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AIM: To explore the effect of tanshinone ⅡA on human osteosarcoma HOS cells and the underlying mechanism.METHODS: The cell viability and the appropriate dose of tanshinone ⅡA were determined by CCK-8 assay. Colony formation assay and Transwell assay were used to investigate the proliferation and migration abilities of the HOS cells treated with tanshinone ⅡA. The apoptosis of the HOS cells was monitored by Hoechst 33258 staining, transmission electron microscopy and flow cytometry. The protein levels of apoptosis-related molecules and JNK signaling-associated proteins were determined by Western blot. Meanwhile, a JNK inhibitor was added for confirming the relationship between the pathway and apoptosis mentioned above.RESULTS: Tanshinone ⅡA inhibited both HOS cell proliferation and migration in a dose-and time-dependent manner. Exposure of the HOS cells to tanshinone ⅡA resulted in the activation of apoptosis. Tanshinone ⅡA treatment increased the protein levels of cleaved caspase-3, Bax and JNK signaling-associated proteins, and decreased the protein level of Bcl-2, which were reversed by JNK inhibitor SP600125. Moreover, the result of CCK-8 assay revealed that tanshinone ⅡA-induced cell death was alleviated by JNK inhibitor.CONCLUSION: Tanshinone ⅡA induces cell growth inhibition and the activation of apoptosis via JNK signaling pathway in human osteosarcoma HOS cells. 相似文献
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c-Jun氨基末端激酶(c-Jun N-terminal kinase, JNK)是MAPK家族(mitogen-activated protein kinases, 丝裂原活化蛋白激酶)的重要成员, 具有参与昆虫抗逆反应等多种功能。为明确JNK在棉铃虫 Helicoverpa armigera 中的表达特性及其对Bt杀虫蛋白的应激与免疫反应, 本研究通过PCR克隆得到2个棉铃虫 JNK 基因 HaJNK1 和 HaJNK2; 生物信息学分析结果显示: HaJNK1和 HaJNK2基因开放阅读框分别为1 191、1 143 bp, 分别编码397、381个氨基酸。系统进化树分析结果表明棉铃虫 HaJNK1 与黏虫 Mythimna separata 聚为一支, 亲缘关系较近, HaJNK2与 家蚕 Bombyx mori 聚为一支, 同源性较高。利用实时荧光定量PCR技术分析 HaJNK1 与 HaJNK2 在棉铃虫不同发育时期、不同组织中的表达量, 发现 HaJNK1 与 HaJNK2 在卵中表达量最高, 其次是雌成虫; HaJNK1 在性腺中表达量最高, 其次是唾液腺; HaJNK2 在头部表达量最高, 其次是性腺。取食Cry1Ac的4龄棉铃虫幼虫的中肠组织中, HaJNK1 与 HaJNK2 的表达量均显著升高。推测 HaJNKs 基因可能参与棉铃虫抵御Bt杀虫蛋白伤害的应激和抗逆反应。 相似文献
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【目的】 研究硒蛋白谷胱甘肽过氧化物酶4（glutathione peroxidases 4,GPX4）失活如何参与调控脂多糖（lipopolysaccharide,LPS）诱导的RAW264.7巨噬细胞炎症反应及其潜在的分子机制。【方法】 体外培养RAW264.7巨噬细胞,以DMSO为对照,使用0.1~5.0 μmol/L GPX4抑制剂FIN56处理,通过CCK-8法检测细胞活力和Western blotting检测GPX4蛋白表达水平,确定抑制剂最适浓度。将RAW264.7巨噬细胞分为4组：对照组,添加DMSO培养24 h;FIN56（GPX4抑制剂）组,添加0.5 μmol/L FIN56培养24 h;DMSO-LPS组,DMSO培养24 h后使用LPS （100 ng/mL）刺激3 h;FIN56-LPS组,FIN56培养24 h后使用LPS刺激3 h。各组细胞经培养后,利用荧光探针2',7'-二氯二氢荧光素二乙酸酯（2',7'-dichlorodi-hydrofluorescein diacetate,H2DCFDA）检测细胞内活性氧（reactive oxygen species,ROS）水平,使用试剂盒法检测细胞内丙二醛（malondialdehyde,MDA）水平,分别使用ELISA和荧光定量PCR检测促炎细胞因子白细胞介素1β（interleukin-1β,IL-1β）、白细胞介素6（interleukin-6,IL-6）、肿瘤坏死因子α（tumor necrosis factor-α,TNF-α）的分泌水平和基因表达量,使用Western blotting法检测Toll样受体4（toll like receptor 4,TLR4）信号通路相关蛋白表达水平。【结果】 与DMSO处理相比,0.5 μmol/L FIN56处理巨噬细胞24 h对细胞活性无显著影响（P>0.05）,且显著降低GPX4蛋白表达水平（P<0.05）,故选用0.5 μmol/L FIN56用于后续实验。与对照组相比,FIN56和LPS均显著增加了ROS积累（P<0.05）,而与DMSO-LPS组相比,FIN56-LPS组ROS水平显著升高（P<0.05）。与对照组相比,LPS可显著增加小鼠巨噬细胞IL-1β、IL-6和TNF-α的mRNA表达量和IL-6含量,以及c-Jun氨基末端蛋白激酶（c-Jun N-terminal protein kinase,JNK）和c-Jun磷酸化水平（P<0.05）,而FIN56可显著下调LPS诱导的IL-6的mRNA表达量和含量及JNK和c-Jun磷酸化水平（P<0.05）,但对p38蛋白（p38 MAPK,p38）、细胞外调节蛋白激酶（phosphorylate extracellular regulated protein kinases1/2,Erk1/2）蛋白表达水平无显著影响（P>0.05）。【结论】 GPX4失活可有效阻断JNK和c-Jun磷酸化,从而特异性抑制LPS诱导的巨噬细胞的炎症反应。该结果可为以GPX4为靶点研发抗炎治疗策略提供理论依据。 相似文献
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LI Xin WEI Hong-yan HU Chun-lin JING Xiao-li XIONG Yan ZHAN Hong LIAO Xiao-xing 《园艺学报》2011,27(8):1552-1556
AIM: To investigate the molecular mechanism of neuronal apoptosis by observing the changes of key proteins in SAPK/JNK and Bcl-2/Bax signal pathways after brain infarction. METHODS: The cortical infarction was induced by photochemistry, namely photothrombotic cortical injury (PCI). Thirty-six Sprague-Dawley rats were randomly divided into 2 groups: PCI group and sham-operated group. The ipsilesional cortex was harvested for histomorphometry and transmission electron microscopy 7 days after PCI. Some key proteins including p-JNK1, p-JNK2, p-c-Jun, p-ATF-2, total JNK1, total JNK2, Bcl-2 and Bax were detected by Western blotting analysis.RESULTS: The cortical infarction in rats was successfully induced by photochemistry. The apoptosis of neurons in cortex was more obvious in PCI group than that in sham-operated group 7 days after PCI. The levels of p-JNK1, p-JNK2, p-c-Jun and p-ATF-2 in PCI group were significantly higher than those in sham-operated group, whereas the ratio of Bcl-2/Bax was significantly lower(P<0.05). CONCLUSION: Apoptosis is a major contributor to neuronal loss induced by cerebral hypoxia-ischemia for a long period after cortical infarction. The process is related to some apoptotic proteins such as Bcl-2/Bax and the SAPK/JNK signal pathways activated by ischemic injury. 相似文献
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LI Ming-hang TIAN Xiao-cui AN Rui-di ZHANG Qian YANG Mei XIANG Fei WANG Yu-chun XU Lu DONG Zhi 《园艺学报》2019,35(1):112-118
AIM: To investigate the effect of all-trans retinoic acid (ATRA) on blood-brain barrier after cerebral ischemia-reperfusion (CIR) injury in rats and its possible role mechanism.METHODS: Male SD rats were randomly divided into sham group, model (CIR) group and CIR+ATRA (10, 30 and 90 mg/kg) groups. The rat model of CIR injury was established by MCAO thread occlusion method. After ischemia for 1.5 h and reperfusion for 24 h, the neurological functional behavioral score, cerebral infarction volume, brain water content and Evans blue content were determined. The activity of matrix metalloprotein-9 (MMP-9) was measured by gelatin zymography. The protein levels of claudin-5, occludin, ZO-1, JNK, p-JNK, P38, p-P38 and MMP-9 in the brain tissues were determined by Western blot.RESULTS: Compared with CIR model group, ATRA at 30 mg/kg significantly improved neurological function, and decreased cerebral infarction volume, brain water content, Evans blue content and the degradation of tight junction proteins in ischemic area (P<0.01). The activity and protein expression of MMP-9 in ischemic brain tissue were decreased (P<0.01). The phosphorylation of JNK and P38 was inhibited and the protein levels of p-JNK and p-P38 were decreased (P<0.01).CONCLUSION: ATRA reduces the damage of brain tissue and the destruction of blood-brain barrier induced by CIR in rats. The protective effect may be related to inhibiting the activation of JNK/P38 MAPK signaling pathway and MMP-9. 相似文献
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Imtiaj Hasan Shigeki Sugawara Yuki Fujii Yasuhiro Koide Daiki Terada Naoya Iimura Toshiyuki Fujiwara Keisuke G. Takahashi Nobuhiko Kojima Sultana Rajia Sarkar M. A. Kawsar Robert A. Kanaly Hideho Uchiyama Masahiro Hosono Yukiko Ogawa Hideaki Fujita Jiharu Hamako Taei Matsui Yasuhiro Ozeki 《Marine drugs》2015,13(12):7377-7389
MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface. 相似文献
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肉仔鸡卫星细胞氧化应激时MAPK信号通路 总被引：1，自引：0，他引：1
【目的】在地塞米松(DEX)诱导骨骼肌卫星细胞氧化应激的体外条件下,筛选丝裂原活化蛋白激酶(MAPKs)信号系统中起关键作用的信号通路。【方法】 将体外培养的肉仔鸡胸肌骨骼肌卫星细胞(SCs)分别进行处理：Ⅰ、对照组;Ⅱ、DEX;Ⅲ、p38 MAPK抑制剂;Ⅳ、DEX+p38 MAPK抑制剂;Ⅴ、JNK抑制剂;Ⅵ、DEX+JNK抑制剂;Ⅶ、ERK5抑制剂;Ⅷ、DEX+ERK5抑制剂;Ⅸ、ERK1/2抑制剂;Ⅹ、DEX+ERK 1/2抑制剂。除处理Ⅰ和处理Ⅱ外,其它处理首先用抑制剂预处理30 min,弃去培养基后,再按处理设置分别加入含DEX的培养基或者正常培养基继续处理24 h。处理结束后,分别测定细胞培养液中丙二醛(MDA)、活性氧自由基(ROS)、超氧化物歧化酶(SOD)和谷胱甘肽硫转酶(GST)含量或活性;同时采用RT-PCR方法检测通路蛋白及SOD与GST基因表达量。【结果】 抑制剂单独处理组MDA和ROS的产生量显著低于DEX处理(P＜0.05);DEX+ERK5抑制剂处理与DEX+ERK 1/2抑制剂处理的MDA和ROS产生量显著高于ERK 5抑制剂处理和ERK1/2抑制剂处理(P＜0.05); DEX+p38 MAPK抑制剂处理与 DEX+JNK抑制剂处理组的MDA和ROS产生量与p38 MAPK处理和JNK处理的差异不显著(P＞0.05)。抑制剂单独处理组的SOD和GST活性比DEX处理高(P＜0.01);DEX+ERK5抑制剂处理与 DEX+ERK 1/2抑制剂处理的SOD和GST活性极显著低于ERK5抑制剂处理组和ERK1/2抑制剂处理组(P＜0.01),DEX+p38 MAPK抑制剂处理与DEX+JNK抑制剂处理的SOD和GST活性与p38 MAPK抑制剂处理和 JNK抑制剂处理的差异不显著(P＞0.05)。基因表达结果表明,DEX能够抑制GST和SOD基因的表达,抑制率达37%以上。与抑制ERK5和ERK1/2通路不同,抑制p38 MAPK和JNK通路后,DEX对GST和SOD基因的表达无明显抑制作用。【结论】 在DEX诱导的肉仔鸡胸肌骨骼肌SCs产生的氧化应激过程中,p38 MAPK和JNK通路起着关键作用。 相似文献
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JNKs是丝裂原活化蛋白激酶(MAPK)家族重要成员,参与应激反应和动物体轴的构建。为了进一步了解JNK家族在动物发育中的功能,首次分离和克隆了金鱼jnk3基因,研究了jnk3在金鱼和斑马鱼组织特异性和胚胎发育特异性表达状况。金鱼jnk3基因cDNA总长1 794 bp,其中阅读框1 293 bp,编码430个氨基酸。对jnk3基因序列及其推导的氨基酸序列进行同源性分析,结果显示,jnk3基因在脊椎动物较为保守,金鱼jnk3基因核苷酸序列与斑马鱼、人的同源性分别为94.1%和80.7%,金鱼jnk3基因氨基酸序列与斑马鱼、人的同源性分别为99.7%和93.4%。jnk3基因在鱼类成体中的表达存在着显著的组织差异,在金鱼和斑马鱼脑和精巢组织的表达具专一性,此外在斑马鱼心脏中也检测到jnk3的表达。RT-PCR检测结果显示,在鱼类胚胎发育中,神经胚期开始检测到jnk3基因mRNA表达,从神经胚到心跳期胚胎,jnk3基因表达水平逐渐增高,此后直至出膜一直保持较高的表达水平。研究表明,jnk3基因可能在鱼类后期的胚胎发育和成体脑、精巢与心脏发育中具有重要作用。 相似文献
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[Objectives]To investigate the effects of losartan on cell apoptosis and the expression of caspase-3 and JNK proteins in kidney tissue in the adenine-induced re... 相似文献
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AIM: To investigate the effect of dihydroartemisinin (DHA) adjuvant treatment on enhancing the antitumor effect of 5-fluorouracil (5-FU) against gastric cancer. METHODS: The gastric cancer BGC-823 cells were divided into control group, DHA group, 5-FU group, 5-FU+DHA group and 5-FU+DHA+SIRT1 plasmid group. The viability of BGC-823 cells treated with DHA and 5-FU was measured by MTT assay. The expression of SIRT1 and NADPH oxidase, activation of caspase-9 and caspase-3, and phosphorylation of ASK1 and JNK in the BGC-823 cells treated with DHA and 5-FU were determined by Western blot. The production of ROS and the apoptosis of the BGC-823 cells treated with DHA and 5-FU were analyzed by flow cytometry. RESULTS: Dihydroartemisinin significantly inhibited the expression of SIRT1 and increased NADPH oxidase protein level (P<0.05). DHA increased the sensitivity of BGC-823 cells to 5-FU, thus decreasing the IC50 of 5-FU to the gastric cancer cells. However, transfection with SIRT1 plasmid decreased the cytotoxicity of DHA and 5-FU co-treatment to the BGC-823 cells. DHA promoted the production of ROS and phosphorylation of ASK1 and JNK induced by 5-FU in the BGC-823 cells (P<0.05). However, ROS scavenger N-acetylcysteine (NAC) or JNK specific inhibitor SP600125 inhibited the cell death and activation of caspase-9 and caspase-3 induced by DHA and 5-FU co-treatment (P<0.05). In addition, NAC significantly inhibited the phosphorylation of JNK in the BGC-823 cells co-treated with DHA and 5-FU. However, treatment with SP600125 did not influence the ROS production in the BGC-823 cells, indicating that JNK was the downstream target of ROS pathway. CONCLUSION: Combination of DHA with 5-FU induces caspase-dependent apoptosis in gastric cancer cells through the SIRT1/NADPH oxidase/ROS/JNK signaling pathway. 相似文献