Effect of different anaesthetic techniques on gene expression profiles in patients who underwent hip arthroplasty

Renata Alleva, Andrea Tognù, Marco Tomasetti, Maria Serena Benassi, Laura Pazzaglia, Hanna van Oven, Ettore Viganò, Nicola De Simone, Ilaria Pacini, Sandra Giannone, Sanjin Gagic, Raffaele Borghi, Sara Picone, Battista Borghi, Renata Alleva, Andrea Tognù, Marco Tomasetti, Maria Serena Benassi, Laura Pazzaglia, Hanna van Oven, Ettore Viganò, Nicola De Simone, Ilaria Pacini, Sandra Giannone, Sanjin Gagic, Raffaele Borghi, Sara Picone, Battista Borghi

Abstract

Objectives: To investigate the modulation of genes whose expression level is indicative of stress and toxicity following exposure to three anaesthesia techniques, general anaesthesia (GA), regional anaesthesia (RA), or integrated anaesthesia (IA).

Methods: Patients scheduled for hip arthroplasty receiving GA, RA and IA were enrolled at Rizzoli Orthopaedic Institute of Bologna, Italy and the expression of genes involved in toxicology were evaluated in peripheral blood mononuclear cells (PBMCs) collected before (T0), immediately after surgery (T1), and on the third day (T2) after surgery in association with biochemical parameters.

Results: All three anaesthesia methods proved safe and reliable in terms of pain relief and patient recovery. Gene ontology analysis revealed that GA and mainly IA were associated with deregulation of DNA repair system and stress-responsive genes, which was observed even after 3-days from anaesthesia. Conversely, RA was not associated with substantial changes in gene expression.

Conclusions: Based on the gene expression analysis, RA technique showed the smallest toxicological effect in hip arthroplasty.

Trial registration: ClinicalTrials.gov number NCT03585647.

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1. Overview of study design according…
Fig 1. Overview of study design according to CONSORT format.
Accordantly to eligibility, 130 patients were enrolled. Of them, 31 patients were excluded from the study, then 99 patients were randomised in the three anaesthesia groups.
Fig 2. Gene expression analysis in patients…
Fig 2. Gene expression analysis in patients receiving general anaesthesia (GA).
(A) Heatmap of significant gene expression in PBMCs of patients (n = 3) undergoing elective hip arthroplasty receiving GA, immediately after operation (T1) and on the third postoperative day (T2), adjusted at p-value less than 0.05. Genes with greater and lower abundance after anaesthesia (FC, fold change) are shown in red and green, respectively, with significance highlighted in yellow. Normalization and microarray analysis were performed by RT2 Profiler PCR Array Analysis software version 3.5 (SABiosciences). (B) Gene-gene interaction analysis of the significantly differentially expressed genes showing the networks of deregulated pathways at T2 time point.
Fig 3. Gene expression analysis in patients…
Fig 3. Gene expression analysis in patients receiving regional anaesthesia (RA).
(A) Heatmap of significant gene expression in PBMCs of patients (n = 3) undergoing elective hip arthroplasty receiving RA, immediately after operation (T1) and on the third postoperative day (T2), adjusted at p-value less than 0.05. Genes with greater and lower abundance after anaesthesia (FC, fold change) are shown in red and green, respectively, with significance highlighted in yellow. Normalization and microarray analysis were performed by RT2 Profiler PCR Array Analysis software version 3.5 (SABiosciences). (B) Gene-gene interaction analysis of the significantly differentially expressed genes showing the networks of deregulated pathways at T2 time point.
Fig 4. Gene expression analysis in patients…
Fig 4. Gene expression analysis in patients receiving integrated anaesthesia (IA).
(A) Heatmap of significant gene expression in PBMCs of patients (n = 3) undergoing elective hip arthroplasty receiving IA, immediately after operation (T1) and on the third postoperative day (T2), adjusted at p-value less than 0.05. Genes with greater and lower abundance after anaesthesia (FC, fold change) are shown in red and green, respectively, with significance highlighted in yellow. Normalization and microarray analysis were performed by RT2 Profiler PCR Array Analysis software version 3.5 (SABiosciences). (B) Gene-gene interaction analysis of the significantly differentially expressed genes showing the networks of deregulated pathways at T2 time point.
Fig 5. Venn diagram and gene clustering.
Fig 5. Venn diagram and gene clustering.
(A) Venn diagram that shows differentially expressed genes that were shared among three anaesthesia methods. Cluster analysis of differentially expressed genes in general anaesthesia (Group-1), regional anaesthesia (Group-2), and integrated anaesthesia (Group-3) at third postoperative day (T2), respect to control Group (before anaesthesia, T0). (B) Gene clustering by their expression levels.
Fig 6. Expression of significantly deregulated stress-responsive…
Fig 6. Expression of significantly deregulated stress-responsive genes.
Expression of DNA repair proteins (ERCC1, ERCC3, RAD50, XRCC1, XRCC2), early growth response protein 1 (EGR1), heat shock factor 1 (HSF1), glutathione peroxidase (GPX), and superoxide dismutase (SOD2) in patients undergoing arthroplasty before anaesthesia (T0), immediately after operation (T1), and third postoperative day (T2). General anaesthesia (GA, n = 25), regional anaesthesia (RA, n = 25), integrated anaesthesia (IA, n = 25). The data shown are expressed as fold change at T1 and T2 with respect to T0 time points. The symbol ‘*’ denotes significant differences in data at T1 and T2 versus T0 time points, with symbol ‘°’ significance between data at T1 and T2 time points with p<0.05.

References

    1. Barroso Rosa S, James D, Matthews BD. Is knee arthroscopy under local anaesthetic a patient-friendly technique? A prospective controlled trial. Eur J Orthop Surg Traumatol. 2016; 26: 633–638. 10.1007/s00590-016-1799-2
    1. Kratz T, Dette F, Schmitt J, Wiesmann T, Wulf H, Zoremba M. Impact of regional femoral nerve block during general anaesthesia for hip arthroplasty on blood pressure, heart rate and pain control: A randomized controlled study. Technol Health Care. 2015; 23: 313–22. 10.3233/THC-150898
    1. Borghi B, Casati A, Iuorio S, Celleno D, Michael M, Serafini PL, et al. Effect of different anaesthesia techniques on red blood cell endogenous recovery in hip arthroplasty. J Clin Anesth. 2005; 17: 96–101. 10.1016/j.jclinane.2004.05.005
    1. Liszka H, Gądek A. Pre-emptive Local Anaesthesia in Ankle Arthroscopy. Foot Ankle Int. 2016; 37: 1326–1332. 10.1177/1071100716665354
    1. Kumara AB, Gogia AR, Bajaj JK, Agarwal N. Clinical evaluation of post-operative analgesia comparing suprascapular nerve block and interscalene brachial plexus block in patients undergoing shoulder arthroscopic surgery. J Clin Orthop Trauma. 2016;7: 34–39. 10.1016/j.jcot.2015.09.003
    1. Lowes DA, Galley HF, Moura AP, Webster NR. Brief isoflurane anaesthesia affects differential gene expression, gene ontology and gene networks in rat brain. Behav Brain Res. 2017; 317: 453–460. 10.1016/j.bbr.2016.09.045
    1. Pan JZ, Wei H, Hecker JG, Tobias JW, Eckenhoff RG, Eckenhoff MF. Rat brain DNA transcript profile of halothane and isoflurane exposure. Pharmacogenet Genomics. 2006; 16: 171–182. 10.1097/01.fpc.0000189795.21770.08
    1. Sakamoto A, Imai J, Nishikawa A, Honma R, Ito E, Yanagisawa Y, et al. Influence of inhalation anaesthesia assessed by comprehensive gene expression profiling. Gene. 2005;356: 39–48. 10.1016/j.gene.2005.03.022
    1. Lee ML, Whitmore GA. Power and sample size for DNA microarray studies. Stat Med. 2002;21: 3543–3570. 10.1002/sim.1335
    1. Chen LH, Liang J, Chen MC, Wu CC, Cheng HS, Wang HH, et al. The relationship between preoperative American Society of Anesthesiologists Physical Status Classification scores and functional recovery following hip-fracture surgery. BMC Musculoskelet Disord. 2017;18: 410 10.1186/s12891-017-1768-x
    1. Tomasetti M, Alleva R, Borghi B, Collins AR. In vivo supplementation with coenzyme Q10 enhances the recovery of human lymphocytes from oxidative DNA damage. FASEB J. 2001;15: 1425–1427. 10.1096/fj.00-0694fje
    1. Yu D, Huang LJ, Chen NM. Anesthetic Propofol-Induced Gene Expression Changes in Patients Undergoing Coronary Artery Bypass Graft Surgery Based on Dynamical Differential Coexpression Network Analysis. Comput Math Methods Med. 2016;2016: 7097612.
    1. Yamashita K, Matsumoto H, Saito F, Takeyoshi M. Differences in gene expression profiles in liver caused by different types of anaesthesia: cases of CO2-O2 and isoflurane. J Toxicol Sci. 2015;40: 829–836. 10.2131/jts.40.829
    1. Edmands SD, Ladow E, Hall AC. Microarray Analyses of Genes Regulated by Isoflurane Anaesthesia In Vivo: A Novel Approach to Identifying Potential Preconditioning Mechanisms. Anesth Analg. 2013;116: 589–595. 10.1213/ANE.0b013e31827b27b0
    1. Culley DJ, Yukhananov RY, Xie Z, Gali RR, Tanzi RE, Crosby G. Altered hippocampal gene expression 2 days after general anaesthesia in rats. Eur J Pharmacol. 2006;549: 71–78. 10.1016/j.ejphar.2006.08.028
    1. Lowes DA, Galley HF, Moura AP, Webster NR. Brief isoflurane anaesthesia affects differential gene expression, gene ontology and gene networks in rat brain. Behav Brain Res. 2017;317: 453–460. 10.1016/j.bbr.2016.09.045
    1. Xu Z, Lu Y, Wang J, Ding X, Chen J, Miao C. The protective effect of propofol against TNF-α-induced apoptosis was mediated via inhibiting iNOS/NO production and maintaining intracellular Ca2+ homeostasis in mouse hippocampal HT22 cells. Biomed Pharmacother. 2017;91: 664–672. 10.1016/j.biopha.2017.04.110
    1. Yu X, Sun X, Zhao M, Hou Y, Hou Y, Li J, et al. Propofol attenuates myocardial ischemia reperfusion injury partly through inhibition of resident cardiac mast cell activation. Int Immunopharmacol. 2018;54: 267–274. 10.1016/j.intimp.2017.11.015
    1. Yoon JY, Jeon HO, Kim EJ, Kim CH, Yoon JU, Park BS et al. Propofol protects human keratinocytes from oxidative stress via autophagy expression. J Dent Anesth Pain Med. 2017;17: 21–28. 10.17245/jdapm.2017.17.1.21
    1. Szyfter K, Stachecki I, Kostrzewska-Poczekaj M, Szaumkessel M, Szyfter-Harris J, Sobczyński P. Exposure to volatile anaesthetics is not followed by a massive induction of single-strand DNA breaks in operation theatre personnel. J Appl Genet. 2016;57: 343–348. 10.1007/s13353-015-0329-y
    1. Xu Z, Yu J, Wu J, Qi F, Wang H, Wang Z, et al. The Effects of Two Anesthetics, Propofol and Sevoflurane, on Liver Ischemia/Reperfusion Injury. Cell Physiol Biochem. 2016;38: 1631–1642. 10.1159/000443103
    1. Zhang Y, Chen Z, Feng N, Tang J, Zhao X, Liu C et al. Protective effect of propofol preconditioning on ischemia-reperfusion injury in human hepatocyte. Thorac Dis. 2017;9: 702–710.
    1. Alleva R, Tomasetti M, Solenghi MD, Stagni F, Gamberini F, Bassi A, et al. Lymphocyte DNA damage precedes DNA repair or cell death after orthopaedic surgery under general anaesthesia. Mutagenesis. 2003;18: 423–428. 10.1093/mutage/geg013
    1. Souza KM, Braz LG, Nogueira FR, Souza MB, Bincoleto LF, Aun AG, et al. Occupational exposure to anesthetics leads to genomic instability, cytotoxicity and proliferative changes. Mutat Res. 2016;791–792: 42–48. 10.1016/j.mrfmmm.2016.09.002
    1. Ni C, Li C, Dong Y, Guo X, Zhang Y, Xie Z. Anesthetic Isoflurane Induces DNA Damage Through Oxidative Stress and p53 Pathway. Mol Neurobiol. 2017;54: 3591–3605. 10.1007/s12035-016-9937-8
    1. Nogueira FR, Braz LG, de Andrade LR, de Carvalho AL, Vane LA, Módolo NS, et al. Evaluation of genotoxicity of general anaesthesia maintained with desflurane in patients under minor surgery. Environ Mol Mutagen. 2016;57: 312–316. 10.1002/em.22012
    1. Grishko V, Xu M, Wilson G, Pearsall AW 4th. Apoptosis and mitochondrial dysfunction in human chondrocytes following exposure to lidocaine, bupivacaine, and ropivacaine. J Bone Joint Surg Am. 2010;92: 609–618. 10.2106/JBJS.H.01847
    1. Dragoo JL, Braun HJ, Kim HJ, Phan HD, Golish SR. The in vitro chondrotoxicity of single-dose local anesthetics. Am J Sports Med. 2012;40: 794–799. 10.1177/0363546511434571
    1. Pines A, Bivi N, Romanello M, Damante G, Kelley MR, Adamson ED, et al. Cross-regulation between Egr-1 and ape/ref-1 during early response to oxidative stress in the human osteoblastic hobit cell line: Evidence for an autoregulatory loop. Free Radic Res. 2005;39: 269–281. 10.1080/10715760400028423
    1. Pagel JI, Deindl E. Disease Progression Mediated by Egr-1 Associated Signaling in Response to Oxidative Stress. Int J Mol Sci. 2012;13: 13104–13117. 10.3390/ijms131013104
    1. Hasan RN, Phukan S, Harada S. Differential regulation of early growth response gene-1 expression by insulin and glucose in vascular endothelial cells. Arterioscler Thromb Vasc Biol. 2003;23: 988–993. 10.1161/01.ATV.0000071351.07784.19
    1. Dayalan Naidu S, Kostov RV, Dinkova-Kostova AT. Transcription factors Hsf1 and Nrf2 engage in crosstalk for cytoprotection. Trends in Pharmacol Sci. 2015; 36: 6–14.
    1. Rosenfeldt F, Wilson M, Lee G, Kure C, Ou R, Braun L, et al. Oxidative stress in surgery in an ageing population: pathophysiology and therapy. Exp Gerontol. 2013;48: 45–54. 10.1016/j.exger.2012.03.010
    1. Bech NH, Hulst AH, Spuijbroek JA, van Leuken LL, Haverkamp D. Perioperative pain management in hip arthroscopy; what options are there? J Hip Preserv Surg. 2016;3: 181–189. 10.1093/jhps/hnw015

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