MicroRNA-126 mimic administration accelerates vascular perfusion recovery and angiogenesis in a hind limb ischemia model

Abstract Background Peripheral arterial disease caused mainly by atherosclerosis portent significant morbidity, adverse prognosis and mortality, with localized treatment approaches aiming at symptom alleviation and improvement of circulation. Recently, scientific interest has been shifted towards epigenomics, with microRNAs appearing as a future therapeutic target in ischemic cardiovascular diseases due to their potential in regulating angiogenesis. Purpose We investigated the pro-angiogenic effect of miRNA-126 mimic in an in vivo model of hind limb ischemia. Methods Ten-week-old male C57Bl/6 mice (n=20) were subjected to left femoral artery ligation and were treated with microRNA-126 mimic at a dose of 5mg/kg (Group A, n=10) or 0.2ml normal saline (Group B, n=10) on days 1, 3 and 7. Laser Doppler imaging was performed to verify successful ligation on day 0 and to evaluate differences in the ischemic-to-normal (I/N) hind limb perfusion ratio on day 7 and 28. Muscle tissue expression of microRNA-126 and vascular endothelial growth factor (VEGF) was determined via PCR. Results Following microRNA-126 mimic administration in Group A subjects, we noted a qualitative and quantitative stepwise increase in I/N hind limb perfusion ratio [Day 0: 0.354 (0.276, 0.455) vs. Day 8: 0.775 (0.700, 0.844) vs. Day 28: 0.681 (0.660, 0.896), p=0.001] (Figure 1, Panels A and B). In Group B a stepwise increase of lesser magnitude was observed in I/N hind limb perfusion ratio [Day 0: 0.267 (0.164, 0.383) vs. Day 8: 0.400 (0.338, 0.418) vs. Day 28: 0.539 (0.483, 0.603), p=0.074]. Importantly, over time changes of I/N hind limb perfusion ratio were significantly higher in group A compared to group B (p for interaction=0.005) (Figure 1, Panel B). Muscle tissue expression of microRNA-126 in the ischemic hind limb of Group A was 350-fold lower compared to the ischemic hind limb of Group B (p<0.001) (Figure 1, Panel C). A higher expression (14.2-fold) of VEGF in the ischemic hind limb of microRNA-126-treated mice compared to that of control group was detected (p<0.001) (Figure 1, Panel C). A statistically significant negative correlation was noted between microRNA-126 and VEGF tissue expression levels in the ischemic limbs of both Group A and B subjects whereas no correlation between microRNA-126 and VEGF was observed in the non-ischemic hind limbs of the entire study population (Figure 1, Panel D). Conclusion MicroRNA-126 mimic delivery in the ischemic hind limb of mice can accelerate vascular perfusion recovery via angiogenesis, which is mediated by VEGF expression. FUNDunding Acknowledgement Type of funding sources: None. Figure 1

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The Effect of MicroRNA-126 Mimic Administration on Vascular Perfusion Recovery in an Animal Model of Hind Limb Ischemia

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.724465 ◽

2021 ◽

Vol 8 ◽

Author(s):

Panagiotis Theofilis ◽

Georgia Vogiatzi ◽

Evangelos Oikonomou ◽

Maria Gazouli ◽

Gerasimos Siasos ◽

...

Keyword(s):

Muscle Tissue ◽

Hind Limb ◽

Tissue Expression ◽

Control Group ◽

Vascular Perfusion ◽

Entire Study ◽

Stepwise Increase ◽

Limb Perfusion ◽

Group A ◽

Group B

Background: MicroRNAs have been linked to angiogenesis and could prove to be valuable future therapeutic targets in ischemic cardiovascular diseases.Methods: Ten-week-old male C57Bl/6 mice were subjected to left femoral artery ligation and were treated with microRNA-126 mimic at a dose of 5 mg/kg (Group A, n = 10) or 5 mg/kg microRNA mimic negative control (Group B, n = 10) on days 1, 3, and 7. Laser Doppler imaging was performed to verify successful ligation on day 0 and to evaluate differences in the ischemic-to-normal (I/N) hind limb perfusion ratio on day 28. Muscle tissue expression of microRNA-126 and vascular endothelial growth factor (VEGF) was determined via PCR.Results: Following microRNA-126 mimic administration in Group A subjects, we noted a stepwise increase in I/N hind limb perfusion ratio (Day 0: 0.364 ± 0.032 vs. Day 8: 0.788 ± 0.049 vs. Day 28: 0.750 ± 0.039, p = 0.001). In Group B a stepwise increase in I/N hind limb perfusion ratio was observed (Day 0: 0.272 ± 0.057 vs. Day 8: 0.382 ± 0.020 vs. Day 28: 0.542 ± 0.028, p = 0.074). Muscle tissue expression of microRNA-126 in the ischemic hind limb of Group A was 350-fold lower compared to the ischemic hind limb of Group B (p < 0.001). A higher expression (14.2-fold) of VEGF in the ischemic hind limb of microRNA-126-treated mice compared to that of control group was detected (p < 0.001). A statistically significant negative correlation was noted between microRNA-126 and VEGF tissue expression levels in the ischemic limbs of the entire study population.Conclusion: MicroRNA-126 delivery in the ischemic hind limb of mice improved vascular perfusion with VEGF upregulation.

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Risk factors for critical limb ischemia in patients undergoing femoral cannulation for venoarterial extracorporeal membrane oxygenation: Is distal limb perfusion a mandatory approach?

Perfusion ◽

10.1177/0267659119827231 ◽

2019 ◽

Vol 34 (6) ◽

pp. 453-459 ◽

Cited By ~ 6

Author(s):

Tim Kaufeld ◽

Eric Beckmann ◽

Fabio Ius ◽

Nurbol Koigeldiev ◽

Wiebke Sommer ◽

...

Keyword(s):

Risk Factors ◽

Risk Factor ◽

Extracorporeal Membrane Oxygenation ◽

Limb Ischemia ◽

Distal Limb ◽

Membrane Oxygenation ◽

Venoarterial Extracorporeal Membrane Oxygenation ◽

Limb Perfusion ◽

Group A ◽

Group B

Background: Venoarterial extracorporeal membrane oxygenation support is a well-established tool in the care of severe refractory cardiac and respiratory failure. The application of this support may serve as a bridge to transplant, recovery or to implantation of a ventricular assist device. Venoarterial extracorporeal membrane oxygenation support can be administered through an open surgical access via the common femoral or axillary artery or a percutaneous approach using Seldinger technique. Both techniques may obstruct the blood flow to the lower limb and may cause a significant ischemia with possible limb loss. Malperfusion of the distal limb can be avoided using an ipsilateral distal limb perfusion, which may be established by adding a single-lumen catheter during venoarterial extracorporeal membrane oxygenation treatment to overcome the obstruction. The aim of this study is to distinguish the presence or absence of a distal limb perfusion regarding the incidence of distal limb ischemia. Furthermore, expected risk factors of open and percutaneous femoral venoarterial extracorporeal membrane oxygenation installation were evaluated for the development of distal limb ischemia. Methods: Between January 2012 and September 2015, 489 patients received venoarterial extracorporeal membrane oxygenation support at our institution. In total, 307 patients (204 male, 103 female) with femoral cannulation were included in the analysis. The cohort was distinguished by the presence (group A; n = 237) or absence (group B; n = 70) of a distal limb perfusion during peripheral venoarterial extracorporeal membrane oxygenation treatment. Furthermore, a risk factor analysis for the development of distal limb ischemia was performed. Results: The main indications for venoarterial extracorporeal membrane oxygenation therapy were a low cardiac output syndrome (LCOS) (53%) and failed weaning of extracorporeal circulation (23%). A total of 23 patients (7.49%) under venoarterial extracorporeal membrane oxygenation support developed severe distal limb malperfusion (3.38% in group A vs 21.42% in group B). Preemptive installation of distal limb perfusion extended the intervention-free intervals to 7.8 ± 19.3 days in group A and 6.3 ± 12.5 in group B. A missing distal limb perfusion (p = 0.001) was identified as a main risk factor for critical limb ischemia. Other comorbidities such as arterial occlusion disease (p = 0.738) were not statistically significantly associated. Surgical intervention due to vascular complications after extracorporeal membrane oxygenation explantation was needed in 14 cases (4.22% in group A and 5.71% in group B). Conclusion: We were able to identify the absence of distal limb perfusion as an independent risk factor for the development of critical distal limb ischemia during femoral venoarterial extracorporeal membrane oxygenation treatment. The application of a distal limb perfusion should be considered as a mandatory approach in the context of femoral venoarterial extracorporeal membrane oxygenation treatment regardless of the implantation technique.

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Danhong Promotes Angiogenesis in Diabetic Mice after Critical Limb Ischemia by Activation of CSE-H2S-VEGF Axis

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2015/276263 ◽

2015 ◽

Vol 2015 ◽

pp. 1-8 ◽

Cited By ~ 3

Author(s):

Feng Wu ◽

Zhiqing He ◽

Ru Ding ◽

Zhigang Huang ◽

Qixia Jiang ◽

...

Keyword(s):

Critical Limb Ischemia ◽

Hind Limb ◽

Limb Ischemia ◽

Saline Control ◽

Diabetic Patients ◽

Hind Limb Ischemia ◽

Beneficial Effects ◽

Danhong Injection ◽

Limb Perfusion ◽

Endothelial Growth Factor

The aim of this paper is to investigate effect and mechanism of Danhong injection (DH) on angiogenesis in the diabetic hind limb ischemia mouse model. Thirty diabetic hind limb ischemic model mice and ten normal mice, established by intraperitoneal (i.p.) injection of streptozotocin (STZ) or PBS and ligation/excision of femoral artery, and then twenty diabetic hind limb ischemic model mice of all were evenly randomized to saline (control,n=10) and DH i.p. injection (2 mL/kg weight for 7 days,n=10) groups. Limb perfusion recovery and femoral blood hydrogen sulfide (H2S) and vessel regeneration and lower limb vascular endothelial growth factor (VEGF)/cystathionineγ-lyase (CSE) expression were evaluated during intervention and after euthanasia, respectively. DH i.p. increased ischemic limb perfusion and promoted collateral circulation generation without decreasing blood glucose level. Increased local CSE-H2S-VEGF expression contributed to beneficial effects of DH injection. In conclusion, activation of local CSE-H2S-VEGF axis might participate in proangiogenesis effects of DH injection in diabetic hind limb ischemia model mice, suggesting a potential therapy for diabetic patients with critical limb ischemia.

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PC232. Vitamin D 3 -Induced Arterial Calcification Decreases Functional Recovery and Limb Perfusion in a Murine Model of Hind Limb Ischemia

Journal of Vascular Surgery ◽

10.1016/j.jvs.2016.03.374 ◽

2016 ◽

Vol 63 (6) ◽

pp. 223S-224S

Author(s):

Sara L. Zettervall ◽

Stephanie Monk ◽

Xue-Lin Wang ◽

Tonghui Lin ◽

Yujun Cai ◽

...

Keyword(s):

Vitamin D ◽

Functional Recovery ◽

Murine Model ◽

Hind Limb ◽

Limb Ischemia ◽

Arterial Calcification ◽

Hind Limb Ischemia ◽

Limb Perfusion ◽

Vitamin D 3

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Faculty Opinions recommendation of Soluble CD146 displays angiogenic properties and promotes neovascularization in experimental hind-limb ischemia.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.3963958.3722058 ◽

2010 ◽

Author(s):

Lindsey A Miles ◽

Shahrzad Lighvani

Keyword(s):

Hind Limb ◽

Limb Ischemia ◽

Hind Limb Ischemia ◽

Soluble Cd146

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FACS-purified myoblasts producing controlled VEGF levels induce safe and stable angiogenesis in chronic hind limb ischemia

Journal of Cellular and Molecular Medicine ◽

10.1111/j.1582-4934.2011.01308.x ◽

2011 ◽

Vol 16 (1) ◽

pp. 107-117 ◽

Cited By ~ 16

Author(s):

Thomas Wolff ◽

Edin Mujagic ◽

Roberto Gianni-Barrera ◽

Philipp Fueglistaler ◽

Uta Helmrich ◽

...

Keyword(s):

Hind Limb ◽

Limb Ischemia ◽

Hind Limb Ischemia

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WITHDRAWN: Melatonin attenuates lung injury in a hind limb ischemia–reperfusion rat model

Revista Portuguesa de Pneumologia ◽

10.1016/j.rppneu.2014.01.007 ◽

2014 ◽

Author(s):

Hamed Takhtfooladi ◽

Mohammad Takhtfooladi ◽

Fariborz Moayer ◽

Sayed Mobarakeh

Keyword(s):

Lung Injury ◽

Rat Model ◽

Hind Limb ◽

Ischemia Reperfusion ◽

Limb Ischemia ◽

Hind Limb Ischemia

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A Modified Surgical Model of Hind Limb Ischemia in ApoE-/- Mice using a Miniature Incision

Journal of Visualized Experiments ◽

10.3791/62402 ◽

2021 ◽

Author(s):

Kaixuan Yan ◽

Jiaxing Zheng ◽

Frank G. Zöllner ◽

Kay Schwenke ◽

Prama Pallavi ◽

...

Keyword(s):

Hind Limb ◽

Limb Ischemia ◽

Hind Limb Ischemia ◽

Surgical Model

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Abstract 17597: Endogenous Del-1 is a Negative Regulator of Ischemia-induced Angiogenesis by Interacting With the Leukocytic LFA-1 Integrin

Circulation ◽

10.1161/circ.130.suppl_2.17597 ◽

2014 ◽

Vol 130 (suppl_2) ◽

Author(s):

Sebastian Cremer ◽

Anne Klotzsche-von Ameln ◽

Alessia Orlandi ◽

Irina Korovina ◽

Bettina Gercken ◽

...

Keyword(s):

Endothelial Cell ◽

Hind Limb ◽

Hematopoietic Cells ◽

Limb Ischemia ◽

Inflammatory Cells ◽

Capillary Density ◽

Negative Regulator ◽

Hind Limb Ischemia ◽

Autonomous Pathway

Developmental endothelial locus-1 (Del-1) is an endothelial cell-derived secreted protein circulating in blood and associated with the cell surface and the extracellular matrix. As we previously demonstrated, Del-1 restricts leukocyte recruitment by inhibiting the β2-integrin, LFA-1. Leukocytes and progenitor cells (PC) may contribute to angiogenesis. The role of endogenous Del-1 in angiogenesis is elusive. We found, that physiological angiogenesis of the developing retina was not affected in the Del-1-/- mice compared to the wildtype (WT) mice. Surprisingly, Del-1-/- mice displayed a significantly increased angiogenic response compared to WT mice after induction of hind limb ischemia (144 ± 6 % increase of capillary density) and retinal ischemia (retinopathy of prematurity model) suggesting that endogenous Del-1 is an inhibitor of ischemia-induced neovascularization. Silencing of Del-1 with siRNA did not affect the angiogenic sprouting of endothelial cell (EC) spheroids, indicating that Del-1 blocks angiogenesis in a non-endothelial cell autonomous pathway. Soluble Del-1 blocked the adhesion of inflammatory cells on EC monolayers. In line with these results, ischemic muscles and ischemic retinae from Del-1-/- mice displayed an enhanced infiltration with inflammatory cells compared to the WT mice. Since Del-1 blocks inflammatory cell homing by inhibiting the leukocytic LFA-1-integrin, we addressed the role of the Del-1/LFA-1-integrin interaction on the inhibitory function of endogenous Del-1 on angiogenesis. Indeed, Del-1/LFA-1-double deficiency reversed the pro-angiogenic phenotype of the Del-1-/- mice to the level of WT mice in the model of hind limb ischemia. Thus, the inhibitory role of Del-1 on neovascularization is mediated by the interaction of Del-1 with the LFA-1-integrin. Moreover, Del-1-deficiency led to an increased homing of intravenously injected murine fluorescence-labeled WT Lin- BM PC in ischemic muscles in comparison to WT mice after the induction of hind limb ischemia. Taken together, Del-1 acts as a negative regulator of ischemia-induced angiogenesis by interacting with the LFA-1-integrin expressed in hematopoietic cells, thereby inhibiting the homing of hematopoietic cells to ischemic tissues.

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