patients with lung squamous cell carcinoma before and after resection.Results:1.The first 30 principal components were included in the follow-up analysis,and 854 cells were divided into 12 subgroups.The 12 subgroups were annotated according to the expression of marker gene in different peripheral blood cells in Cellmarker database,and the subgroups under the same category were combined.Finally,seven kinds of cell annotation were obtained,and the number of epithelial cells(4 v.s 228),B cells(3 v.s 58),macrophages(25 v.s 60)in cancer tissue was significantly reduced than that in the adjacent tissues.2.To analyze the gene differential expression of macrophages between cancer tissue and non-cancer tissue,a total of 739 deferentially expressed genes were obtained.Go analysis showed that the deferentially expressed genes in macrophages were mainly enriched in protein translation,antigen processing and presentation and other immune functions.KEGG analysis showed that the most abundant signal pathways were autophagosome and lysosome,which may be involved in the regulation of macrophage autophagy.3.Such autophagy related genes as Atg9 a,Atg3,Atg7,Atg13 and Atg14 play a key node role.The expression levels of Atg9 a and Atg3 were significantly decreased after operation under propofol anesthesia.Conclusion: Macrophage autophagy plays a key role in the development of Lung squamous cell carcinoma.Propofol anesthesia would improve postoperative immune function through the activation of macrophage autophagy.Chapter 2 Role of propofol on the immune function regulation in lung adenocarcinoma based on miRNAObjective: To investigate the effect of propofol and sevoflurane on the expression of miRNA and mRNA in lung adenocarcinoma,and to explore the mechanism of propofol regulating immune function based on miRNA expression.Methods: The TCGA database was used to search miRNA and mRNA expression profile data in lung adenocarcinoma tissues and non-cancer tissues,and use Affy and Limma packages of R(version: x6.4.3)software to to explore the differentially expressed miRNAs that were involved in the regulation of immune function and signal pathway.We evaluated the miRNAs and the nodal molecules regulating the signaling pathway in the exosomes of patients undergoing lung adenocarcinoma resection with propofol(P group)or sevoflurane(S group)anesthesia via RT-qPCR.Using the online miRNA target gene prediction website Target Scan to identify the target genes of DE-miRNAs,and the gene function analysis annotation tools DAVID to analyze gene ontology and KEGG signaling pathway of the target genes.The association between miRNA involved in the regulation of immune function and prognosis were analyzed.Results:1.A total of 265 differential expressed miRNAs including 155 upregulated miRNAs and 110 downregulated miRNAs were identified.Simultaneously,a total of 4261 differential expressed mRNAs including 2362 upregulated mRNAs and 1899 downregulated mRNAs were identified.2.According to the unsupervised hierarchical clustering analysis,the differentially expressed mRNA was divided into 10 different modules.The GO analysis of the genes set of module 6 showed that the genes of this module were involved in the immune processes such as neutrophil activation,T cell activation and lymphocyte proliferation.The expression of miRNA sets in modules 1,15 and 33 were correlated with that in module 6.3.The expression levels of miR-30d-5p,miR-30b-3p,miR-200a-5p,miR-3934-3p and miR-101-3p were correlated with the overall survival time of patients,but no miRNA was correlated with tumor progression.4.There were no significant differences noted in the relative expression of miR-200b-3p,miR-30b-5p,miR-30d-5p,miR-519c-5p and miR-455-5p between the P and S groups before and after operation.However,the expressions of miR-200a-5p and miR-516a-5p in exosomes of group P were higher than those of group S(both P < 0.05).5.There were 154 target genes predicted by miR-200a-5p.These target genes of miR-200a-5p are mainly involved in regulation of transcription,DNAtemplated,mRNA splicing via spliceosome,protein modification by small protein conjugation,positive regulation of endothelial cell proliferation,GPI anchor biosynthetic process and other biological processes;such cellular component as intracellular,U4/U6 x U5 tri-sn RNP complex and nuclear speck;molecular function as nucleic acid binding,dolichyl-phosphate-glucoseglycolipid alpha-glucosyltransferase activity and DNA binding.KEGG analysis showed these target genes of miR-200a-5p were enriched in spliceosome,endocytosis and herpes simplex infection.6.There were 154 target genes predicted by miR-516a-5p.The target genes of miR-516a-5p are mainly involved in negative regulation of transcription fromRNA polymerase II promoter,embryonic foregut morphogenesis,regulation of vesicle fusion,stimulatory C-type lectin receptor signaling pathway and other biological processes;such cellular component as nucleolus,extrinsic component of Golgi membrane,endoplasmic reticulum and Golgi apparatus;molecular function as GTPase activator activity,transcription factor activity,sequence-specific DNA binding,acetylcholine receptor inhibitor activity and sequence-specific DNA binding.KEGG analysis showed these target genes of miR-200a-5p were enriched in p53 signaling pathway,calcium signaling pathway,Chagas disease(American trypanoso-miasis),thyroid hormone signaling pathway and long-term potentiation.Conclusion:Compared with sevoflurane,propofol would be more preferable to improve cellular immune function through targeting on miRNA-200a-5p and miRNA-516a-5p.Chapter 3 Effects of Propofol versus Sevoflurane on Postoperative Inflammation and Prognosis in Patients with Non-Small Cell Lung CancerObjective: To investigate the effects and mechanisms of propofol versus sevoflurane anesthesia on inflammatory response and prognosis after thoracoscopic radical resection of non-small cell lung cancer(NSCLC).Methods: A totoal of 61 patients with NSCLC were randomly divided into propofol based total intravenous anesthesia group(P group)and sevoflurane based inhalation anesthesia group(S group).The serum samples were collected from patients in both groups at each time points during perioperative period.The levels of such immune cells as neutrophils,T lymphocytes,B lymphocytes and NK cells determined by flow cytometry(FCM).The levels of cytokines such as IL-1β,IL-2,IL-6 and IL-121 in serum were determined by enzymy-linked immunosorbent assay(ELISA)to evaluate the inflammation and immune status during perioperative period.Cardiopulmonary function indexes(such as heart rate,mean arterial pressure),liver and kidney functions(such as AST,ALT,serum creatinine)were measured in each group to understand the perioperative physiological conditions of patients.Such survival data as recurrence and survival time were collected.To understand the effects of propofol and sevoflurane on the survival of NSCLC patients,Cox proportional risk model was constructed with SPSS version 23.0 to analyze patients’ survival after surgery.Results:1.Compared with the P group,the recovery time and extubation time were both prolonged(both P < 0.05).The mean recovery time of the P and S groups were both less than 90 min,but the mean recovery time in the S group was significantly prolonged by 20 min than that in the P group.There were 7(23%)patients in the P group and 9(29%)patients in the S group undergoing delayed recovery.The mean extubation time in the S group was significantly prolonged by 30 min than that in the P group.However,the mean hospitalization stay in the S group was lower by 1.2 d than that in the P group.Such perioperative data as anesthesia time,operation time,transfusion volume,urine volume and use of vasoactive drugs were similar between these two groups(both P > 0.05).2.The MAP at O-L 1h and T-L 30 min in the P group were both increased in comparison with the S group(both P < 0.05),but the MAP at each time points between these two group were both less than 105 mm Hg.The HR at O-L 1h and T-L 30 min in the P group were both decreased in comparison with the S group(both P < 0.05),but the MAP at each time points between these two group were both less than 80 beats/min.3.NEU counts in the S group at postoperative 7 days was higher with1.33×109/L than that in the P group,but these variation is within the normal range(P = 0.008).Compared with the P group,the LYM counts in the S group at postoperative 1 days was decreased while that at postoperative 3 and 7 days were increased(both P < 0.05).There were no statistical differences noted on the CD3~+T lymphocytes,CD4~+T lymphocytes and CD19+B lymphocytes at each time points between these two group(both P > 0.05).Compared with the P group,CD8~+T lymphocytes and NK cells counts in the S group at postoperative7 days were both decreased(both P < 0.05),but the Th/Ts ratio at postoperative3 and 7 days in the S group were both increased(both P < 0.05).4.There were no statistical differences noted on the IL-β and IL-2 level at each time points between these two group(both P > 0.05).The serum IL-6 level at 1 and 7 days after operation in the P group were both higher than that in the S group(both P < 0.05),while the serum IL-12 level at postoperative 3 days in the P group were increased(P = 0.043).5.There were no statistical differences noted on the ALT,AST,urea and Ccr levels at each time points between these two group(both P > 0.05).There were no statistical differences noted on the creatinine levels at baseline and the time after operation between these two group(both P > 0.05),but the creatinine levels at 1 days after operation in the P group were decreased in comparison with the S group(P = 0.012).Among the included patients,there were 8 patients respectively in the P and S group whose AST levels at at 1 days after operation were more than 40 U/L.6.The chest radiographs of these two groups showed local lesions without obvious abnormality in the remaining lung lobe and tracheobronchial.The incidence of pleural effusion in the P group was significantly higher than that of the S Group at 7 days after operation.Conclusion:Clinical study showed that sevoflurane anesthesia is more valuable to affect the function of CD8~+ T cells and NK cells.