Tourniquet-induced Skeletal Muscle Research Articles

Inflammatory Responses of the Remote Organs After Tourniquet-Induced Ischemia-Reperfusion in Mouse Hindlimb

Tourniquet use is the most effective tool for controlling life-threatening extremity hemorrhage before other treatments and for creating bloodless operating fields in surgical procedures. However, tourniquet-induced ischemia-reperfusion (IR) also causes skeletal muscle injury and associated secondary end-organ damage. Our previous studies have demonstrated that there are involvements of inflammatory cytokines in tourniquet-induced skeletal muscle IR injuries. The objective of this study is to investigate the inflammatory responses of end-organs after tourniquet-induced mouse hindlimb IR. We hypothesized that tourniquet use would result in an inflammatory response in the remote organs. The unilateral hindlimbs of mice were subjected to 3 hours of tourniquet application by placing a rubber band at the hip joint. Then the rubber bands were released for reperfusion over different periods of time (1, 3, 7, and 14 days). The mRNA levels of IL-1β and TNFα (two inflammatory cytokines) in the lungs, liver, kidneys, and heart were measured using real-time RT-PCR analysis. In the lungs, the levels of IL-1β and TNFα mRNA were significantly elevated at day 1 after tourniquet-induced IR, then decreased at days 3 and 7, and returned to a high level at day 14. In the liver, the levels of IL-1β and TNFα mRNA were increased at day 1 and remained at a high level until day 14. However, tourniquet-induced hindlimb IR did not change the levels of IL-1β or TNFα mRNA in the kidneys or heart during the whole period of tourniquet-induced IR. In conclusion, these results suggest that the inflammatory response to tourniquet-induced hindlimb IR is different among distant organs, which will help to develop therapeutic strategies for different end-organs after tourniquet-induced hindlimb IR. NIH/NIGMS 1R01GM145736 to Y-LL. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Effect of dexmedetomidine on tourniquet-induced skeletal muscle injury.

The aim of this study was to investigate whether dexmedetomidine could reduce tourniquet-induced skeletal muscle injury. C57BL6 male mice were randomly assigned to sham, ischemia/reperfusion, and dexmedetomidine groups. Mice in the ischemia/reperfusion and dexmedetomidine groups received normal saline solution and dexmedetomidine intraperitoneally, respectively. The sham group underwent the same procedure as the ischemia/reperfusion group, with the exception of tourniquet application. Subsequently, the ultrastructure of the gastrocnemius muscle was observed, and its contractile force was examined. In addition, Toll-like receptor 4 and nuclear factor-κB expression within muscles was detected by Western blot. Dexmedetomidine alleviated myocyte damage and increased the contractility of skeletal muscles. Moreover, dexmedetomidine significantly inhibited the expression of Toll-like receptor 4/nuclear factor-κB in the gastrocnemius muscle. Taken together, these results demonstrate that dexmedetomidine administration attenuated tourniquet-induced structural and functional impairment of the skeletal muscle, partly through inactivation of the Toll-like receptor 4/nuclear factor-κB pathway.

The Effect of Air Tourniquet on Interleukin-6 Levels in Total Knee Arthroplasty.

Background:Air tourniquet-induced skeletal muscle injury increases the concentrations of some cytokines such as interleukin-6 (IL-6) in plasma. However, the effect of an air tourniquet on the IL-6 concentrations after total knee arthroplasty (TKA) is unclear. We therefore investigated the impact of tourniquet-induced ischemia and reperfusion injury in TKA using the IL-6 level as an index.Methods:Ten patients with primary knee osteoarthrosis who underwent unilateral TKA without an air tourniquet were recruited (Non-tourniquet group). We also selected 10 age- and sex-matched control patients who underwent unilateral TKA with an air tourniquet (Tourniquet group). Venous blood samples were obtained at 3 points; before surgery, 24 h after surgery, and 7 days after surgery.The following factors were compared between the two groups; IL-6, C-reactive protein (CRP), creatine phosphokinase (CPK), the mean white blood cell (WBC) counts, and the maximum daily body temperatures.Results:The IL-6 level at 24 h after surgery was significantly higher than that at any other point (p<0.01). No significant differences were observed in the WBC count, the body temperature, or the CRP, CPK, or IL-6 levels of the two groups at any of the time points.Conclusion:The effect of ischemia and reperfusion due to the use of an air tourniquet on increasing the IL-6 level was much smaller than that induced by surgical stress in TKA.

Protective Effects of Sonic Hedgehog Against Ischemia/Reperfusion Injury in Mouse Skeletal Muscle via AKT/mTOR/p70S6K Signaling

Background/Aims: Skeletal muscle ischemia/reperfusion (I/R) injury is a common and severe disease. Sonic hedgehog (Shh) plays a critical role in post-natal skeletal muscle regeneration. In the present study, the role of Shh in skeletal muscle I/R injury and the mechanisms involved were investigated. Methods: The expression of Shh, AKT/mTOR/p70S6K and apoptosis pathway components were evaluated following tourniquet-induced skeletal muscle I/R injury. Then, mice were subjected to systemic administration of cyclopamine or one-shot treatment of a plasmid encoding the human Shh gene (phShh) to examine the effects of Shh on I/R injury. Moreover, mice were subjected to systemic administration of NVP-BEZ235 to investigate the role of the AKT/mTOR/p70S6K pathway in Shh-triggered skeletal muscle protection. Results: We found that the levels of Shh, AKT/mTOR/p70S6K pathway components and Cleaved Caspase 3 and the Bax/Bcl2 ratio initially increased and then decreased at different time points post-I/R injury. Moreover, Shh protected skeletal muscle against I/R injury by alleviating muscle destruction, reducing interstitial fibrosis and inhibiting apoptosis, and these protective effects were abrogated when the AKT/mTOR/p70S6K pathway was inhibited. Conclusion: Collectively, these data suggest that Shh signaling exerts a protective role through the AKT/mTOR/p70S6K signaling pathway during skeletal muscle I/R injury. Thus, Shh signaling may be a therapeutic target for protecting skeletal muscle from I/R injury.

Administration of FTY720 during Tourniquet-Induced Limb Ischemia Reperfusion Injury Attenuates Systemic Inflammation.

Acute ischemia-reperfusion injury (IRI) of the extremities leads to local and systemic inflammatory changes which can hinder limb function and can be life threatening. This study examined whether the administration of the T-cell sequestration agent, FTY720, following hind limb tourniquet-induced skeletal muscle IRI in a rat model would attenuate systemic inflammation and multiple end organ injury. Sprague-Dawley rats were subjected to 1 hr of ischemia via application of a rubber band tourniquet. Animals were randomized to receive an intravenous bolus of either vehicle control or FTY720 15 min after band placement. Rats (n = 10/time point) were euthanized at 6, 24, and 72 hr post-IRI. Peripheral blood as well as lung, liver, kidney, and ischemic muscle tissue was analyzed and compared between groups. FTY720 treatment markedly decreased the number of peripheral blood T cells (p < 0.05) resulting in a decreased systemic inflammatory response and lower serum creatinine levels and had a modest but significant effect in decreasing the transcription of injury-associated target genes in multiple end organs. These findings suggest that early intervention with FTY720 may benefit the treatment of IRI of the limb. Further preclinical studies are necessary to characterize the short-term and long-term beneficial effects of FTY720 following tourniquet-induced IRI.

Use of dextran sulfate in tourniquet-induced skeletal muscle reperfusion injury

BackgroundLower extremity ischemia–reperfusion injury (IRI)—prolonged ischemia and the subsequent restoration of circulation—may result from thrombotic occlusion, embolism, trauma, or tourniquet application in surgery. The aim of this study was to assess the effect of low-molecular-weight dextran sulfate (DXS) on skeletal muscle IRI. MethodsRats were subjected to 3 h of ischemia and 2 or 24 h of reperfusion. To induce ischemia the femoral artery was clamped and a tourniquet placed under the maintenance of the venous return. DXS was injected systemically 10 min before reperfusion. Muscle and lung tissue samples were analyzed for deposition of immunoglobulin M (IgM), IgG, C1q, C3b/c, fibrin, and expression of vascular endothelial-cadherin and bradykinin receptors b1 and b2. ResultsAntibody deposition in reperfused legs was reduced by DXS after 2 h (P < 0.001, IgM and IgG) and 24 h (P < 0.001, IgM), C3b/c deposition was reduced in muscle and lung tissue (P < 0.001), whereas C1q deposition was reduced only in muscle (P < 0.05). DXS reduced fibrin deposits in contralateral legs after 24 h of reperfusion but did not reduce edema in muscle and lung tissue or improve muscle viability. Bradykinin receptor b1 and vascular endothelial-cadherin expression were increased in lung tissue after 24 h of reperfusion in DXS-treated and non-treated rats but bradykinin receptor b2 was not affected by IRI. ConclusionsIn contrast to studies in myocardial infarction, DXS did not reduce IRI in this model. Neither edema formation nor viability was improved, whereas deposition of complement and coagulation components was significantly reduced. Our data suggest that skeletal muscle IRI may not be caused by the complement or coagulation alone, but the kinin system may play an important role.

Tourniquet-induced acute ischemia–reperfusion injury in mouse skeletal muscles: Involvement of superoxide

Although arterial limb tourniquet is one of the first-line treatments to prevent exsanguinating hemorrhage in both civilian pre-hospital and battlefield casualty care, prolonged application of a limb tourniquet can lead to serious ischemia–reperfusion injury. However, the underlying pathomechanisms of tourniquet-induced ischemia–reperfusion injury are still poorly understood. Using a murine model of acute limb ischemia–reperfusion, we investigated if acute limb ischemia–reperfusion injury is mediated by superoxide overproduction and mitochondrial dysfunction. Hind limbs of C57/BL6 mice were subjected to 3 h ischemia and 4 h reperfusion via placement and release of a rubber tourniquet at the greater trochanter. Approximately 40% of the gastrocnemius muscle suffered infarction in this model. Activities of mitochondrial electron transport chain complexes including complex I, II, III, and IV in the gastrocnemius muscle were decreased in the ischemia–reperfusion group compared to sham. Superoxide production was increased while activity of manganese superoxide dismutase (MnSOD, the mitochondria-targeted SOD isoform) was decreased in the ischemia–reperfusion group compared to the sham group. Pretreatment with tempol (a SOD mimetic, 50 mg/kg) or co-enzyme Q 10 (50 mg/kg) not only decreased the superoxide production, but also reduced the infarct size and normalized mitochondrial dysfunction in the gastrocnemius muscle. Our results suggest that tourniquet-induced skeletal muscle ischemia–reperfusion injuries including infarct size and mitochondrial dysfunction may be mediated via superoxide overproduction and reduced antioxidant activity. In the future, this murine ischemia–reperfusion model can be adapted to mechanistically evaluate anti-ischemic molecules in tourniquet-induced skeletal muscle injury.

Influence of fiber-type composition on recovery from tourniquet-induced skeletal muscle ischemia–reperfusion injury

This study was designed to determine if previously reported differences in the functional impairment of muscles composed of predominantly different fiber types occurs following extended periods of ischemia. We hypothesized that the soleus (Sol) muscle, a predominantly slow-twitch muscle, would be less vulnerable to tourniquet-induced ischemia-reperfusion than the plantaris (Plant), a predominantly fast-twitch muscle, as determined by the assessment of isometric contractile function. Male Sprague-Dawley rats were assigned to one of the following groups to undergo tourniquet application (TKA) (n = 6/group): 2 h TKA, 2 d recovery; 4 h TKA, 2 d recovery; 2 h TKA, 14 d recovery; or 4 h TKA, 14 d recovery. In situ isometric contractile properties were assessed in the predominantly slow-twitch Sol and the predominantly fast-twitch Plant; the contralateral muscle served as the internal control. At 2 d, muscle contraction could not be elicited via neural stimulation, but muscles did contract with direct stimulation, which indicates neural injury. This condition was resolved by day 14. At this time point, tetanic tension (Po) in the Plant was reduced by 45% and 69% in the 2 and 4 h groups, respectively. Po for the Sol was unaffected in the 2 h group, but was reduced by 30% in the 4 h group. The fatigue resistance of the Plant was increased 2 fold in the 4 h group and was unchanged in all other groups. These results demonstrate that vulnerability to tourniquet-induced ischemia-reperfusion injury is dramatically different with respect to muscle fiber-type composition.

Sevoflurane but not propofol increases interstitial glycolysis metabolites availability during tourniquet-induced ischaemia–reperfusion

Ischaemia/reperfusion (I/R) is one of the main pathophysiological phenomena involved in the anaesthetic practice. The authors hypothesized that anaesthetic regimens can influence skeletal muscle tolerance to tourniquet-induced I/R that should be reflected by the interstitial metabolite levels of anaerobic glycolysis. Microdialysis probes were implanted in three groups of 10 patients each receiving either sevoflurane (SEVO), propofol (PRO), or spinal (SA) anaesthesia (for induction and maintenance). SA group was considered as a control group. Interstitial fluid was obtained during tourniquet-induced I/R and was analysed for interstitial glucose, lactate, pyruvate, and glycerol. The microdialysis flow rate was 0.5 microl min(-1). Compared with the control group, the SEVO group had a higher level of both lactate and pyruvate and an increase in glucose during ischaemia. In contrast, the PRO group had a lower level of pyruvate, resulting in a significant higher increase (eight times from baseline) of the lactate pyruvate ratio. Glucose level remained low in this group. During reperfusion, lactate, pyruvate, and glucose remained at a significantly higher level in the SEVO group. In the PRO group, there was no difference in lactate, pyruvate, and glucose levels compared with the control group. The interstitial level of glycerol exhibits only few and comparable changes during I/R between the groups. Our results indicate that there is a better availability of interstitial glycolysis metabolites (glucose, lactate, and pyruvate) in the skeletal muscle during ischaemia and reperfusion after sevoflurane exposure than after propofol, suggesting a potential preconditioning effect of sevoflurane on tourniquet-induced skeletal muscle I/R.

Influence of Systemic Hypotension on Skeletal Muscle Ischemia-Reperfusion Injury After 4-Hour Tourniquet Application

Tourniquet use for extremity hemorrhage control is common in military trauma. Tourniquet use may be accompanied by systemic hypotension, but this phenomenon has not been studied. We aimed to define the muscle effects of the combined insult of tourniquet-induced skeletal muscle ischemia-reperfusion injury (I-R) and hemorrhagic hypotension. After a 33% carotid arterial hemorrhage, Sprague-Dawley rats underwent 240-min hindlimb ischemia induced by pneumatic tourniquet. Control animals were not hemorrhaged. No resuscitation was given. After tourniquet release, muscles were reperfused for 120 min and then dissected. Weights and mitochondrial viability assays (nitroblue tetrazolium method) were performed on the plantaris (PL), and soleus (SO). Histologic analysis was performed on the PL and SO. Muscle edema is expressed as the ratio of tourniquet limb to contralateral limb muscle weight. Animal laboratories of the United States Army Institute of Surgical Research. STUDY ANIMALS: Twelve Sprague-Dawley rats. The mean arterial pressure of hemorrhaged animals was 38 +/- 3 mm Hg before tourniquet placement and 86 +/- 4 mm Hg before release, both significantly (p < 0.05) lower than controls at the same time points. Pre- tourniquet mortality was 38% with hemorrhage and 0% without. All muscles experienced edema, with weight ratios greater than 1. The PL experienced significantly (p < 0.05) less edema with hemorrhage. Viability was unaffected by hemorrhage in all muscles, as was tissue inflammation. No differences in inflammation were observed with hemorrhage. Systemic hypotension modulates the impact of 4 hours of tourniquet ischemia by decreasing muscle edema but minimally altering measures of muscle viability. Compartment anatomy and muscle fiber type both influence muscle response to the combined insult of hypotension and I-R. In this model, hypotension did not worsen the skeletal muscle I-R observed after the use of a tourniquet for 4 hours.

Fluid resuscitation following hemorrhage influences acute tourniquet-induced skeletal muscle ischemia-reperfusion injury

Journal of the American College of Surgeons 201(3):p S51, September 2005. | DOI: 10.1016/j.jamcollsurg.2005.06.111

Effects of an Antioxidant in a Rabbit Model of Tourniquet-Induced Skeletal Muscle Ischemia–Reperfusion Injury

The purpose of this study was to evaluate the effects of U74006F, a new antioxidant, on isometric contractile function after tourniquet-induced ischemia–reperfusion injury. Twenty-six NZW rabbits underwent either 2 or 4 hr of thigh tourniquet compression. Animals were randomized to receive U74006F or an equal volume of its citrate vehicle. When tibialis anterior function was tested 2 days later, there was no difference between treatment groups in peak tension, rate of force production, contraction time, half relaxation time, or resistance to fatigue. The use of U74006F failed to improve functional outcome in this model of ischemia–reperfusion injury.