OBJECTIVETo investigate the best injection method of the bone marrow mesenchymal stem cells (BM-MSCs) transplantation for the treatment of a rat model with hind limb ischemia.

METHODSTwenty four SD rats with hind limb ischemia were randomly divided into four groups: control group, model group, acupoint BM-MSCs injection group (API group) and thigh muscle BM-MSCs injection group (TMI group). The acupoints of "Sanyinjiao" (SP 6), "Housanli" (ST 36), "Zhaohai" (KI 6), "Huantiao" (GB 30) and "Yanglingquan" (GB 34) were selected for API group, and five non-acupoints were selected on gastrocnemius and adductor of ischemic hind limb for TMI group. Both groups were accepted BM-MSCs transplantion. Model rat with hind limb ischemia was established with the method of blocking the femoral artery and its branches. The changes of blood flow (perfuse unit, PU) was monitored with laser Doppler flowmetry (LDF). In order to describe the visual changes in blood flow, the PU index (PUI) was determined as the ratio of ischemic to non-ischemic hind limb blood perfusion. And also, the levels of VEGF,bFGF in serum were tested to analyze the immunohistochemical expression quantity of VEGF and bFGF.

RESULTSComparing with the model and the TMI groups, the PUI value on 3rd, 14th and 21th days after BM-MSCs transplantation were significantly increased in the API group (P < 0.05, P < 0.01). In contrast to the model group, the VEGF,bFGF levels in serum and the immunohistochemical expression quantity of VEGF and bFGF in the API and TMI groups were significantly increased (all P < 0.01).

CONCLUSIONTransplantation of BM-MSCs through the acupoint can more significantly and quickly increase the blood flow and cause the greater improvement on hind limb ischemia than that of through the way of muscle injection.