CAJ | 학술논문

目的：探讨绿原酸对淀粉样β蛋白(amyloid beta-protein，Aβ)引起的小脑颗粒细胞损伤的保护作用。方法：体外培养小脑颗粒细胞和腹腔巨噬细胞，将经过10μmol/L Aβ预处理48h的巨噬细胞上清液移入神经细胞中，同时加200μmol/L的绿原酸。培养2天后，采用MTT法、western bolt和乳酸脱氢酶检测试剂盒检测细胞活性、微管相关蛋白-2(microtubule-associated protein-2，MAP-2)表达水平和乳酸脱氢酶(lactic dehydrogenase，LDH)漏出量。结果：将Aβ预处理的巨噬细胞上清液移入到单独培养的小脑颗粒细胞48h后，可使小脑颗粒细胞活性明显降低，仅为正常的28.35%，加入绿原酸后，细胞活性明显增加达到正常的75.82%(P<0.01)。而在正常培养的神经细胞+Aβ组中，小脑颗粒细胞仅有少许死亡。在加入巨噬细胞上清液的小脑颗粒细胞组中，MAP-2表达量明显下降，仅为正常培养组的31.23%，加入绿原酸后，MAP-2的表达量则显著回升(P<0.01)，达到正常的79.77%。LDH漏出量明显增加，由402nkat/L增加到3046nkat/L，加入绿原酸后，可以使LDH漏出量明显下降，只有1285nkat/L。结论：①Aβ蛋白是通过激活巨噬细胞，使其释放毒性物质，从而引起神经细胞的损伤。②绿原酸对Aβ引起的神经细胞损伤具有保护作用。
목적：탐토록원산대정분양β단백(amyloid beta-protein，Aβ)인기적소뇌과립세포손상적보호작용。방법：체외배양소뇌과립세포화복강거서세포，장경과10μmol/L Aβ예처리48h적거서세포상청액이입신경세포중，동시가200μmol/L적록원산。배양2천후，채용MTT법、western bolt화유산탈경매검측시제합검측세포활성、미관상관단백-2(microtubule-associated protein-2，MAP-2)표체수평화유산탈경매(lactic dehydrogenase，LDH)루출량。결과：장Aβ예처리적거서세포상청액이입도단독배양적소뇌과립세포48h후，가사소뇌과립세포활성명현강저，부위정상적28.35%，가입록원산후，세포활성명현증가체도정상적75.82%(P<0.01)。이재정상배양적신경세포+Aβ조중，소뇌과립세포부유소허사망。재가입거서세포상청액적소뇌과립세포조중，MAP-2표체량명현하강，부위정상배양조적31.23%，가입록원산후，MAP-2적표체량칙현저회승(P<0.01)，체도정상적79.77%。LDH루출량명현증가，유402nkat/L증가도3046nkat/L，가입록원산후，가이사LDH루출량명현하강，지유1285nkat/L。결론：①Aβ단백시통과격활거서세포，사기석방독성물질，종이인기신경세포적손상。②록원산대Aβ인기적신경세포손상구유보호작용。
AIM: To ohserve the protective effect of chlorogenic acid on injury of cerebellar granular cells induced by amyloid β protein(Aβ) . METHODS: Cerebellar granular cells and peritoneal macrophage were cultured solely. The Aβ were added into the cultured medium and the final concentration were 10 μmol/ L for 48hours. The supernatant of macrophage were put into granular cells culture system, at same time, the chlorogenic acid were added into the culture medium and made the final concentration of chlorogenic acid to 200μmol/L, and the granular cells were continued culture for 48hous,. The levels of cytoactive, microtubule-associated protein-2(MAP-2) and leakage volume of lactate dehydrogenase(LDH) were detected with MTT, immuno-histochemical method and lactate dchydrogenase test kit. RESULTS: Supernatant of macrophage activated by Aβ moved into granular cells cultured system for 48 hours. which could decrease the cell viability of granular cells significantly to 28.35%(P< 0.01). After the chlorogenic acid were added into that medium, the cell viability were increased obviously from 28.35% to 75.82%(P<0.01). But for the group of normal granular cell with Aβ(10 μmol/ L), the granular cells only had little percent dead. After the supernatant of macrophage activated by Aβ added into the granular cells 48 hours, the MAP-2 expression of granular cells were decreased significantly compared with normal cultured granular cells, as 31.23% of normal cells. At the same time, the chlorogenic acid added into the cultured medium, the MAP-2 expression were significantly picked up (P<0.01),which get to the 79.77 percent of normal culture cells. The leakage volume of LDH increased obviously from 402 nkat/L(normal cultured granular cells) to 3046 nkat/L in the supernatant of macrophage added into normal cultured granular cells. The chlorogenic acid could reduce markedly the leakage volume of lactate dehydrogenase, which only 1285nkat/L. CONCLUSION: ① Macrophagcs could be activated by amyloid heta-protein and relcase toxic suhstance and resulted in the damage to neurons. ②The chlorogenic acid could play a role in protecting the neurons against the impairments induced by amyloid heta-protein.