欖香烯誘導免疫原性細胞死亡及SAP62的作用

鄒治銘，劉金葉，郭連英，范琳琳，施廣霞(大連醫(yī)科大學病理生理教研室，遼寧大連116044)

欖香烯誘導免疫原性細胞死亡及SAP62的作用

鄒治銘，劉金葉，郭連英，范琳琳，施廣霞△
(大連醫(yī)科大學病理生理教研室，遼寧大連116044)

目的:探討免疫原性細胞死亡和剪接體相關蛋白62(splicesome-associated protein 62，SAP62)增強欖香烯復合瘤苗免疫原性的作用和機制。方法: (1)取30只健康BALB/c(SPF)小鼠分為瘤苗免疫組、生理鹽水對照組和聯(lián)用細胞因子組，所有小鼠在末次免疫后10 d(記為攻擊后第0天)于左腋下接種Hca-F細胞。觀察腫瘤生長情況和生存時間。(2) MTT法測細胞增殖。(3) Western blot法、間接免疫熒光和流式細胞術檢測Hca-F細胞中SAP62的變化。結果: (1) EC-TCV單用或與細胞因子聯(lián)合都有明顯的免疫保護效應。(2)欖香烯、絲裂霉素C單用或聯(lián)合均能抑制Hca-F細胞增殖(P＜0. 05)。(3)欖香烯和絲裂霉素單獨或聯(lián)合處理的細胞SAP62表達下降(P＜0. 05)。結論:欖香烯復合瘤苗細胞中觀察到的細胞死亡具有免疫原性細胞死亡的特征，SAP62表達下降，藉此影響mRNA成熟和選擇性剪接異常而發(fā)揮作用。

Sixty adult male rats were randomly divided into sham exposure group and 50 Hz/100 μT ELF-EMF exposure group.After 24-week exposure，open field test，elevated plus maze，tail suspension test，forced swim test，the Morris water maze and fear conditioning showed that exposure to ELF-EMF did not induce any anxiety-like or depression-like behaviors.Moreover，ELF-EMF exposure did not affect the morphology and histology of the brain.

AIM: To address whether inflammation mediated by NF-κB activation is involved in the prevalence of anxiety-like behavior in hypoxic pulmonary artery hypertension (HPAH) rats and to investigate the underlying mechanism.METHODS: The rats were placed in a normobaric hypoxic chamber with a fraction of inspired oxygen (FiO2) of～10%，23 h/d，continues for 2 weeks.Mean pulmonary arterial pressure (mPAP) and mean right ventricular pressure (mRVP) were measured by pulmonary artery catheterization.Anxietylike behavior was tested by elevated plus maze and open field.Inflammatory response，nucleus translocation of NF-κB，and the pathway were also examined.RESULTS: The rats in hypoxia group developed a pronounced HPAH and anxiogenic symposium，and anxiety-like behavior was positively correlated with mPAP，mRVP，and right ventricular hypertrophy (RVH)，confirming an increase in anxiety-like behavior accompanied in HPAH rats.Mechanistically，NF-κB inhibitor treatment significantly diminished mPAP，mRVP，RVH，anxiety-like behavior，and inflammatory response induced in HPAH rats，suggesting that anxiolytic by an inhibition of inflammation was through NF-κB inactivation.In addition，HPAH rats exhibited an increase in Iba-1，nucleus translocation of p65 and p50，and iNOS expression in hippocampus，indicating that activation of NF-κB in hippocampus triggered the proinflammatory response of microglia cells through iNOS pathway，which was involved in the progression of HPAH and induction of anxiety-like behavior.CONCLUSION: The crosstalk between the pathogenesis of HPAH and induction of anxiety-like behavior in rats is through inflammation by NF-κB activation with iNOS pathway in microglia ce lls in hippocampus.

*［Foundation item］Supported by Natural Science Foundation of Zhejiang Province of China (No.Y2091033)，F(xiàn)oundation of Zhejiang Educational Committee (No.Y201430622) and Scientific Research Foundation for the Returned Overseas Chinese Scholars，State Education Ministry

△Corresponding author E-mail: fxb@wmu.edu.cn

The present study was undertaken to investigate the role of p38 MAPK on the upregulation of GLT-1 induced by sulbactam during the mimic ischemic neuronal protection by neuron-astrocyte co-culture and astrocyte culture of neonatal rats.Following with 48 h sulbactam incubation or not，the culture was under 2 h oxygen glucose deprivation (OGD)，and then the medium was replaced by the basal medium.The profiles of neuronal death and survival were observed at 24 h after OGD by Hoechst/propidium iodide staining and MTT methods，respectively.The GLT-1 and p-p38 MAPK levels were detected by Western blotting and immunocytochemistry in astrocyte culture.The results showed that 2 h OGD induced significant neuronal death and sulbactam dose-dependently decreased the cell death rate and increased the cell viability against OGD，in which 125 μmol/L sulbactam had the best drug efficacy，indicating that sulbactam protects the hippocampal neurons against OGD.Sulbactam incubation dose-dependently upregulated GLT-1 expression in normal and OGD state.In normal astrocyte culture，the GLT-1 upregulation by sulbactam started at 12 h，and reached the peak at 48 h.The GLT-1 expression was downregulated at 12 h and 24 h after OGD compared with control group.Sulbactam pretreatment dose-dependently upregulated GLT-1 expression，in which 125 μmol/L sulbactam also had the best effect，indicating that sulbactam protects the neurons against OGD due to the GLT-1 upregulation in astrocyte.Although p38 expression was not changed by different doses of sulbactam，p-p38 MAPK level in normal astrocytes was upregulated dose-dependently by sulbactam.The p-p38 MAPK level was obviously earlier than that of GLT-1，in which the p-p38 started at 1 h，reached the peak at 3 h and 6 h and fell back to the baseline at 24 h，suggesting that p38 MAPK might be an upstream signal mechanism for GLT-1 upregulation by sulbactam.To prove this possibility，we further showed that the specific inhibitor of p-p38 MAPK，SB203580，dose-dependently inhibited the GLT-1 upregulation by sulbactam in astrocyte culture either in normal or OGD state.Furthermore，the p-p38 MAPK inhibitor dose-dependently overturned the protection of sulbactam on co-cultured neurons.In conclusion，sulbactam protects cerebral neurons against ischemia by upregulating GLT-1 expression via p38 MAPK signal pathway in ast rocytes.

△Corresponding author

Effects of extremely low-frequency electromagnetic fields (100 μT) on behaviors in rats

CHEN Chen
(Tongji Hospital，Huazhong University of Science＆Technology，Wuhan 430030，China.E-mail: aids_0_111@126.com)

Blunted inflammation mediated by NF-κB activation in hippocampus alleviates anxiety-like behavior in hypoxic pulmonary artery hypertension rats*

LI Yang，WANG Wei，DING Lu，ZHENG Qing-qing，GUO Jin-bin，F(xiàn)AN Xiao-fang，GONG Yong-sheng，F(xiàn)AN Jun-ming△
(Institute of Hypoxia Medicine，Wenzhou Medical University，Wenzhou 325035，China)

Sulbactam protects hippocampal neurons against oxygen-glucose deprivation by upregulation of GLT-1 via p38 MAPK signal pathway in astrocytes

QI Jie1，ZHANG Jing-ge1，ZHANG Min1，HU Yu-yan1，LI Wen-bin1，2△
(1Department of Pathophysiology，Hebei Medical University，Shijiazhuang 050017，China;2Aging and Cognition Neuroscience Laboratory of Hebei Province，Shijiazhuang 050031，China)

△E-mail: shiguangxia@163.com