Quantitative Proteomic Analysis of Plasma after Remote Ischemic Conditioning in a Rhesus Monkey Ischemic Stroke Model

Abstract

Background: Animal and clinical studies have shown that remote ischemic conditioning (RIC) has protective effects for cerebral vascular diseases, with induced humoral factor changes in the peripheral blood. However, many findings are heterogeneous, perhaps due to differences in the RIC intervention schemes, enrolled populations, and sample times. This study aimed to examine the RIC-induced changes in the plasma proteome using rhesus monkey models of strokes. Methods: Two adult rhesus monkeys with autologous blood clot-induced middle cerebral artery (MCA) occlusion underwent RIC interventions twice a week for five consecutive weeks. Each RIC treatment included five cycles of five minutes of ischemia alternating with five minutes of reperfusion of the forearm. The blood samples were taken from the median cubital vein of the monkeys at baseline and immediately after each week’s RIC stimulus. The plasma samples were isolated for a proteomic analysis using mass spectrometry (MS). Results: Several proteins related to lipid metabolism (Apolipoprotein A-II and Apolipoprotein C-II), coagulation (Fibrinogen alpha chain and serpin), immunoinflammatory responses (complement C3 and C1), and endovascular hemostasis (basement membrane-specific heparan sulfate proteoglycan) were significantly modulated after the RIC intervention. Many of these induced changes, such as in the lipid metabolism regulation and anticoagulation responses, starting as early as two weeks following the RIC intervention. The complementary activation and protection of the endovascular cells occurred more than three weeks postintervention. Conclusions: Multiple protective effects were induced by RIC and involved lipid metabolism regulation (anti-atherogenesis), anticoagulation (antithrombosis), complement activation, and endovascular homeostasis (anti-inflammation). In conclusion, this study indicates that RIC results in significant modulations of the plasma proteome. It also provides ideas for future research and screening targets.

Remote ischemic conditioning-induced hyperacute and acute responses of plasma proteome in healthy young male adults: a quantitative proteomic analysis

“…Hepponstall et al [19] have demonstrated that ApoA-I, A-II, and D levels are upregulated during the early response (15 min) and late response (24 h) phases after one-time RIC stimulation in healthy young adults. Previously, we showed that long-term use of RIC might elevate the ApoA-II level in the primate ischemic stroke model [18] . Thus, we speculated that the induced elevation of multiple apolipoproteins post-RIC might be consistent with the effects of lipid metabolism regulation by repeated RIC stimulations.…”

“…Samples were separated through a gradient of 5–31% ACN in 0.1% FA (buffer B) on the ReproSil-Pur Basic C18 column (120 mm × 150 μm, 1.9 μm particle size; Dr. Maisch HPLC GmbH, Ammerbuch, Germany) at 250 nL/min flow rate. The MS parameter setup was performed as described in our previous studies [18] …”

“…Long-term intervention (5 weeks, twice a week) of RIC in the experimental AIS rhesus model has shown activation of complement, endovascular homeostasis, anti-coagulation mechanism, and regulation of lipid metabolism (anti-atherogenesis). [ 18 ] Only one study focused on the post-RIC effect in healthy male adults, with the exclusion of probable proteomic changes under a certain diseased condition. [ 19 ] Hepponstall et al [ 19 ] have demonstrated that a single RIC could upregulate fibrinogen beta chain (FGB), fibrinogen gamma (FGG), and apolipoprotein A (ApoA)-mediating signaling as early as 15 min after RIC, and sustain up to 24 h. However, the inflation/deflation pressures of RIC were not clearly mentioned in the study.…”

Potential Anti-Inflammatory and Anti-Coagulation Effects of One-Time Application of Remote Ischemic Conditioning in Patients With Subacute/Chronic Cerebral Arteriostenosis and Venostenosis

“…Besides, we further illustrated the underlying mechanisms that RIC could decrease stroke recurrence and white matter lesions by improving cerebral perfusion, reducing inflammation via decreasing C-reactive protein (CRP), interleukin-6 (IL-6), leukocyte count, and inhibiting the coagulation process through decreasing plasminogen activator inhibitor-1 (PAI-1) and platelet aggregation rate in patients with intracranial atherosclerotic stenosis 11. Moreover, we conducted a proteomic analysis to examine the long-term effects of RIC (5-wk consistent stimulus) in our rhesus monkey ischemic stroke model 14. The coagulation process was modulated by down-regulating levels of fibrinogen alpha chain and serpin.…”

“…11 Moreover, we conducted a proteomic analysis to examine the long-term effects of RIC (5-wk consistent stimulus) in our rhesus monkey ischemic stroke model. 14 The coagulation process was modulated by down-regulating levels of fibrinogen alpha chain and serpin.…”