CAJ | 학술논문

目的评价丙泊酚后处理对脂多糖(LPS)诱导大鼠脑组织小胶质细胞炎性反应的影响.方法原代培养SD大鼠全脑小胶质细胞,以1×105个/ml密度接种于24孔培养板(lml/孔),100个培养孔,采用随机数字表法,将其分为5组(n=20),正常对照组(C组)常规培养;L组加入1 μg/ml LPS孵育24 h;P25组、P50组和P100组于1μg/ml LLPS孵育24h时加入丙泊酚,终浓度分别为25、50、100μmol/L.于丙泊酚孵育1h时收集细胞,采用RT-PCR法检测诱导型一氧化氮合酶(iNOS) mRNA、环氧化酶-2(COX-2) mRNA、肿瘤坏死因子-α(TNF-α) mRNA和白细胞介素-1β mRNA的表达水平;收集上清液,采用Griess法测定一氧化氮(NO)浓度,采用酶联免疫吸附法测定前列腺素E2(PGE2)、TNF-α和IL-1β的浓度.结果与C组比较,L组iNOS mRNA、COX-2 mRNA、TNF-α mRNA和IL-1β mRNA的表达及上清液NO、PGE2、TNF-α和IL-1β的浓度升高(P＜0.01);与L组比较,P50组和P100组iNOS mRNA、COX-2 mRNA、TNF-α mRNA和IL-1β mRNA的表达及上清液NO、PGE2、TNF-α和IL-1β的浓度降低(P＜0.05或0.01),而P25组上述指标差异无统计学意义(P＞0.05).结论 50、100μmol/L丙泊酚后处理可抑制LPS诱导大鼠脑组织小胶质细胞炎性反应.
목적 평개병박분후처리대지다당(LPS)유도대서뇌조직소효질세포염성반응적영향.방법 원대배양SD대서전뇌소효질세포,이1×105개/ml밀도접충우24공배양판(lml/공),100개배양공,채용수궤수자표법,장기분위5조(n=20),정상대조조(C조)상규배양;L조가입1 μg/ml LPS부육24 h;P25조、P50조화P100조우1μg/ml LLPS부육24h시가입병박분,종농도분별위25、50、100μmol/L.우병박분부육1h시수집세포,채용RT-PCR법검측유도형일양화담합매(iNOS) mRNA、배양화매-2(COX-2) mRNA、종류배사인자-α(TNF-α) mRNA화백세포개소-1β mRNA적표체수평;수집상청액,채용Griess법측정일양화담(NO)농도,채용매련면역흡부법측정전렬선소E2(PGE2)、TNF-α화IL-1β적농도.결과 여C조비교,L조iNOS mRNA、COX-2 mRNA、TNF-α mRNA화IL-1β mRNA적표체급상청액NO、PGE2、TNF-α화IL-1β적농도승고(P＜0.01);여L조비교,P50조화P100조iNOS mRNA、COX-2 mRNA、TNF-α mRNA화IL-1β mRNA적표체급상청액NO、PGE2、TNF-α화IL-1β적농도강저(P＜0.05혹0.01),이P25조상술지표차이무통계학의의(P＞0.05).결론 50、100μmol/L병박분후처리가억제LPS유도대서뇌조직소효질세포염성반응.
Objective To evaluate the effects of postconditioning with propofol on lipopolysaccharide (LPS)-induced inflammatory responses of microglial cells in rat brain tissues.Methods The primary cultured microglial cells in brain tissues of Sprague-Dawley rats were seeded in 24 multi-well plates at a density of 1 × 105 cells/ml,and the microglial cells of 100 wells were randomly divided into 5 groups (n =20 each) using a random number table:control group (group C),LPS group (group L),and propofol 25,50 and 100 μmol/L groups (P25,P50,P100 groups).The cells were cultured routinely in group C.LPS 1 μg/ml was added and the cells were incubated for 24 h in group L.In P25,P20,and P100 groups,when the cells were incubated for 24 h with LPS 1 μg/ml,propofol with the final concentrations of 25,50 and 100 μmol/L was added,respectively.The cells were collected at 1 h of incubation with propofol for determination of the expression of inducible nitric oxide synthase (iNOS) mRNA,cyclooxygenase-2 (COX-2) mRNA,tumor necrosis factor-α (TNF-α) mRNA and interleukin-1β (IL-1β) mRNA (by RT-PCR).The supematant was separated for determination of the concentrations of nitric oxide (NO) (by Griess method) and pmstaglandin E2 (PGE2),TNF-α and IL-1β (by ELISA).Results Compared with group C,the expression of iNOS mRNA,COX-2 mRNA,TNF-α mRNA and IL-1β mRNA was significantly up-regulated and the concentrations of NO,PGE2,TNF-α and IL-1β in the supematant were increased in group L (P ＜ 0.01).Compared with group L,the expression of iNOS mRNA,COX-2 mRNA,TNF-α mRNA and IL-1β mRNA was significantly down-regulated and the concentrations of NO,PGE2,TNF-α and IL-1β in the supernatant were decreased in P50 and P100 groups (P ＜ 0.05 or 0.01),while no significant change in the indexes mentioned above was found in P25 group (P ＞ 0.05).Conclusion Postconditioning with propofol 50 and 100 μmol/L can inhibit LPS-induced inflammatory responses of microglial cells in rat brain tissues.