中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
12期
2668-2670
,共3页
刘燊%冯世庆%周先虎%宁广智%张超
劉燊%馮世慶%週先虎%寧廣智%張超
류신%풍세경%주선호%저엄지%장초
脑源性神经生长因子前体%少突胶质细胞前体细胞%脊髓损伤
腦源性神經生長因子前體%少突膠質細胞前體細胞%脊髓損傷
뇌원성신경생장인자전체%소돌효질세포전체세포%척수손상
Precursor of brain-derived neurotrophic factor%Oligodenerocyte precursor cells%Spinal cord injnury
目的 观察脑源性神经生长因子前体(proBDNF)在少突胶质细胞前体细胞(OPCs)的表达及其对OPCs存活及生长的调控作用.方法 取新生C57BL/6小鼠大脑皮层分离少突胶质细胞前体细胞,体外原代培养.应用Western blot法观察proBDNF及其受体p75NTR和sortilin在OPCs内的表达水平;设置100 μg/L胎牛血清白蛋白(BSA)对照组和proBDNF梯度浓度组(1、10、100 μg/L),每组6孔细胞.应用结晶紫及β-tublin染色,观察OPCs细胞形态,评估proBDNF对OPCs生长的影响.结果 Western blot结果显示OPCs细胞内源性表达proBDNF及其受体p75 NTR和sortilin;与BSA对照组比较,proBDNF治疗组OPCs的数量显著减少[(537.67±34.93)个比(440.67±9.29)、(390.67±9.45)、(337.25±7.27)个],差异有统计学意义(P＜0.01);同时proBDNF可以抑制OPCs的生长,与BSA对照组比较,proBDNF治疗组OPCs的长突起减少,照片显示细胞平均面积减小显著[(2.44±0.71) μm2比(1.65±0.12)、(1.30±0.11)、(0.95±0.08)μm2],差异均有统计学意义(1 μg/L proBDNF组P＜0.05,其他组P＜0.01).结论 OPCs内源性表达proBDNF及其受体.proBDNF对OPCs的存活及生长具有抑制作用,该作用可能通过p75 NTR-sortilin通路介导.
目的 觀察腦源性神經生長因子前體(proBDNF)在少突膠質細胞前體細胞(OPCs)的錶達及其對OPCs存活及生長的調控作用.方法 取新生C57BL/6小鼠大腦皮層分離少突膠質細胞前體細胞,體外原代培養.應用Western blot法觀察proBDNF及其受體p75NTR和sortilin在OPCs內的錶達水平;設置100 μg/L胎牛血清白蛋白(BSA)對照組和proBDNF梯度濃度組(1、10、100 μg/L),每組6孔細胞.應用結晶紫及β-tublin染色,觀察OPCs細胞形態,評估proBDNF對OPCs生長的影響.結果 Western blot結果顯示OPCs細胞內源性錶達proBDNF及其受體p75 NTR和sortilin;與BSA對照組比較,proBDNF治療組OPCs的數量顯著減少[(537.67±34.93)箇比(440.67±9.29)、(390.67±9.45)、(337.25±7.27)箇],差異有統計學意義(P＜0.01);同時proBDNF可以抑製OPCs的生長,與BSA對照組比較,proBDNF治療組OPCs的長突起減少,照片顯示細胞平均麵積減小顯著[(2.44±0.71) μm2比(1.65±0.12)、(1.30±0.11)、(0.95±0.08)μm2],差異均有統計學意義(1 μg/L proBDNF組P＜0.05,其他組P＜0.01).結論 OPCs內源性錶達proBDNF及其受體.proBDNF對OPCs的存活及生長具有抑製作用,該作用可能通過p75 NTR-sortilin通路介導.
목적 관찰뇌원성신경생장인자전체(proBDNF)재소돌효질세포전체세포(OPCs)적표체급기대OPCs존활급생장적조공작용.방법 취신생C57BL/6소서대뇌피층분리소돌효질세포전체세포,체외원대배양.응용Western blot법관찰proBDNF급기수체p75NTR화sortilin재OPCs내적표체수평;설치100 μg/L태우혈청백단백(BSA)대조조화proBDNF제도농도조(1、10、100 μg/L),매조6공세포.응용결정자급β-tublin염색,관찰OPCs세포형태,평고proBDNF대OPCs생장적영향.결과 Western blot결과현시OPCs세포내원성표체proBDNF급기수체p75 NTR화sortilin;여BSA대조조비교,proBDNF치료조OPCs적수량현저감소[(537.67±34.93)개비(440.67±9.29)、(390.67±9.45)、(337.25±7.27)개],차이유통계학의의(P＜0.01);동시proBDNF가이억제OPCs적생장,여BSA대조조비교,proBDNF치료조OPCs적장돌기감소,조편현시세포평균면적감소현저[(2.44±0.71) μm2비(1.65±0.12)、(1.30±0.11)、(0.95±0.08)μm2],차이균유통계학의의(1 μg/L proBDNF조P＜0.05,기타조P＜0.01).결론 OPCs내원성표체proBDNF급기수체.proBDNF대OPCs적존활급생장구유억제작용,해작용가능통과p75 NTR-sortilin통로개도.
Objective To investigate whether the precursor of Brain-drived neurotrophic factor (proBDNF) can be expressed on oligodenerocyte precursor cells (OPCs) and the effects of proBDNF on the survival and growth of OPCs in the acute period of spinal cord injury (SCI).Methods Primary cuhure of OPCs from neonatal C57BL/6 mice was applied for the studies.Western blotting was used to investigate the expression levels of proBDNF and its receptors p75NTR and sortilin in OPCs.100 μg/L bovine serum albumin (BSA) was used as a control group and gradient concentrations of proBDNF (1,10,100 μg/L) were used to treat the cells.Each group had 6 wells of cultured cells.After the treatment,crystal violet staining and β-tublin immunofluorescence staining were used to observe the size and growth of cells.Results Western blotting showed that proBDNF,p75NTR and sortilin were endogeneously expressed on OPCs.As compared with the BSA control,proBDNF inhibited survival of OPCs (537.67 ± 34.93 vs.440.67 ± 9.29,390.67 ± 9.45 and 337.25 ± 7.27) and the difference was statistically significant (P ＜ 0.001).Meanwhile,proBDNF also inhibited the OPCs growth as compared with BSA group.The OPCs in proBDNF treated groups had less long projections and smaller size [(2.44 ± 0.71) vs.(1.65 ± 0.10),(1.30 ± 0.11) and (0.95 ± 0.08) μm2].The difference was statistically significant (P ＜ 0.05 in 1 μg/L proBDNF group,and P ＜ 0.01 in other groups).Conclusion OPCs express proBDNF and its receptors endogenously.proBDNF can inhibit the survival and growth of OPCs and the inhibitory effects may be induced by p75NTR-sortilin pathway.
