The application of angiogenesis in the lower limb ischemia test is an important mechanism for improving blood perfusion after ischemic cardiovascular disease blood flow obstruction. However, there is currently no clinically simple, non-invasive and specific method for direct evaluation of angiogenesis. The lower limb ischemia test is an important test for the evaluation of angiogenesis. The method is to prepare a lower limb ischemia model by using a rat or a mouse to ligature the lower extremity femoral artery, and the other lower limb is used as a control group. The posterior limb blood flow was analyzed by laser Doppler flow imaging. The hind limbs were then scanned and imaged weekly to analyze the recovery of blood flow in the hind limbs.

The lower limb ischemia test is an important method for the study of ischemic cardiovascular disease and therapeutic angiogenesis. The angiogenic factor can be injected into the body to promote ischemic local angiogenesis and collateral circulation, and improve the blood supply and function of the tissue. Further, the effect of promoting angiogenic factors was evaluated. Proangiogenic factors can be pro-angiogenic drugs, proteins, genes, and stem cell therapies.

In addition to cardiovascular disease research, angiogenesis studies in diabetes, peripheral vascular disease, wound healing, etc. can also be evaluated using the lower limb ischemia test.

Required equipment

MoorLDI2-HIR point scanning laser Doppler flow imaging (Gene&I, Beijing) AMG small animal anesthesia machine (Gene&I, Beijing)

Shaving knife and hair removal cream

Experimental procedure

The rats or mice were anesthetized using an anesthesia machine.

The hair removal cream is applied to the hind limbs.

Cut one side of the hind limb skin and separate the muscle to separate the femoral artery.

The femoral artery is ligated in close proximity to the groin using a suture.

The wound was sutured and a blood flow scan image of both lower extremities was performed using a laser Doppler scanner.

The blood flow of the lower limbs and the normal side of the ischemic side was analyzed.

test results

On the day after surgery, the ischemic rate of the ischemic limb relative to the normal side can reach 80-90%.

After the use of pro-angiogenic factors, the blood flow recovery process of the ischemic limb can be observed.

Reference

Aicher A., ​​Heeschen C., Sasaki K., Urbich C., Zeiher AM, Dimmeler S. 2006

Low-energy shock wave for enhancing recruitment of endothelial progenitor cells: a new modality to increase efficacy of cell therapy in chronic hind limb ischemia.

Circulation, 114 (25), pp 2823-30

Chalothorn D, Zhang H, Clayton J. A, Thomas S. A, Faber J. E, 2005.

Catecholamines augment collateral vessel growth and angiogenesis in hindlimb ischemia.

American Journal of Physiology - Heart and Circulatory Physiology Published 1 August 2005 Vol. 289 no. 2, H947-H959.

Crawford RS, Albadawi H., Atkins MD, Jones JE, Yoo HJ, Conrad MF, Austen Jr WG, Watkins MT 2010

Postischemic poly (ADP-ribose) polymerase (PARP) inhibition reduces ischemia reperfusion injury in a hind-limb ischemia model.

Surgery, 148 (1), pp 110 - 118

Tritsaris K., Myren M., Ditlev SB, Hübschmann MV, van der Blom I., Hansen AJ, Olsen UB, Cao R., Zhang J., Jia T., Wahlberg E., Dissing S., Cao Y. 2007

L-20 is an arteriogenic cytokine that remodels collateral networks and improves functions of ischemic hind limbs.

Proc Natl AcadSci US A.104 (39), pp 15364-9

Turgeon J., Dussault S., Haddad P., Groleau J., Ménard C, Michaud SE, Maingrette F., Rivard A. 2010

Probucol and antioxidant Vitamins rescue ischemia-induced neovascularization .......... exposed to cigarette smoke: Potential role of endothelial progenitor cells.

Atherosclerosis 208, spp 342–349

Ji'an Deer Lower Limb Ischemia Domestic Cooperation Unit

Six Academy of Military Medical Sciences

Fudan University School of Basic Medicine

East China Normal University

Peking University School of Medicine

Institute of Biophysics, Chinese Academy of Sciences

West China Institute of Regenerative Medicine, Sichuan University

Sichuan Medical University