小續(xù)命湯含藥血清對(duì)氧糖剝奪模型大鼠星形膠質(zhì)細(xì)胞的保護(hù)作用研究

向軍 徐莉莉 楊峰 朱雯 蔡定芳

摘 要 目的：探討小續(xù)命湯含藥血清對(duì)氧糖剝奪（OGD）模型大鼠星形膠質(zhì)細(xì)胞的保護(hù)作用及其機(jī)制。方法：將體外培養(yǎng)的大鼠星形膠質(zhì)細(xì)胞隨機(jī)分為對(duì)照組、模型組和小續(xù)命湯含藥血清低、中、高濃度組，其中對(duì)照組細(xì)胞不作任何處理，模型組和小續(xù)命湯含藥血清低、中、高濃度組細(xì)胞經(jīng)OGD 2.5 h后分別在0（即模型組不加藥）、2.5%、5%、10%的小續(xù)命湯含藥血清中復(fù)氧0、3、6、12 h，采用比色法檢測細(xì)胞中乳酸脫氫酶（LDH）含量。取OGD后復(fù)氧12 h時(shí)的對(duì)照組、模型組和小續(xù)命湯含藥血清高濃度組大鼠細(xì)胞，采用熒光探針法檢測其活性氧（ROS）水平，采用免疫熒光雙染法檢測其錳超氧化物岐化酶（MnSOD）水平。結(jié)果：對(duì)照組細(xì)胞中LDH含量始終保持在較低水平;模型組細(xì)胞分別在OGD后復(fù)氧0～12 h時(shí)，其LDH含量從（110.99±17.06） U/L逐漸上升至（436.64±55.29） U/L，均顯著高于對(duì)照組（P<0.05）;與OGD后復(fù)氧相同時(shí)間點(diǎn)的模型組比較，小續(xù)命湯含藥血清低、中、高濃度組細(xì)胞中LDH含量均不同程度地降低，并表現(xiàn)出時(shí)間和劑量依賴趨勢，其中小續(xù)命湯含藥血清各濃度組細(xì)胞在OGD后復(fù)氧6、12 h時(shí)其LDH含量均較模型組顯著降低（P<0.05）。OGD后復(fù)氧12 h時(shí)，模型組細(xì)胞中ROS水平顯著高于對(duì)照組，MnSOD水平顯著低于對(duì)照組（P<0.05）;小續(xù)命湯含藥血清高濃度組細(xì)胞中ROS水平顯著低于模型組，MnSOD水平顯著高于模型組（P<0.05）。結(jié)論：小續(xù)命湯含藥血清可能通過上調(diào)MnSOD的水平，清除過量的ROS，從而減輕OGD導(dǎo)致的大鼠星形膠質(zhì)細(xì)胞損傷，發(fā)揮其保護(hù)作用。

關(guān)鍵詞 小續(xù)命湯;含藥血清;星形膠質(zhì)細(xì)胞;氧糖剝奪;保護(hù)作用;機(jī)制

ABSTRACT ? OBJECTIVE： To investigate the protective effects of drug-contained serum of Xiaoxuming ?decoction （XXM）on astrocyte of oxygen and glucose deprivation model rats， and to explore its mechanisms. METHODS： The astrocytes of rats were randomly divided into control group， model group and XXM low-dose， middle-dose， high-dose groups. The cells in the control group were not treated; after 2.5 h of OGD， model group and XXM low-dose， middle-dose， high-dose groups were reoxygenated for 0， 3， 6， 12 h in 0 （i.e. the model group was not added with drugs）， 2.5%， 5%， 10% of XXM， respectively. The content of lactate dehydrogenase （LDH） was detected by colorimetry. The reactive oxygen species （ROS） level was detected by fluorescence probe method， and the expression of Manganese superoxide dismutase （MnSOD） was determined by immunofluorescence double staining method in control group，model group and XXM high-dose group after 12 h of reoxygenation following OGD. RESULTS： The content of LDH in the control group was always kept at a low level; LDH content in the model group gradually increased from （110.99±17.06） U/L to （436.64±55.29） U/L after 0-12 h of reoxygenation following OGD， which was significantly higher than that in the control group （P<0.05）. Compared with model group at the same time point after reoxygenation following OGD， the contents of LDH in the cells of XXM low-dose， medium-dose and high-dose groups were decreased to different extents， and showed a time-and dose-dependent trend. The contents of LDH in XXM groups at 6 and 12 h after reoxygenation following OGD were significantly lower than that of the model group （P<0.05）. At 12 h after reoxygenation following OGD， the levels of ROS in model group were significantly higher than control group， while the level of MnSOD was significantly lower than control group （P<0.05）. The level of ROS in XXM high-dose group was significantly lower than model group， while the level of MnSOD was significantly higher than model group （P<0.05）. CONCLUSIONS： XXM can protect astrocyte by up-regulating levels of MnSOD， scavenging excessive oxygen free radicals， to relieve the OGD induced astrocytic injury， with protective effect.