丝裂素活化蛋白激酶

摘要： Objective To evaluate the effect and mechanism of rivaroxaban, an inhibitor of coagulation factor Ⅹa (FⅩa), on endotoxin-induced injury to human umbilical vein endothelial cells (HUVEC). Methods When cultured HUVEC grow to 80% fusion, they were divided into four groups according to the random number method: blank control group (DMEM medium), lipopolysaccharide (LPS) group (cells were challenged by 100 μg/L LPS for 16 hours), FⅩa+LPS group (cells were challenged by LPS for 16 hours after they were cultured with 100 nmol/L FⅩa for 24 hours), and FⅩa+RIV+LPS group (cells were challenged by LPS for 16 hours after they were cultured with 100 nmol/L FXa and 1 μmol/L rivaroxaban for 24 hours). After each group of cells were challenged with LPS, the cell activity was detected by the cell proliferation and toxicity kit (CCK-8); the cell migration ability was detected by cell scratch experiments;the abilities of cells migration were measured by scratch-wound-healing assay; the apoptosis of cells were evaluated using flow cytometry; the endothelial barrier of cells was assessed by Transwell and Evans blue; the levels of tumor necrosis factor-α(TNF-α), interleukin (IL-1β, IL-6) were detected by the enzyme linked immunosorbent assay (ELISA); the expressions of nuclear factor-κB (NF-κB) and mitogen activated protein kinase (MAPK) signaling pathway were detected by Western Blot. Results Compared with blank control group, the cell viability in LPS group was significantly decreased, and the migration ability, number of apoptotic cells, and barrier permeability of endothelial cells was significantly increased, the levels of TNF-α, IL-1β and IL-6 were significantly increased, and the expressions of phosphorylation of c-Jun N-terminal kinase (p-JNK), phosphorylation of p38MAPK (p-p38MAPK), phosphorylation of transforming growth factor kinase 1 (p-TAK1) and phosphorylation of NF-κBp65 (p-NF-κBp65) were significantly increased. It indicated that LPS could stimulate the inflammatory response of vascular endothelial cells, and had a significant impact on cell activity, apoptosis and function. There was no significant difference in above indexes between FⅩa+LPS group and LPS group, except for the level of IL-6 being higher in FⅩa+LPS group. Compared with FⅩa+LPS group, in FⅩa+RIV+LPS group, the cell activity was significantly increased (A value: 0.42±0.02 vs. 0.33±0.02), and migration ability was significantly decreased (folds: 1.78±0.17 vs. 2.24±0.20), the number of apoptotic cells was significantly decreased [(11.30±0.70)% vs. (21.03±0.19)%], and permeability of monolayers endothelial cells was significantly decreased [(149±12)% vs. (253±15)%], the levels of inflammatory cytokines were significantly decreased [IL-1β(ng/L): 163.2±20.7 vs. 477.8±20.2, IL-6 (ng/L): 69.3±0.5 vs. 238.0±24.1, TNF-α(ng/L): 117.0±13.1 vs. 196.2±4.5], the expressions of p-TAK1 and p-NF-κBp65 were significantly decreased (p-TAK1/TAK1: 0.74±0.09 vs. 1.85±0.15, p-NF-κBp65/NF-κBp65: 1.15±0.17 vs. 2.36±0.20), with statistically significant differences (all P 0.05). Conclusion Rivaroxaban can effectively relieve the inflammatory response of HUVEC stimulated by LPS, which may be related to the inhibition of NF-κB signaling pathway activation rather than MAPK signaling pathway.%目的 探讨凝血因子Ⅹa(FⅩa)抑制剂利伐沙班对内毒素诱导的人脐静脉内皮细胞(HUVEC)损伤的作用及其机制.方法 体外培养HUVEC,待细胞生长至80%融合时,按随机数字表法分为4组:空白对照组(DMEM培养基)、脂多糖(LPS)组(100 μg/L LPS培养16 h)、FⅩa+LPS组(100 nmol/L FⅩa预处理24 h后加入LPS)、FⅩa+RIV+LPS组(100 nmol/L FⅩa+1 μmol/L利伐沙班预处理24 h后加入LPS).各组细胞处理后用CCK-8细胞增殖及细胞毒性检测试剂盒检测细胞活性,用细胞划痕实验检测细胞迁移能力,用流式细胞仪检测细胞凋亡情况,用Transwell小室法及伊文思蓝检测单层内皮细胞屏障通透性,用酶联免疫吸附试验(ELISA)检测细胞肿瘤坏死因子-α(TNF-α)、白细胞介素(IL-1β、IL-6)水平,用蛋白质免疫印迹试验(Western Blot)检测细胞核转录因子- κB(NF-κB)和丝裂素活化蛋白激酶(MAPK)炎症信号通路关键蛋白的表达.结果 与空白对照组比较,LPS组细胞活性降低,细胞迁移能力增加,凋亡细胞增多,单层内皮细胞屏障通透性增加, TNF-α、IL-1β和IL-6等促炎因子分泌增加,炎症信号通路关键蛋白磷酸化c-Jun氨基末端激酶(p-JNK)、磷酸化p38MAPK(p-p38MAPK)、磷酸化转化生长因子激酶1(p-TAK1)和磷酸化NF-κBp65(p-NF-κBp65)表达增加,说明LPS可以刺激血管内皮细胞的炎症反应,从而对细胞活性、凋亡及功能有明显影响.FⅩa+LPS组除IL-6水平显著高于LPS组外,其他指标与LPS组比较差异均无统计学意义.与FⅩa+LPS组比较,FⅩa+RIV+LPS组细胞活性明显提高(A值:0.42±0.02比0.33±0.02),细胞迁移能力明显下降(倍:1.78±0.17比2.24±0.20),凋亡细胞明显减少〔(11.30±0.70)%比(21.03±0.19)%〕,单层内皮细胞通透性明显降低〔(149±12)%比(253±15)%〕,炎性因子水平明显降低〔IL-1β(ng/L):163.2±20.7比477.8±20.2,IL-6(ng/L):69.3±0.5比238.0±24.1,TNF-α(ng/L):117.0±13.1比196.2±4.5),炎症信号通路中p-TAK1和p-NF-κBp65蛋白表达水平明显降低(p-TAK1/TAK1 :0.74±0.09比1.85±0.15,p-NF-κBp65/NF-κBp65 :1.15±0.17比2.36±0.20),差异均有统计学意义(均P＜0.05);而p-JNK和p-p38MAPK蛋白表达水平差异无统计学意义(p-JNK/JNK:1.64±0.12比1.65±0.15,p-p38MAPK/p38MAPK :2.31±0.32比2.35±0.20,均P＞0.05).结论 利伐沙班可以有效缓解内毒素刺激HUVEC的炎症反应,其作用机制可能与抑制NF-κB信号通路激活而非MAPK信号通路有关.

摘要： 目的 观察硫化氢干预对小鼠心肌纤维化丝裂素活化蛋白激酶( MAPK)通路表达的影响.方法 取BALB/c 雌性小鼠腹部皮下注射异丙肾上腺素制备心肌纤维化模型,分为3组:模型组、硫化氢低和高剂量组.取健康小鼠为正常组.连续给药8 w.结果 与正常组比较,模型组小鼠体重明显降低(P<0. 05),心脏重量和心脏系数明显增高(P<0. 05),与模型组比较,硫化氢组体重明显增高(P<0. 05),心脏重量和心脏系数明显降低(P<0. 05),且呈剂量依赖性.与正常组比较,模型组ST段明显抬高(P<0. 05),硫化氢组ST段明显降低(P<0. 05).正常组小鼠心肌细胞正常,模型组心肌细胞肥大、肿胀,变性,部分溶解或坏死,肌原纤维扭曲,断裂,炎性细胞浸润.硫化氢低剂量组大部分病灶均被吸收,少量炎症细胞浸润,增生的纤维组织和肌原纤维溶解,硫化氢高剂量改善明显.与正常组比,模型组应激活化蛋白激酶(JNK)和p38磷酸化明显增高(P<0. 05),细胞外信号调节的蛋白激酶(ERK)和p90RSK磷酸化明显降低(P<0. 05),与模型组比较,硫化氢组JNK和p38磷酸化明显降低(P<0. 05),ERK和p90RSK磷酸化明显增高(P<0. 05),且呈剂量依赖性.结论 硫化氢能够抑制MAPK通路激活,即促进 MAPK通路中ERK活化和减少促凋亡通路JNK和p38的激活,减轻心肌纤维化,具有明显的心肌靶器官保护作用.

摘要： 目的 研究槲皮素对肿瘤坏死因子α(TNF-α)诱导的心肌成纤维细胞增殖及P38MAPK、高迁移率族蛋白B1(HMGB1)表达的影响.方法 通过胰酶消化法和差速贴壁法获得纯化的心肌成纤维细胞,应用四甲基偶氮唑蓝(MTT)法检测心肌成纤维细胞的增殖,采用Western blot法测定P38MAPK、HMGB1蛋白的表达.结果 槲皮素对基础状态下的心肌成纤维细胞的增殖无影响,但对TNF-α诱导的心肌成纤维细胞的增殖有抑制作用,且各组细胞的吸光度(A)值随槲皮素浓度的增加而减小(P＜0.05),各组细胞P38MAPK、HMGB1蛋白表达随槲皮素浓度的增加而减弱(P＜0.05).结论 槲皮素对TNF-α诱导的心肌成纤维细胞增殖有一定地抑制作用,其机制可能是通过P38 MAPK信号通路抑制HMGB1的表达,参与抑制心肌成纤维细胞的增殖.%Objective To study the effect of quercetin on proliferation and expression of P38MAPK and HMGB1 protein in neonatal rat cardiac fibroblasts induced by TNF-α.Methods Purified cardiac fibroblasts were obtained by trypsin digestion and differential adherence method.The proliferation of cardiac fibroblasts was detected by MTT assay.The expression of P38MAPK and HMGB1 protein was detected by Western blot.Results Quercetin had no effect on the proliferation of cardiac fibroblasts in the basal state but inhibited the proliferation of cardiac fibroblasts induced by TNF-α,and the A value of each group was increased with the increase of quereetin concentration(P＜0.05).The exspression of P38MAPK、HMGB1 were decresed with the icrease of quercetin.Conclusion Quercetin may inhibit the proliferation of cardiac fibroblasts induced by TNF-α.The mechanism may be inhibit the expression of HMGB1 through P38MAPK signaling pathway.

摘要： 【摘要】目的探讨急性呼吸窘迫综合征（ARDS）时血小板活化的信号通路。方法将30只健康sD大鼠按随机数字表法分为对照组（n=6）和模型组（n=24）。采用经尾静脉注射油酸0．25mL／kg制备ARDS模型；对照组给予等量生理盐水。模型组于制模后2、6、24、72h取腹主动脉血，分离血小板，采用蛋白质免疫印迹试验（WestemB10t）检测血小板丝裂素活化蛋白激酶（MAPKs）通路中的主要蛋白激酶c—Jun氨基末端蛋白激酶（JNK）的磷酸化（pJNK）水平；处死动物取肺组织，计算肺系数（肺质量，体质量×100％）及肺湿／干质量（W／D）比值；苏木素一伊红（HE）染色后，光镜下观察肺组织病理学变化。结果与对照组相比，ARDS模型组大鼠制模后2h血小板vJNS：水平即明显增高（灰度值：0．72±O．09比0．22±0．01），6h达峰值（灰度值：0．91±0．03比0．22±0．01），之后逐渐降低，至72h仍明显高于对照组（灰度值：0．39±0．06比0．22±0．0l，均P〈0．05）。ARDS模型组大鼠制模后2h大鼠肺系数和肺W／D比值即较对照组明显升高[分别为（1．30±0．20）％比（0．60±0．10）％、6．00±0．60比3．30±0．30]，之后随时间延长逐渐降低，但直至72h肺系数和肺W／D比值仍明显高于对照组[分别为（0．90±0．10）％比（0．60±0．10）％、4．80±0．70比3．30±0．30，均P〈0．05]。光镜下显示，对照组大鼠肺组织无明显病理学改变。模型组制模后2h即可见明显的肺泡水肿和间质水肿，炎性细胞浸润，小血管扩张、充血，肺泡内有大量蛋白渗出物；24h病变达极期；72h肺泡腔液体渗出大部分被吸收，肺泡腔缩小，肺泡间隔增厚，炎性细胞浸润减轻，纤维组织增生，有微血栓形成。结论ARDS时肺组织除发生病理学改变外，血小板也发生了活化，且其活化过程与JNK信号转导通路启动有关。

摘要： Objective To investigate the signal pathway of platelet activation in acute respiratory distress syndrome (ARDS). Methods Thirty healthy Sprague-Dawley (SD) rats were randomly divided into control group (n = 6) and model group (n = 24). The model of ARDS was reproduced by intravenous injection of oleic acid (0.25 mL/kg), and the rats in control group were injected with the same amount of normal saline. The blood of abdominal aorta was collected at 2, 6, 24, and 72 hours after model reproduction, the platelets were separated, and c-Jun N-terminal kinase phosphorylation (pJNK) levels which was one of major protein kinases in the mitogen-activated protein kinases (MAPKs) signal pathway was determined by Western Blot. The rats were sacrificed, the lung tissues were harvested, and lung coefficient (lung weight/body weight ×100%) and lung wet/dry (W/D) ratio were calculated. Pathological changes in the lung tissue were observed with hematoxylin-eosin (HE) staining in light microscope. Results Comp ared with the control group, platelet pJNK level in ARDS model group was significantly increased at 2 hours after model reproduction (gray value: 0.72±0.09 vs. 0.22±0.01), and peaked at 6 hours (gray value: 0.91±0.03 vs. 0.22±0.01), then it was decreased gradually. It was also significantly higher than that of control group till 72 hours after model reproduction (gray value: 0.39±0.06 vs. 0.22±0.01, all P < 0.05). Lung coefficient and lung W/D ratio in ARDS model group were significantly increased at 2 hours after model reproduction as compared with those of control group [(1.30±0.20)% vs. (0.60±0.10)%, 6.00±0.60 vs. 3.30±0.30], then they were decreased gradually. They were also significantly higher than those of control group till 72 hours after model reproduction [(0.90±0.10)% vs. (0.60±0.10)%, 4.80±0.70 vs. 3.30±0.30, all P < 0.05]. It was showed by light microscopy that lung tissue of rats in the control group had no significant pathological changes. At 2 hours after model reproduction in model group, clearly visible alveolar edema and interstitial edema, interstitial lung infiltration of inflammatory cells, small blood vessels dilation and congestion were found, and the re were a lot of protein exudates. The lesions of lung peaked at 24 hours. At 72 hours, absorption of most of fluid leaking in alveolar, alveolar space narrow, alveolar septum thickening, the reduction of inflammatory cells infiltration, fibrous tissue proliferation, and micro thrombosis formation were found. Conclusion In ARDS, in addition to pathological changes in the lung tissue, platelet activation occurs, and its activation process is related to the priming of JNK signal transduction pathways.%目的：探讨急性呼吸窘迫综合征（ARDS）时血小板活化的信号通路。方法将30只健康SD大鼠按随机数字表法分为对照组（n＝6）和模型组（n＝24）。采用经尾静脉注射油酸0.25mL/kg制备ARDS模型；对照组给予等量生理盐水。模型组于制模后2、6、24、72h取腹主动脉血，分离血小板，采用蛋白质免疫印迹试验（WesternBlot）检测血小板丝裂素活化蛋白激酶（MAPKs）通路中的主要蛋白激酶c-Jun氨基末端蛋白激酶（JNK）的磷酸化（pJNK）水平；处死动物取肺组织，计算肺系数（肺质量/体质量×100%）及肺湿/干质量（W/D）比值；苏木素-伊红（HE）染色后，光镜下观察肺组织病理学变化。结果与对照组相比，ARDS模型组大鼠制模后2h血小板pJNK水平即明显增高（灰度值：0.72±0.09比0.22±0.01），6h达峰值（灰度值：0.91±0.03比0.22±0.01），之后逐渐降低，至72h仍明显高于对照组（灰度值：0.39±0.06比0.22±0.01，均P＜0.05）。ARDS模型组大鼠制模后2h大鼠肺系数和肺W/D比值即较对照组明显升高〔分别为（1.30±0.20）%比（0.60±0.10）%、6.00±0.60比3.30±0.30〕，之后随时间延长逐渐降低，但直至72h肺系数和肺W/D比值仍明显高于对照组〔分别为（0.90±0.10）%比（0.60±0.10）%、4.80±0.70比3.30±0.30，均P＜0.05〕。光镜下显示，对照组大鼠肺组织无明显病理学改变。模型组制模后2h即可见明显的肺泡水肿和间质水肿，炎性细胞浸润，小血管扩张、充血，肺泡内有大量蛋白渗出物；24h病变达极期；72h肺泡腔液体渗出大部分被吸收，肺泡腔缩小，肺泡间隔增厚，炎性细胞浸润减轻，纤维组织增生，有微血栓形成。结论 ARDS时肺组织除发生病理学改变外，血小板也发生了活化，且其活化过程与JNK信号转导通路启动有关。

摘要： 目的：研究雨生红球藻（Haematococcus Pluvialis ）对过度训练大鼠骨骼肌 MAPK 信号通道蛋白表达及抗氧化能力的影响。方法：以等量负荷有氧运动训练和大强度耐力训练大鼠为模型，50只SPF级Wistar 7周龄雄性大鼠为对象，随机分为安静对照组（C组）、有氧训练组（M 组）、过度训练组（OM 组）、过度训练＋雨生红球藻组（HOM 组）4组，每组12只（剔除不符合实验要求的大鼠）。每天灌胃（ig ）给药1次，雨生红球藻干预组剂量为0．4 mg／kg ， ig体积为5 mL／kg ，其他组ig 等体积生理盐水。42 d力竭游泳训练后，采用Western Blot法测定各组大鼠骨骼肌丝裂素活化蛋白激酶信号系统（mitogen‐activated protein kinase ，MAPK ）的蛋白表达含量，分光光度法测定骨骼肌 SOD活性及 MDA含量。结果：有氧训练使 M 组大鼠力竭游泳时间较C组显著增加（ P＜0．05）；过度训练则使 OM 组大鼠力竭游泳时间显著低于C组（ P＜0．05）和M组（ P＜0．01）；而补充雨生红球藻可使 HOM 组大鼠的力竭游泳时间显著高于OM组（P＜0．01），与C组和 M 组相比则无显著性差异（P＞0．05）。过度训练使OM 组大鼠骨骼肌超氧化物歧化酶（Superoxide Dismutase ，SOD ）活性显著低于其他各组（ P＜0．05）；而补充雨生红球藻则使 HOM 组大鼠SOD显著高于OM 组（ P＜0．05），保持在与C组和M 组接近的水平（P＞0．05）。过度训练使OM 组大鼠骨骼肌丙二醛（Malondial‐dehyde ，MDA ）含量较C组和M 组明显升高（ P＜0．01）；而补充雨生红球藻则使 HOM 组的MDA显著低于OM 组（ P＜0．01），且与C组和M 组无显著性差异（ P＞0．05）。有氧训练使M 组大鼠p38 MAPK、细胞外信号调节蛋白激酶（extracellular‐signal regulatrd protein kinase ， ERK ）信号通路蛋白表达较 C 组显著升高（ P＜0．05）；而过度训练则使 OM 组大鼠 p38 M APK、ERK蛋白表达进一步升高，与C组相比有非常显著性差异（ P＜0．01），与 M 组相比无显著性差异（P＞0．05）；补充雨生红球藻使 HOM 组大鼠 p38 MAPK、ERK蛋白表达显著低于OM 组（P＜0．05），但高于C组（P＜0．05）。结论：1）过度训练导致MAPK信号通路蛋白过度表达，氧化‐抗氧化系统失衡，氧化应激反应加剧，对机体造成损伤；2）补充雨生红球藻可以抑制 MAPK信号通路蛋白的过度表达，提高骨骼肌抗氧化酶的活性和清除自由基的能力，调节机体的抗氧化能力，预防和延缓运动疲劳的发生与发展。%To analyze the effect of Haematococcus Pluvialis on the expression of MAPK signal channel protein and antioxidant capacity in skeletal muscle of rats with overtraining .Method :To establish equivalent load aerobic training and high intensity endurance training rats as a model ,50 7‐week‐old male SPF Wistar rats were randomly divided into 4 groups ,with 12 in each group (the rats that did not meet the requirements were removed ) :control group (C Group ) ,aerobic training group (M group ) ,overtraining group (OM group ) ,overtraining and Haematococcus Pluvialis group (HOM group ) .The dose of HOM group was 0 .4 mg/kg ,the volume was 5 ml/kg .Other groups was given equal volume of saline .After 42 days swimming training ,mitogen activated protein kinase (MAPK ) signal channel protein expres‐sion and antioxidant indicators (SOD and MDA ) in skeletal muscle of rats were evaluated by western blot and spectrophotometry .Results :The exhaustive swimming time of M group was longer than C group ( P> 0 .05 ) ,and that of HOM group was significantly longer than OM group (P 0 .05);and that in OM group was significantly lower than C group and M group (P0 .05);that in OM group was significantly higher than C group and M group (P< 0 .01) ,that in HOM group was significantly lower than OM group (P<0 .01) .The expression of p38 MAPK and ERK signal channel protein in M group was higher than that in C group ( P< 0 .05 ) ,and that in M group was lower than the OM group (P< 0 .05);that in OM group was higher than C group (P< 0 .01);that in LM group was higher than that in C group ( P< 0 .05 ) ,and that in HOM group was lower than OM group ( P < 0 .05 ) .Conclusions :1 ) The overtraining leads to an excessive expression of MAPK signal channel protein ,and an imbalance of oxidation and antioxidation system ,causes oxidative stress and damage of the body .2 ) The supplementation of Haematococcus Pluvial‐isafter Murr .can inhibit the excessive expression of MAPK signal channel protein ,and activate the MAPK signal channel so as to regulate the expression of antioxidant genes ,and improve the capacity of scavenging oxygen free radical in skeletal muscle ,and regulate the antioxidant capacity of body ,and prevent and delay the occurrence and development of exercise fatigue .

摘要： Objective:To investigate the effects of puerarin on proliferation of cerebral microvascular endothelial cells (MECs)from spontaneously hypertensive rats (SHR)and its signaling pathway.Method:Cultured cerebral MECs from SHR and WKY rats were randomly divided into experimental groups as follows:(1 )WKY control:MECs from WKY rats were incubated with vehicle (20% propanediol)for 24h.(2)SHR control:incubated MECs from SHR with 20% propanediol for 24h.(3)puerarin group:incubated with puerarin (25,50,100ng/L)for 24h.(4) FBS group:incubated MECs from SHR with 10% FBS for 24h.(5)PD98059+puerarin group:pretreated MECs from SHR with PD98059 for 10 min before puerarin (100ng/L)treatment.(6)BDM+puerarin group:pretreated with 2,3-butanedione monoxide (BDM)10 min before puerarin (100ng/L)treatment.Cell proliferation was detected by [3 H]-TdR incorporation,and phosphorylation of p42/44 mitogen-actived protein kinases (p42/44 MAPKs)were detected by western blot analysis.Results:[3 H]-TdR incorporation in MECs from puerarin (50 and 100ng/L) groups showed 74.1% and 96.5% increase,respectively,compared with SHR control (P0.05).[3 H]-TdR incorporation in MECs from PD98059 and BDM groups decreased by 44.7% and 47.5%,respectively,compared with that from puerarin (100ng/L)group (P0.05).The phosphorylation of p42 MAPK in MECs from puerarin (25,50,and 100ng/L)groups increased by 25.0%,66.7%,and 75.0% ,respectively,compared with SHR control (P0.05).Conclusion:Puerarin induces prolifera-tion of cerebral MECs from SHR,and p42/44MAPKs signaling pathway might be involved in it.%目的：探讨葛根素对自发性高血压大鼠(SHR)脑微血管内皮细胞增殖的影响及其信号转导机制。方法：体外培养 SHR及正常血压(WKY)大鼠脑微血管内皮细胞,随机分为(1)WKY 对照组：以20%丙二醇孵育24h；(2) SHR对照组：同上处理；(3)葛根素组：SHR内皮细胞以不同浓度(25、50、100ng/L)葛根素分别孵育24h；(4)胎牛血清(FBS)组：SHR内皮细胞以10%FBS孵育24h；(5)PD98059+葛根素组：SHR 内皮细胞以丝裂素活化蛋白激酶(MAPKs)抑制剂 PD98059(50μmol/L)预孵育10min,再以100ng/L 葛根素孵育24h；(6)蛋白磷酸酶激动剂(BDM)+葛根素组：培养的 SHR内皮细胞以蛋白磷酸酶激动剂2,3-丁二酮肟(20mmol/L)预孵育10min,再以100ng/L葛根素孵育24h。采用[3 H]-胸腺嘧啶核糖核苷酸([3 H]-TdR)掺入法测定各组细胞增殖,采用免疫印迹法检测各组细胞 p42/44 MAPKs磷酸化水平。结果：50ng/L 和100ng/L 葛根素组 SHR 大鼠微血管内皮细胞[3 H]-TdR掺入值较 SHR对照组分别高74.1%和96.5%(均P0.05)。PD98059和BDM+葛根素组[3 H]-TdR掺入值分别较100ng/L葛根素组低44.7%和47.5%(均P0.05)。25、50和100ng/L葛根素组p42 MAPK磷酸化水平较SHR对照组分别高25.0%、66.7%和75.0%(均P0.05)。结论：葛根素能诱导 SHR脑微血管内皮细胞增殖,其细胞内信号转导可能与 p42/44MAPKs磷酸化途径有关。

摘要： Objective To explore the relationship between phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) and diabetic macroangiopathy,and the intervention effect of atorvastatin.Methods Total of 25 healthy male Wistar rats (4 weeks old) were randomly divided into experiment group (EX,n=19) and normal control group (NC,n=6) with random number table,the rats were fed with standard rodent chow diet or high-fat,high-sugar diet in the control and EX group.The rates in EX group were injected intraperitoneally with 1％ streptozotocin(STZ)(30 mg/kg) and blood glucose≥7.8 mmol/L was considered as DM.The well-estabolished DM rats model were then randomly divided into two subgroups:diabetic control group (DM group,n=7) and atorvastatin-treated diabetic group (ATR group,n=8).The rats in ATR group were given intragastric administration of atorvastatin (10 mg· kg-1· d-1) for 8 weeks,and DM group was given drinking water in the same way.Body weight was measured weekly.The expression of phosphorylated p38 MAPK,neuclear factor(NF)-κB,and monocyte chemoattractant protein-1 (MCP-1) in the thoracic aorta and blood lipids,intercellular cell adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1) were measure by immunohistochemistry.The data were analyzed with SNK-q and Pearson correlation analysis.Results Structural destructions in DM rat aorta tissue were observed by HE staining,which characterized by endothelia cells swollen and degeneration and intima thickness.Smooth muscle disorders and collagen fiber hyperplasia in tunica media were also found in DM rats when compared to NC animals.All these pathological alterations can be improved by atorvastatin treatment.The expression of p-p38MAPK,NF-κB and MCP-1 was significantly increased in DM rats when compared to those in NC group (Z=-3.466,-3.728,-3.832,P＜0.05) and all these increasing were partly reversed by atorvastatin treatment(Z=-2.308,-2.160,-2.501,P＜ 0.05).The expression of NF-κB and MCP-1 were both found to be correlated with p-p38 MAPK levels(r=0.406,0.310,both P＜0.05).Compared with NC group,the levels of serum NF-κB,ICAM-1,VCAM-1,total cholesterol(TC),triglyceride(TG),and low-density lipoprotein(LDL) were significantly increased in DM group(t=-9.200,-5.586,-7.041,-8.788,-5.247,-5.142,all P＜ 0.05),whereas the level of high-density lipoprotein(HDL) decreased in DM group (t=5.598,P＜0.05).After 8 weeks of atorvastatin treatment,the serum levels of NF-κB,ICAM-1,VCAM-1,TG,TC,LDL were all significantly decreased compared to DM group(t=3.661,3.360,2.496,4.348,3.077,3.446,all P＜0.05).Conclusions Activation of p38 MAPK pathway plays an important role in the development of diabetic macroangiopathy.Atorvastatin may ameliorate diabetic macrovascular disease through activating p38 MAPK and decreasing vascular inflammation response.%目的 探讨p38丝裂素活化蛋白激酶(p38MAPK)与糖尿病大血管病变的关系及阿托伐他汀的干预作用. 方法 选用25只4周龄雄性Wistar大鼠,数字表法随机分为正常对照组(NC组,n=6)和实验组(EX组,n=19),EX组采用高脂高糖饲料喂养联合链脲佐菌素注射建立具有2型糖尿病动脉粥样硬化特点的动物模型.将成模糖尿病大鼠(n=15)随机分为2型糖尿病空白对照组(DM组,n=7)及阿托伐他汀干预组(ATR组,n=8).ATR组给予阿托伐他汀10 mg·kg-1·d-1灌胃8周,DM组及NC组给予等量的饮用水灌胃.测定大鼠血清、细胞间黏附分子1(ICAM-1)、血管细胞黏附分子1(VCAM-1)、血脂水平,免疫组织化学法检测大鼠胸主动脉磷酸化p38MAPK(p-p38MAPK)、核因子(NF)-κB、单核细胞趋化蛋白1(MCP-1)蛋白表达水平.数据采用SNK-q检验、Pearson相关分析进行比较分析.结果 HE染色后可见DM组大鼠主动脉管壁结构层次不清,内膜增厚,内皮细胞肿大变性,中膜平滑肌细胞排列紊乱,胶原纤维增生.与NC组相比,DM组大鼠主动脉p-p38MAPK、NF-κB、MCP-1蛋白表达水平明显升高(Z=-3.466、-3.728、-3.832;均P＜0.05).ATR组大鼠主动脉中p-p38MAPK、NF-κB、MCP-1蛋白表达水平较DM组明显降低(Z=-2.308、-2.160、-2.501,均P＜0.05).相关分析显示主动脉NF-κB、MCP-1蛋白表达和p-p38MAPK蛋白表达呈显著正相关(r=0.406、0.310,均P＜0.05).与NC组相比,DM组大鼠血清NF-κB、ICAM-1、VCAM-1、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)水平明显升高(t=-9.200、-5.586、-7.041、-8.788、-5.247、-5.142,均P＜0.05),HDL水平降低(t＝5.598,P＜0.05).治疗8周后,ATR组大鼠血清中NF-κB、ICAM-1、VCAM-1、TG、TC、LDL水平均较DM组明显降低(t=3.661、3.360、2.496、4.348、3.077、3.446,均P＜0.05),HDL水平较DM组有所升高,但组间差异无统计学意义(t=-1.983,P＞0.05). 结论 p38 MAPK通路的激活在糖尿病大血管病变发生发展中起重要作用,阿托伐他汀可通过降低p38MAPK磷酸化水平,抑制炎症反应,起到大血管保护作用.

摘要： 目的:石英和苯并芘已被列为致癌物质.本文旨在研究MAPK和AP-1信号转导通路在调节石英和苯并芘诱导的恶性转化的人胚肺成纤维细胞中cyclin D1和CDK4表达以及细胞周期改变方面的差异. 方法:采用Western blot(WB)法检测石英、苯并芘诱导的恶性转化人胚肺成纤维细胞中丝裂素活化蛋白激酶(MAPK)各亚家族(ERK、p38和JNK)表达的变化情况;以及cyclinD1和CDK4(cyclin dependent kinase)表达变化情况. 结果:与空白对照组比较,石英诱导人胚肺成纤维细胞恶性转化组P-ERK、P-JNK表达均升高(P＜0.01);CDK4、P-p38表达下降(P＜0.01,P＜0.05).苯并芘诱导人胚肺成纤维细胞恶性转化组与空白对照组比较,P-ERK、P-JNK、CyclinD1表达均升高(P＜0.01,P＜0.01和P＜0.05).苯并芘诱导人胚肺成纤维细胞恶性转化组与石英诱导人胚肺成纤维细胞恶性转化组比较,P-ERK、P-JNK表达下降(P＜0.01);P-p38、CyclinD1、CDK4表达升高(P＜0.05,P＜0.05和P＜0.01).与空白对照组比较,石英诱导人胚肺成纤维细胞恶性转化组P-ERK、P-JNK表达均升高(P＜0.01);P-p38表达下降(P＜0.05).与空白对照组比较,苯并芘诱导人胚肺成纤维细胞恶性转化组P-ERK、P-JNK、CyclinD1表达均升高(P＜0.01,P＜0.01和P＜0.05);p38表达下降(P＜0.05).苯并芘诱导人胚肺成纤维细胞恶性转化组与石英诱导人胚肺成纤维细胞恶性转化组比较,P-ERK、P-JNK表达下降(P＜0.01);P-p38、CDK4、CyclinD1表达升高(P＜0.05). 结论:在石英和苯并芘诱导的恶性转化的人胚肺成纤维细胞中MAPK/AP-1通路存在差异.

摘要： 丝裂素活化蛋白激酶(mitogen-activated protein kinases, MAPK)信号级联是细胞内重要的信号转导系统之一。细胞运用这一系统将胞外刺激信号传递给胞核，参与细胞生长、发育、分化和凋亡等一系列生理病理过程。本文介绍了MAPK的特点，浅谈了MAPK信号转导通路在疾病和损伤中的作用，阐述了MAPK与神经细胞凋亡的关系。

摘要： 目的：研究前额叶皮层MAPK通路分子在吗啡成瘾记忆中的作用.方法：采用经典的条件性位置偏爱(CPP)行为学实验及蛋白印迹技术.结果：小鼠经过8d5mg/kg吗啡注射和训练之后形成明显的条件性位置偏爱,即形成环境线索记忆,吗啡处理组小鼠在伴药箱停留的时间较对照组延长(P＜0.05),其中吗啡/CPP训练组前后时间差异明显(P＜0.05).吗啡诱导小鼠CPP形成之后,前额叶皮层脑组织中ERK1磷酸化在各实验组中没有明显差异,而给予吗啡并经过CPP训练成功的小鼠脑组织中ERK2磷酸化水平显著降低(P＜0.05).结论：MAPK通路在前额叶皮层磷酸化ERK2表达降低参与了吗啡环境线索记忆的形成.

摘要： 目的：研究前额叶皮层MAPK通路分子在吗啡成瘾记忆中的作用.方法：采用经典的条件性位置偏爱(CPP)行为学实验及蛋白印迹技术.结果：小鼠经过8d5mg/kg吗啡注射和训练之后形成明显的条件性位置偏爱,即形成环境线索记忆,吗啡处理组小鼠在伴药箱停留的时间较对照组延长(P＜0.05),其中吗啡/CPP训练组前后时间差异明显(P＜0.05).吗啡诱导小鼠CPP形成之后,前额叶皮层脑组织中ERK1磷酸化在各实验组中没有明显差异,而给予吗啡并经过CPP训练成功的小鼠脑组织中ERK2磷酸化水平显著降低(P＜0.05).结论：MAPK通路在前额叶皮层磷酸化ERK2表达降低参与了吗啡环境线索记忆的形成.

摘要： 目的：研究前额叶皮层MAPK通路分子在吗啡成瘾记忆中的作用.方法：采用经典的条件性位置偏爱(CPP)行为学实验及蛋白印迹技术.结果：小鼠经过8d5mg/kg吗啡注射和训练之后形成明显的条件性位置偏爱,即形成环境线索记忆,吗啡处理组小鼠在伴药箱停留的时间较对照组延长(P＜0.05),其中吗啡/CPP训练组前后时间差异明显(P＜0.05).吗啡诱导小鼠CPP形成之后,前额叶皮层脑组织中ERK1磷酸化在各实验组中没有明显差异,而给予吗啡并经过CPP训练成功的小鼠脑组织中ERK2磷酸化水平显著降低(P＜0.05).结论：MAPK通路在前额叶皮层磷酸化ERK2表达降低参与了吗啡环境线索记忆的形成.

摘要： 目的：研究前额叶皮层MAPK通路分子在吗啡成瘾记忆中的作用.方法：采用经典的条件性位置偏爱(CPP)行为学实验及蛋白印迹技术.结果：小鼠经过8d5mg/kg吗啡注射和训练之后形成明显的条件性位置偏爱,即形成环境线索记忆,吗啡处理组小鼠在伴药箱停留的时间较对照组延长(P＜0.05),其中吗啡/CPP训练组前后时间差异明显(P＜0.05).吗啡诱导小鼠CPP形成之后,前额叶皮层脑组织中ERK1磷酸化在各实验组中没有明显差异,而给予吗啡并经过CPP训练成功的小鼠脑组织中ERK2磷酸化水平显著降低(P＜0.05).结论：MAPK通路在前额叶皮层磷酸化ERK2表达降低参与了吗啡环境线索记忆的形成.

摘要： 目的：研究前额叶皮层MAPK通路分子在吗啡成瘾记忆中的作用.方法：采用经典的条件性位置偏爱(CPP)行为学实验及蛋白印迹技术.结果：小鼠经过8d5mg/kg吗啡注射和训练之后形成明显的条件性位置偏爱,即形成环境线索记忆,吗啡处理组小鼠在伴药箱停留的时间较对照组延长(P＜0.05),其中吗啡/CPP训练组前后时间差异明显(P＜0.05).吗啡诱导小鼠CPP形成之后,前额叶皮层脑组织中ERK1磷酸化在各实验组中没有明显差异,而给予吗啡并经过CPP训练成功的小鼠脑组织中ERK2磷酸化水平显著降低(P＜0.05).结论：MAPK通路在前额叶皮层磷酸化ERK2表达降低参与了吗啡环境线索记忆的形成.

摘要： 本发明公开了一种使有丝分裂原活化蛋白激酶失活的方法。该使有丝分裂原活化蛋白激酶失活的方法，是将含有OspF蛋白和/或SpvC蛋白和/或HopAI1蛋白的编码基因的重组真核表达载体导入目的真核细胞中，使所述目的真核细胞中的有丝分裂原活化蛋白激酶失活。

摘要： 本发明公开了一种三七丝裂原活化蛋白激酶激酶基因

摘要： 作为(MK2或MAPKAP激酶－2)抑制剂的式(I)化合物或其药学上可接受的盐或前药酯，其中基团R1－R6、A和Y如说明书中所定义。

摘要： 本发明涉及一种枸杞有丝分裂原活化蛋白激酶激酶及其在增强植物耐盐碱能力方面的应用。通过提取新鲜枸杞叶片中的总RNA，利用3’RACE方法克隆枸杞中的有丝分裂原活化蛋白激酶激酶基因LmMAPKK，得到基因完整的序列为1074bp；构建了大肠杆菌表达载体pET28a-LmMAPKK，通过大肠杆菌异源表达系统，得到了39KD大小的LmMAPKK表达蛋白；并构建了双元植物表达载体pCAMBIA2300-LmMAPKK，电击法将载体转入农杆菌C58细胞，用这种细胞转化烟草，得到转基因烟草，通过测试，发现转基因烟草对盐碱、低温及病原菌侵染的抗性大大提高，对玉米、大豆、水稻、花生、向日葵、谷子、大麦、小麦、棉花等农作物；月季、洋桔梗、安祖花、蝴蝶兰等花卉；杨树、香花槐等乔木类树木转基因，发现其转基因植株耐盐碱能力大大提高。

摘要： 本发明提供了一种在人体正常肾上腺组织中表达的新的人促分裂原活化蛋白激酶相关蛋白激酶(human MAPkinase－interacting kinase 2,简称为“hMnk2”),及其编码序列,本发明还提供了该蛋白和核酸序列的制备方法和用途,以及在样品中检测hMnk2核酸序列和多肽的方法。

摘要： 本发明涉及一种具有分子(I)结构的新型的p38裂原活化蛋白激酶抑制剂，包括了它们的制备过程，包含它们的处方组成和它们在治疗上的应用。

摘要： 本发明公开了一种使有丝分裂原活化蛋白激酶失活的方法。该使有丝分裂原活化蛋白激酶失活的方法，是将含有OspF蛋白和/或SpvC蛋白和/或HopAI1蛋白的编码基因的重组真核表达载体导入目的真核细胞中，使所述目的真核细胞中的有丝分裂原活化蛋白激酶失活。

摘要： 本发明公开了一种楤木皂苷提取物，其将楤木根皮用体积百分比浓度为65～85%的乙醇回流提取，将提取液过滤、合并浓缩为相对密度1.01后，再用水饱和的正丁醇萃取，将正丁醇层过滤、合并浓缩得楤木总皂苷质量百分含量为60-80%的楤木皂苷提取物浸膏，将浸膏干燥、粉碎得楤木皂苷提取物。上述楤木皂苷提取物可用于制备单磷酸腺苷活化的蛋白激酶激动剂，包括用于胰岛素抵抗存在的多种代谢性疾病中，如糖尿病、高血压、血脂异常、肥胖等。与以往的AMPK激动剂相比，本发明的楤木皂苷具有较强的AMPK激动活性，并且不良反应少，价格低廉，容易获得。

摘要： 本发明公开了一种基于促分裂原活化蛋白激酶激酶和组蛋白去乙酰化酶双靶点的小分子抗癌药物及其制备方法，属于小分子抗癌药物技术领域。该药物的化学结构为：

摘要： 提供了在不同的环境条件下，调节植物应答、发育和产量的方法和组合物。提供了利用MAPKKK的方法。MAPKKK序列用于多种方法中，包括调节根发育、调节叶和/或枝条发育、调节非生物胁迫下的耐受性和调节产量。还提供了转化的植物、植物细胞、组织、种子和表达载体。