Ischemic preconditioning upregulates Mitofusin2 and preserves muscle strength in tourniquet-induced ischemia/reperfusion

Prangmalee Leurcharusmee; Passakorn Sawaddiruk; Yodying Punjasawadwong; Nantawit Sugandhavesa; Kasisin Klunklin; Siam Tongprasert; Patraporn Sitilertpisan; Nattayaporn Apaijai; Nipon Chattipakorn; Siriporn C Chattipakorn

doi:10.1016/j.jot.2022.09.012

Ischemic preconditioning upregulates Mitofusin2 and preserves muscle strength in tourniquet-induced ischemia/reperfusion

J Orthop Translat. 2022 Oct 14:35:113-121. doi: 10.1016/j.jot.2022.09.012. eCollection 2022 Jul.

Authors

Prangmalee Leurcharusmee^{1

2

3}, Passakorn Sawaddiruk^{1

2

3}, Yodying Punjasawadwong³, Nantawit Sugandhavesa⁴, Kasisin Klunklin⁴, Siam Tongprasert⁵, Patraporn Sitilertpisan⁶, Nattayaporn Apaijai^{1

2

7}, Nipon Chattipakorn^{1

2

7}, Siriporn C Chattipakorn^{1

2

8}

Affiliations

¹ Neurophysiology Unit, Cardiac Electrophysiology Research and Training Center, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
² Center of Excellence in Cardiac Electrophysiology Research, Chiang Mai University, Chiang Mai, Thailand.
³ Department of Anesthesiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
⁴ Department of Orthopedics, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
⁵ Department of Rehabilitation, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
⁶ Faculty of Associated Medical Science, Chiang Mai University, Chiang Mai, Thailand.
⁷ Cardiac Electrophysiology Unit, Department of Physiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand.
⁸ Department of Oral Biology and Diagnostic Sciences, Faculty of Dentistry, Chiang Mai University, Chiang Mai, Thailand.

Abstract

Background: Tourniquet-induced ischemia and reperfusion (I/R) has been related to postoperative muscle atrophy through mechanisms involving protein synthesis/breakdown, cellular metabolism, mitochondrial dysfunction, and apoptosis. Ischemic preconditioning (IPC) could protect skeletal muscle against I/R injury. This study aims to determine the underlying mechanisms of IPC and its effect on muscle strength after total knee arthroplasty (TKA).

Methods: Twenty-four TKA patients were randomized to receive either sham IPC or IPC (3 cycles of 5-min ischemia followed by 5-min reperfusion). Vastus medialis muscle biopsies were collected at 30 min after tourniquet (TQ) inflation and the onset of reperfusion. Western blot analysis was performed in muscle protein for 4-HNE, SOD2, TNF-ɑ, IL-6, p-Drp1^ser616, Drp1, Mfn1, Mfn2, Opa1, PGC-1ɑ, ETC complex I-V, cytochrome c, cleaved caspase-3, and caspase-3. Clinical outcomes including isokinetic muscle strength and quality of life were evaluated pre- and postoperatively.

Results: IPC significantly increased Mfn2 (2.0 ± 0.2 vs 1.2 ± 0.1, p = 0.001) and Opa1 (2.9 ± 0.3 vs 1.9 ± 0.2, p = 0.005) proteins expression at the onset of reperfusion, compared to the ischemic phase. There were no differences in 4-HNE, SOD2, TNF-ɑ, IL-6, p-Drp1^ser616/Drp1, Mfn1, PGC-1ɑ, ETC complex I-V, cytochrome c, and cleaved caspase-3/caspase-3 expression between the ischemic and reperfusion periods, or between the groups. Clinically, postoperative peak torque for knee extension significantly reduced in the sham IPC group (-16.6 [-29.5, -3.6] N.m, p = 0.020), while that in the IPC group was preserved (-4.7 [-25.3, 16.0] N.m, p = 0.617).

Conclusion: In TKA with TQ application, IPC preserved postoperative quadriceps strength and prevented TQ-induced I/R injury partly by enhancing mitochondrial fusion proteins in the skeletal muscle.

The translational potential of this article: Mitochondrial fusion is a potential underlying mechanism of IPC in preventing skeletal muscle I/R injury. IPC applied before TQ-induced I/R preserved postoperative quadriceps muscle strength after TKA.

Keywords: 4-HNE, 4-hydroxy-2-nonenal; ADP, Adenosine diphosphate; ASA, American Society of Anesthesiologists; ATP, Adenosine triphosphate; BSA, Bovine serum albumin; CAT, Catalase; CHOP, C/EBP homologous protein; Drp1, Dynamin-related protein-1; ER, Endoplasmic reticulum; ETC, Electron transport chain; FGF21, Fibroblast growth factor 21; Fis1, Fission protein-1; GPx, Glutathione peroxidase; I/R, Ischemia and reperfusion; IL-6, Interleukin-6; IPACK, Interspace between the popliteal artery and capsule of the posterior knee; IPC, Ischemic preconditioning; Ischemia reperfusion injury; Ischemic preconditioning; Knee arthroplasty; MDA, Malondialdehyde; Mfn, Mitofusin; Mitochondrial dynamics; MnSOD, Manganese superoxide dismutase; NF-κB, Nuclear factor kappa B; OXPHOS, Oxidative phosphorylation; PGC-1ɑ, Peroxisome proliferator-activated receptor-gamma coactivator-1ɑ; RIPC, Remote ischemic preconditioning; ROS, Reactive oxygen species; SBP, Systolic blood pressure; SOD, Superoxide dismutase; TKA, Total knee arthroplasty; TNF, Tumor necrosis factor; TQ, Tourniquet; Tourniquet; UPR, Unfolded protein response; mPTP, Mitochondrial permeability transition pore.