CAJ | 학술논문

目的：观察重组人粒细胞集落刺激因子（rhG-CSF）对缺氧人神经母细胞瘤SH-SY5Y细胞及大鼠缺血脑组织中分子伴侣介导自噬（ CMA）的标志性蛋白LAMP-2a及HSC-70表达的影响，探讨rhG-CSF抑制缺血缺氧后神经元损伤的机制。方法：制备SH-SY5Y细胞缺氧模型，分别给予0、100、200μg/L的rhG-CSF孵育48 h，Western blot检测各组细胞LAMP-2a及HSC-70的表达。将54只SD大鼠随机分为3组，每组18只；假手术组仅行颈部手术不栓塞大脑中动脉，模型组及实验组制备大脑中动脉栓塞模型，实验组给予rhG-CSF（50μg· kg-1· d-1）注射5 d，假手术组及模型组给予等量生理盐水。 Western blot 检测脑缺血周边区组织LAMP-2a及HSC-70的表达。参照改良神经功能缺损评分对3组大鼠进行神经功能评价。结果：随rhG-CSF浓度的增加，缺氧SH-SY5Y细胞HSC-70及LAMP-2a的表达逐渐升高（F＝31．574、3．943，P＜0．001）。3组大鼠HSC-70及LAMP-2a的表达水平差异有统计学意义（F＝503．658、859．985，P＜0．001），且实验组及模型组大鼠HSC-70、LAMP-2a的表达水平高于假手术组，实验组高于模型组（P＜0．05）。实验组大鼠神经功能改善程度优于模型组（t＝6．322，P＜0．001）。结论：脑缺血中rhG-CSF可能通过上调CMA发挥神经元保护作用。
목적：관찰중조인립세포집락자격인자（rhG-CSF）대결양인신경모세포류SH-SY5Y세포급대서결혈뇌조직중분자반려개도자서（ CMA）적표지성단백LAMP-2a급HSC-70표체적영향，탐토rhG-CSF억제결혈결양후신경원손상적궤제。방법：제비SH-SY5Y세포결양모형，분별급여0、100、200μg/L적rhG-CSF부육48 h，Western blot검측각조세포LAMP-2a급HSC-70적표체。장54지SD대서수궤분위3조，매조18지；가수술조부행경부수술불전새대뇌중동맥，모형조급실험조제비대뇌중동맥전새모형，실험조급여rhG-CSF（50μg· kg-1· d-1）주사5 d，가수술조급모형조급여등량생리염수。 Western blot 검측뇌결혈주변구조직LAMP-2a급HSC-70적표체。삼조개량신경공능결손평분대3조대서진행신경공능평개。결과：수rhG-CSF농도적증가，결양SH-SY5Y세포HSC-70급LAMP-2a적표체축점승고（F＝31．574、3．943，P＜0．001）。3조대서HSC-70급LAMP-2a적표체수평차이유통계학의의（F＝503．658、859．985，P＜0．001），차실험조급모형조대서HSC-70、LAMP-2a적표체수평고우가수술조，실험조고우모형조（P＜0．05）。실험조대서신경공능개선정도우우모형조（t＝6．322，P＜0．001）。결론：뇌결혈중rhG-CSF가능통과상조CMA발휘신경원보호작용。
Aim:To observe the effects of recombinant human granulocyte colony stimulating factor ( rhG-CSF ) on ex-pressions of LAMP-2a and HSC-70 in hypoxia SH-SY5Y cells and hopoxia-ischemia rats so as to further discuss the protec-tion mechanism of rhG-CSF.Methods:SH-SY5Y cell hypoxia model was prepared and cells were cultured with rhG-CSF at 0, 100, 200 μg/L for 48 h, respectively.Western blot was used to detect the expressions of LAMP-2a and HSC-70.Fifty-four SD rats were allocated into 3 groups(18 in each group).The sham operation group only received surgery .The rats in model and treatment groups were made middle cerebral artery occlusion model .The rats in the treatment group were injec-ted rhG-CSF at 50 μg· kg-1 · d-1 while those in the other 2 groups were given normal saline .The expressions of LAMP-2a and HSC-70 in brain tissue were detected by Western blot .The mNSS intelligent table was used to evaluate the neural function.Results:The expressions of HSC-70 and LAMP-2a in SH-SY5Y cells increased with the concentration of rhG-CSF (F=31.574,3.943,P<0.001).In rats, the expressions of LAMP-2a and HSC-70 of the 3 groups were significantly dif-ferent (F=503.658,859.985,P<0.001); compared with sham operation group, model group and treatment group had higher expressions of LAMP-2a and HSC-70,and compared with model group , treatment group had higher expressions of LAMP-2a and HSC-70(P<0.05).The improvement of neural function of the treatment group was significantly higher than that of the model group (t=6.322, P<0.001).Conclusion:rhG-CSF may play a protective role by increasing CMA in ischemia-induced neural injury .