中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2011年
6期
333-336
,共4页
李建生%刘敬霞%刘轲%王丁超%任伟宏%张新峰%田玉收
李建生%劉敬霞%劉軻%王丁超%任偉宏%張新峰%田玉收
리건생%류경하%류가%왕정초%임위굉%장신봉%전옥수
脑缺血%粒细胞集落刺激因子%骨髓干细胞%CD34+
腦缺血%粒細胞集落刺激因子%骨髓榦細胞%CD34+
뇌결혈%립세포집락자격인자%골수간세포%CD34+
Cerebral ischemia%Granulocyte colony stimulating factor%Bone marrow stem cell%CD34+
目的 研究人重组粒细胞集落刺激因子(rG-CSF)对骨髓干细胞(BMSCs)在脑缺血大鼠血液和脑组织中的分布变化及抗脑缺血损伤的影响.方法 将106只SD大鼠按随机数字表法分为假手术组(10只)、模型组(48只)、rG-CSF组(48只),后两组再分为术后2、3、7、14 d亚组,每个亚组12只.用改良线栓法制备大鼠局灶性脑缺血模型.rG-CSF组于术前3 d和术后2 d皮下注射rG-CSF 10μg/kg,假手术组和模型组给予等量生理盐水,均每日1次.术后各时间点进行神经功能评分;取腹主动脉血,测定外周血白细胞计数(WBC)及CD34+细胞计数;观察脑组织病理改变;并用免疫组化法测定脑组织CD34+细胞表达.结果 ①制模后2 d大鼠神经功能评分即显著降低,随后逐渐升高;rG-CSF组术后7 d和14 d神经功能评分(分)较模型组显著增高(7 d:11.86±0.69比10.53±0.76,14 d:13.38±0.52比12.38±0.52,均P<0.01).②制模后2 dSb周血WBC和CD34+细胞计数即显著增加,3 d达峰值,7 d和14 d降低;除14 d CD34+细胞计数外,rG-CSF组其余时间点WBC和CD34+细胞计数均较模型组明显增加[WBC(×109/L)2 d:11.75±1.76比8.07±1.27,3 d:13.07±1.70比10.88±1.78,7 d:8.63±1.36比5.58±1.57,14 d:6.98±0.98比4.87±0.92;CD34+细胞计数(个/μl)2 d:8.83±2.14比3.17±0.75,3 d:13.50±1.87比5.00±1.55,7 d:5.33±1.21比2.33±1.21,P<0.05或P<0.013.③制模后2 d大鼠脑组织CD34+细胞表达即明显增强,7 d达峰值,14 d降低;rG-CSF组各时间点CD34+表达[吸光度(A)值]均较模型组显著增加(2 d:43.21±4.41比22.04±2.95,3 d:45.79±1.76比25.69±2.44,7 d:52.09±2.86比33.04±2.62,14 d:29.73±1.99比16.91±2.95,均P<0.01).④rG-CSF组脑组织病理损伤较模型组减轻,以14 d改善明显.结论 脑缺血可引起BMSCs进入外周血及向脑组织归巢,其在外周血和脑组织的变化呈先增多再减少的特点,分别于缺血后3 d和7 d达峰值;rG-CSF可使进入外周血和脑组织的BMSCs明显增加.BMSCs动员对脑缺血损伤的保护作用明显,且随动员后时间的延长呈增强趋势.
目的 研究人重組粒細胞集落刺激因子(rG-CSF)對骨髓榦細胞(BMSCs)在腦缺血大鼠血液和腦組織中的分佈變化及抗腦缺血損傷的影響.方法 將106隻SD大鼠按隨機數字錶法分為假手術組(10隻)、模型組(48隻)、rG-CSF組(48隻),後兩組再分為術後2、3、7、14 d亞組,每箇亞組12隻.用改良線栓法製備大鼠跼竈性腦缺血模型.rG-CSF組于術前3 d和術後2 d皮下註射rG-CSF 10μg/kg,假手術組和模型組給予等量生理鹽水,均每日1次.術後各時間點進行神經功能評分;取腹主動脈血,測定外週血白細胞計數(WBC)及CD34+細胞計數;觀察腦組織病理改變;併用免疫組化法測定腦組織CD34+細胞錶達.結果 ①製模後2 d大鼠神經功能評分即顯著降低,隨後逐漸升高;rG-CSF組術後7 d和14 d神經功能評分(分)較模型組顯著增高(7 d:11.86±0.69比10.53±0.76,14 d:13.38±0.52比12.38±0.52,均P<0.01).②製模後2 dSb週血WBC和CD34+細胞計數即顯著增加,3 d達峰值,7 d和14 d降低;除14 d CD34+細胞計數外,rG-CSF組其餘時間點WBC和CD34+細胞計數均較模型組明顯增加[WBC(×109/L)2 d:11.75±1.76比8.07±1.27,3 d:13.07±1.70比10.88±1.78,7 d:8.63±1.36比5.58±1.57,14 d:6.98±0.98比4.87±0.92;CD34+細胞計數(箇/μl)2 d:8.83±2.14比3.17±0.75,3 d:13.50±1.87比5.00±1.55,7 d:5.33±1.21比2.33±1.21,P<0.05或P<0.013.③製模後2 d大鼠腦組織CD34+細胞錶達即明顯增彊,7 d達峰值,14 d降低;rG-CSF組各時間點CD34+錶達[吸光度(A)值]均較模型組顯著增加(2 d:43.21±4.41比22.04±2.95,3 d:45.79±1.76比25.69±2.44,7 d:52.09±2.86比33.04±2.62,14 d:29.73±1.99比16.91±2.95,均P<0.01).④rG-CSF組腦組織病理損傷較模型組減輕,以14 d改善明顯.結論 腦缺血可引起BMSCs進入外週血及嚮腦組織歸巢,其在外週血和腦組織的變化呈先增多再減少的特點,分彆于缺血後3 d和7 d達峰值;rG-CSF可使進入外週血和腦組織的BMSCs明顯增加.BMSCs動員對腦缺血損傷的保護作用明顯,且隨動員後時間的延長呈增彊趨勢.
목적 연구인중조립세포집락자격인자(rG-CSF)대골수간세포(BMSCs)재뇌결혈대서혈액화뇌조직중적분포변화급항뇌결혈손상적영향.방법 장106지SD대서안수궤수자표법분위가수술조(10지)、모형조(48지)、rG-CSF조(48지),후량조재분위술후2、3、7、14 d아조,매개아조12지.용개량선전법제비대서국조성뇌결혈모형.rG-CSF조우술전3 d화술후2 d피하주사rG-CSF 10μg/kg,가수술조화모형조급여등량생리염수,균매일1차.술후각시간점진행신경공능평분;취복주동맥혈,측정외주혈백세포계수(WBC)급CD34+세포계수;관찰뇌조직병리개변;병용면역조화법측정뇌조직CD34+세포표체.결과 ①제모후2 d대서신경공능평분즉현저강저,수후축점승고;rG-CSF조술후7 d화14 d신경공능평분(분)교모형조현저증고(7 d:11.86±0.69비10.53±0.76,14 d:13.38±0.52비12.38±0.52,균P<0.01).②제모후2 dSb주혈WBC화CD34+세포계수즉현저증가,3 d체봉치,7 d화14 d강저;제14 d CD34+세포계수외,rG-CSF조기여시간점WBC화CD34+세포계수균교모형조명현증가[WBC(×109/L)2 d:11.75±1.76비8.07±1.27,3 d:13.07±1.70비10.88±1.78,7 d:8.63±1.36비5.58±1.57,14 d:6.98±0.98비4.87±0.92;CD34+세포계수(개/μl)2 d:8.83±2.14비3.17±0.75,3 d:13.50±1.87비5.00±1.55,7 d:5.33±1.21비2.33±1.21,P<0.05혹P<0.013.③제모후2 d대서뇌조직CD34+세포표체즉명현증강,7 d체봉치,14 d강저;rG-CSF조각시간점CD34+표체[흡광도(A)치]균교모형조현저증가(2 d:43.21±4.41비22.04±2.95,3 d:45.79±1.76비25.69±2.44,7 d:52.09±2.86비33.04±2.62,14 d:29.73±1.99비16.91±2.95,균P<0.01).④rG-CSF조뇌조직병리손상교모형조감경,이14 d개선명현.결론 뇌결혈가인기BMSCs진입외주혈급향뇌조직귀소,기재외주혈화뇌조직적변화정선증다재감소적특점,분별우결혈후3 d화7 d체봉치;rG-CSF가사진입외주혈화뇌조직적BMSCs명현증가.BMSCs동원대뇌결혈손상적보호작용명현,차수동원후시간적연장정증강추세.
Objective To explore the influence of recombination granulocyte colony stimulating factor (rG-CSF)on mobilization and distribution of bone marrow stem cells (BMSCs) in blood and brain tissue,and its role in protecting brain in rats with cerebral ischemia.Methods One hundred and six SpragueDawley(SD)rats were divided into sham-operated group (n=10),model group(n=48),rG-CSF group (n=48) according to the method of random digital table,and rats in model and rG-CSF groups were divided into four subgroups:i.e.2,3,7 and 14 days subgroups,with 12 rats in each subgroup.Middle cerebral artery occlusion(MCAO)model was reproduced with nylon thread.In rats of rG-CSF group rG-CSF (10 btg/kg)was administered by subcutaneous injection 3 days before and 2 days after operation respectively,once a day.Rats in sham-operated and model groups were administered with normal saline in the same volume,once a day.At the corresponding time after operation,general neural function score(GNFS)of rats was measured.Blood was collected through abdominal aorta,then white blood cell (WBC) and CD34+ cells in peripheral blood were counted.Brain pathologic changes were observed,and expression of CD34+ cells in rats brain tissue was determined by using immunohistochemical method.Results ①GNFS was lower obviously in 2-day model group compared with that in sham-operated group,and then increased gradually.At 7 days and 14 days after operation,GNFS in rG-CSF group was higher significantly than that in model group (7 days:11.86±0.69 vs.10.53±0.76,14 days:13.38±0.52 vs.12.38±0.52,both P<0.01).②WBC and CD34+ cells in peripheral blood in model group increased obviously,with the highest level appeared at 3 days and lowered at 7 days and 14 days.Increase of WBC and CD34+ cells in rats of rG-CSF group was more obvious than that of model group at each time point except CD34+ in 14 days group [WBC (×109/L)2 days:11.75±1.76 vs.8.07±1.27,3 days:13.07±1.70 vs.10.88±1.78,7 days:8.63±1.36 vs.5.58士1.57,14 days:6.98士0.98 vs.4.87士0.92;CD34'(cells/t~1)2 days:8.83±2.14 vs.3.17±0.75,3 days:13.50±1.87 vs.5.00±1.55,7 days:5.33±1.21 vs.2.33±1.21,P<0.05 or P<0.01].③Expression of CD34+ cells in the brain of rats in 2-day model group increased significantly,and the highest level appeared at 7 days and decreased at 14 days.Absorbance (A) value of CD34+ cells expression in rat brains of each rG-CSF group was more significant than that in model group(2 days:43.21±4.41 vs.22.04±2.95,3 days:45.79±1.76 vs.25.69±2.44,7 days:52.09±2.86 vs.33.04±2.62,14 days:29.73±1.99 vs.16.91±2.95,all P<0.01).④ The signs of injury to brain in pathological examination were less obvious in 14 days rG-CSF group.Conclusion BMSCs could be induced to enter peripheral blood and "home" to brain tissue after cerebral ischemia.It was showed that BMSCs increased in number at first and then decreased in peripheral blood and brain,the peak number was found on 3rd day in peripheral blood and 7th day in brain.Mobilization with rG-CSF could increase the number of BMSCs in peripheral blood and brain tissue.The effect of mobilization of BMSCs on protecting brain was significant after cerebral ischemia,and effect appeared to be more pronounced with prolongation of mobilization.
