粒细胞集落刺激因子在自体外周血干细胞移植治疗缺血性下肢血管病中的作用

Role of granulocyte colony stimulating factor in the transplantation of autologus peripheral blood stem cells for treatment of ischemic lower-limb angionosis

目的:观察缺血性下肢血管病患者进行自体外周血干细胞移植时,应用粒细胞集落刺激因子后细胞成分的变化以及对自身身体状况的近期影响。方法:选取2004-11/2005-04解放军第四六三医院内分泌科收治的126例接受粒细胞集落刺激因子动员的缺血性下肢血管病患者,全部接受皮下注射粒细胞集落刺激因子5～12μg/(kg·d),连续4～5d。为防止血黏度增加引起心脑血管意外,在干细胞动员的同时应用低分子肝素钙5000u,皮下注射1次/d,连续4～5d。每天监测血细胞计数和凝血3项,同时采用流式细胞仪监测外周血中CD34+细胞数。观察并记录动员后及采集过程中、后出现的毒副反应。结果:按意向处理分析,实验纳入126例缺血性下肢血管病患者,全部进入结果分析。①全部患者动员过程中外周血象的变化:粒细胞集落刺激因子动员前白细胞数量为(5.35±1.64)×109L-1,动员第5天为(42.17±18.56)×109L-1,第6天为(44.23±17.47)×109L-1,动员后比动员前提高5～13倍(P<0.01);血红蛋白和血小板动员前后无明显变化。②全部患者动员后采集细胞悬液的情况:37例患者于动员第5天进行采集,单个核细胞数值和CD34+百分数分别为(432.68±89.36)×109L-1和(0.87±0.38)%;其余89例均于第6天进行采集,单个核细胞数值和CD34+百分数分别为(463.71±58.33)×109L-1和(0.90±0.35)%,两者基本相近(P>0.05)。③性别、年龄和体质量对单个核细胞数值和CD34+细胞百分数的影响:男性患者采集的单个核细胞数值高于女性患者(P<0.05),而单位体质量的CD34+细胞数值男女基本相近;以年龄55岁为界,大于55岁和小于55岁的患者差异显著(P<0.05);高体质量患者采集的单个核细胞数值高于低体质量患者(P<0.05),而单位体质量的CD34+细胞数值基本相似。④不良事件和副反应:主要的不良反应有骨痛、周身肌肉酸痛、乏力、头痛、失眠、食欲下降、恶心呕吐、低热,采集过程中可能出现口周、面部或四肢麻木,一般停药2～4d症状即可消失。结论:缺血性下肢血管病患者进行自体外周血干细胞移植时,粒细胞集落刺激因子作为有效动员剂,可有效动员单个核细胞和CD34+细胞,绝大部分患者能够耐受,但应使用一定剂量的抗凝剂预防不良反应的发生。

AIM： To observe the changes of cellular composition in lower-limb isehemic angionosis patients who got autologns peripheral blood stem cell transplantation with granulocyte colony stimulating factor （G-CSF） and the short-term influence of G-CSF on the health of the patients. METHODS： We chosen 126 patients who suffered lower limb ischemic angionosis and received the treatment with G-CSF in the Department of Endocrinology, the 463 Hospital of Chinese PLA from November 2004 to April 2005. All the patients received the subcutaneous administration of G-CSF of 5-12 μg/（kg·d）, for 4 to 5 successive days . In order to avoid the cardiovascular disease caused by the increase of blood viscosity, 5 000 u lower molecular heparin calcium was subcutaneouslly injected, once a day, for 4 to 5 successive days. Blood cell count and blood coagulation were detected every day, meanwhile , the number of CD34^＋ in the peripheral blood was detected with flow cytometer. Side effect of G-CSF was observed during and after sample collection. RESULTS： According to intention-to-treat analysis, totally 126 patients with lower limb ischemic angionosis were enrolled in the experiment and all of them entered the stage of result analysis. ① Change of peripheral blood of the whole process of all the patients： the number of white blood cell （WBC） was （5,35±1,64）×10^9 L^-1 before G-CSF was applied and it was （42.17±18.56）×10^9 L^-1 on the 5^th day and （44.23±17.47）×10^9 L^-1 on the 6^th day. The number of WBC was enhanced 5 to 13 times that before G-CSF was used （P〈 0.01）; There was no obvious change of the peripheral blood before and after hemoglobin and platelet were used. ② Cell suspension of all the patients： on the 5^th day after G-CSF was used in 37 patients, peripheral blood mononuclear cells and CD34^＋ percent was （432.68±89.36）×10^9 L^-1 and （0.87±0.38）%, respectively ; the other 89 cases were detected on the 6^th day , peripheral blood mononuclear cells and CD34^＋ percent was （463.71 ±58.33）×10^9 L^-1 and （0.90±0.35）% , respectively, both of them were similar （P 〉 0.05）. ③Effect of gender, age and body mass on mononuclear cells and CD34^＋ percent： the number of peripheral blood mononuclear cells from male patients was higher than that from female patients （P 〈 0.05） ,there was no significant difference of CD34^＋ number in unit body mass ; 55 years as the borderline , there was significant difference in those aged over 55 years and less than 55 years （P 〈 0.05）; the number of mononuclear cells collected from the patients with high body mass was larger than that from the patients with low body mass （P 〈 0.05） , while the number of CD34^＋ was similar in unit body ④Adverse events and side effect： bone pain ,myalgia ,fatigue , headace , poor appetite, nausea and emesis, low-grade fever and the numbness of the lip, face and limbs during the collection. Symptoms disappeared 2 to 4 days after withdrawal. CONCLUSION： When the transplantation of autologus peripheral blood stem cells is performed in patients with lower limb ischemic angionosis, mobilization with G-CSF is effective for mononuclear cell and CD34^＋. Most of the patients can tolerate the mobilization, but certain dose of decoagulant should be used to prevent adverse reaction.