Investigation on the mechanism of Qiangxinhuoli prescription in the treatment of chronic heart failure based on p38-MAPK signaling pathway

Background:The aim of this study is to investigate the mechanism of action underlying the therapeutic effects of the national patent Chinese medicine compound“Qiangxinhuoli prescription(QXHLF)”on chronic heart failure(CHF).Methods:In vitro,the H_(9)C_(2) cell model was induced by ANGII,and cell proliferation and related protein expression were detected by Cell Counting Kit-8 and Western blot.In vivo,A rat model of CHF was prepared by ligation of the left anterior descending coronary artery.The effects of QXHLF on cardiac function in CHF rats were evaluated by cardiac index,hemodynamic changes,enzyme-linked immunosorbent assay,hematoxylin-eosin staining,immunohistochemistry,Western blot and RT-PCR.The expression of pro-apoptotic factors and anti-apoptotic factors,as well as TGFβ1,p-p38,TAK 1 mRNA,and protein,were detected.Results:In vitro,QXHLF has a significant inhibitory effect on the proliferation of H_(9)C_(2) cells.QXHLF can reduce the expression levels of TAK 1,TGFβ1,p-p38,Caspase3 and BAX proteins in H_(9)C_(2) cells,and increase the expression level of BCL_(2) protein.In vivo,QXHLF has the potential to increase left ventricular systolic pressure,m aximum rate of change in left ventricular pressure while decreasing left ventricular end diastolic pressure,and inhibiting the serum levels of brain natriuretic peptide.Moreover,QXHLF exhibits significant improvements in the pathological alterations of myocardial cells and fibers in CHF rats,leading to enhanced myocardial tissue morphology and notable advantages in combating myocardial fibrosis.QXHLF can reduce the levels of BAX and Caspase3 and up-regulate the expression of BCL_(2),thereby inhibiting cardiomyocyte apoptosis.Furthermore,QXHLF demonstrates inhibitory effects on the mRNA and protein expression levels of TGFβ_(1),TAK_(1),and p-p38 in the heart tissue of the CHF rat model.Conclusion:These findings indicate that QXHLF has a therapeutic effect on CHF by inhibiting the p38-MAPK signaling pathway,reducing myocardial fibrosis,preventing apoptosis,inhibiting cell proliferation,and restoring myocardial injury.

基于p38-MAPK通路探讨真武汤对心力衰竭心肌细胞结构重构-电重构的影响

目的真武汤在治疗慢性心力衰竭(Heart Failure,HF)过程中能够改善心律失常,旨在从p38-MAPK通路调控肥大心肌结构重构和电重构角度探讨真武汤治疗HF相关心律失常的作用机制。方法用AngⅡ诱导H9c2心肌细胞构建心肌肥大模型,实验共分6组:正常组(N),模型组(M)和真武汤低剂量组(D)、中剂量组(Z)、高剂量组(G)及抑制剂组(S)。正常组(N)用含有胎牛血清的高糖培养基培养,模型组(M)用含有10^(-6) mol/L浓度的血管紧张素Ⅱ(Angio-tensinⅡ,AngⅡ)培养基处理,低(D)、中(Z)、高(G)剂量组分别予2%、4%、8%体积分数的真武汤含药血清及AngⅡ培养基共同处理,抑制剂组(S)用含有p38MAPK抑制剂SB203580的培养基处理。处理48 h后,镜下观察各组心肌细胞的生长情况,通过酶联免疫吸附试验(Enzyme Linked Immunosorbent Assay,ELISA)法测心肌肥大标志物ANP的含量,用β-半乳糖苷酶染色法检测细胞衰老情况,采用Western blot法检测各组心肌细胞缝隙连接蛋白43(Connexin 43,Cx43)、p38丝裂原激活的蛋白激酶(p38 Mitogen-ctivated Protein Kinase,p38-MAPK)、基质金属蛋白酶2(Matrix Metalloproteinase 2,MMP2)、基金金属蛋白酶9(Matrix Metalloproteinase 9,MMP9)、金属蛋白酶组织抑制剂(Tissue Inhibitor of Metalloprotei-nase1,TIMP1)、p53蛋白的表达情况,采用实时荧光定量PCR(Quantitative Real-time PCR,qPCR)定量分析各组心肌细胞ANP、Cx43、p-Cx43、p38-MAPK、MMP2、MMP9、TIMP1的mRNA转录水平,免疫荧光检测Cx43蛋白的表达和分布情况。结果高剂量的真武汤能够减轻肥大心肌细胞的表面积,中低剂量无明显效果;真武汤能够降低心肌肥大标志物ANP及其mRNA的水平,且其效果与剂量呈正相关;真武汤能改善肥大细胞的衰老情况,效果与抑制剂相似;真武汤能够降低肥大心肌细胞Cx43、p38-MAPK、MMP2、MMP9、p53、p-Cx43蛋白的表达水平及Cx43、p38-MAPK、MMP2、MMP9的mRNA转录水平,且能够增加TIMP1的表达水平及其mRNA转录水平,效果与剂量有关;真武汤能够缓解肥大细胞表面Cx43的分布紊乱及减少该蛋白在细胞侧-侧移位分布。结论真武汤可能通过p38信号通路调控心肌肥大细胞的衰老和纤维化,调节细胞外基质的代谢平衡,维持心肌细胞上Cx43数量、磷酸化水平和分布定位,改善缝隙连接重构,实现对HF心肌结构重构和电重构的统一调节,达到治疗心力衰竭及相关心律失常的目的。

Jianpi Gushen Huayu decoction ameliorated diabetic nephropathy through modulating metabolites in kidney,and inhibiting TLR4/NF-κB/NLRP3 and JNK/P38 pathways

BACKGROUND Jianpi Gushen Huayu Decoction(JPGS)has been used to clinically treat diabetic nephropathy(DN)for many years.However,the protective mechanism of JPGS in treating DN remains unclear.AIM To evaluate the therapeutic effects and the possible mechanism of JPGS on DN.METHODS We first evaluated the therapeutic potential of JPGS on a DN mouse model.We then investigated the effect of JPGS on the renal metabolite levels of DN mice using non-targeted metabolomics.Furthermore,we examined the effects of JPGS on c-Jun N-terminal kinase(JNK)/P38-mediated apoptosis and the inflammatory responses mediated by toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)/NOD-like receptor family pyrin domain containing 3(NLRP3).RESULTS The ameliorative effects of JPGS on DN mice included the alleviation of renal injury and the control of inflammation and oxidative stress.Untargeted metabolomic analysis revealed that JPGS altered the metabolites of the kidneys in DN mice.A total of 51 differential metabolites were screened.Pathway analysis results indicated that nine pathways significantly changed between the control and model groups,while six pathways significantly altered between the model and JPGS groups.Pathways related to cysteine and methionine metabolism;alanine,tryptophan metabolism;aspartate and glutamate metabolism;and riboflavin metabolism were identified as the key pathways through which JPGS affects DN.Further experimental validation showed that JPGS treatment reduced the expression of TLR4/NF-κB/NLRP3 pathways and JNK/P38 pathway-mediated apoptosis related factors.CONCLUSION JPGS could markedly treat mice with streptozotocin(STZ)-induced DN,which is possibly related to the regulation of several metabolic pathways found in kidneys.Furthermore,JPGS could improve kidney inflammatory responses and ameliorate kidney injuries in DN mice via the TLR4/NF-κB/NLRP3 pathway and inhibit JNK/P38 pathwaymediated apoptosis in DN mice.

Betulinic acid protects against ovarian impairment by decreasing F-2 toxin-induced oxidative stress and inflammation associated with the downregulation of p38 expression in mice

F-2 toxin is an estrogenic mycotoxin that causes reproductive disorders in animals.Betulinic acid(BA)is a natural pentacyclic lupane-structure triterpenoid that has diverse pharmacological activities.In this study,the antioxidative and anti-inflammatory effects of BA and its underlying mechanism are explored in F-2 toxin-triggered mouse ovarian damage.We found that BA alleviated the F-2 toxin-induced ovarian impairment by stimulating follicle growth,reducing inflammatory cell infiltration,repairing damaged mitochondria and endoplasmic reticulum.Simultaneously,BA not only reversed F-2 toxin-induced reduction of follicle stimulating hormone(FSH)and luteinizing hormone(LH)levels in the serum,but also restrained the protein expression of the estrogen receptors a(ERa)and ERβ.Moreover,BA restored the balance of F-2 toxin-induced ovarian redox system disorders.Subsequently,we found that 0.25 mg/kg BA played an anti-inflammatory role in the F-2 toxin-induced ovarian impairment by decreasing interleukin-1β(IL-1β).IL-6,and tumor necrosis factor-α(TNF-α)mRNA expression,as well as inhibiting p38 protein expression.These data demonstrated that BA exerts its protective effect on F-2 toxin-induced ovarian oxidative impairment and inflammation by inhibiting p38 expression,which implies a natural product-based medicine to ameliorate F-2 toxin-caused female reproductive toxicity and provides a detoxifying method for food contaminated by mycotoxin.

IFITM3通过P38-MAPK/EMT信号轴对胃癌细胞的影响

目的 探究IFITM3在胃癌中的表达及其对胃癌细胞恶性生物学行为的影响。方法 利用数据库分析IFITM3在胃癌中的表达及分析IFITM3与胃癌患者总体生存率的相关性;在胃癌细胞SGC-7901中转染siIFITM3,利用CCK-8实验、流式细胞术和伤口愈合实验分别分析细胞增殖、凋亡及迁移。Western blot检测P38-MAPK磷酸化水平和EMT相关蛋白(E-cadherin、Snail和Vimentin)的表达。结果 IFITM3在胃癌中高表达且与胃癌患者总体生存率负相关;在SGC-7901细胞中转染si-IFITM3能有效敲低IFITM3的表达;敲低IFITM3能显著抑制细胞增殖和迁移,诱导细胞凋亡,同时也能减少P38-MAPK的磷酸化和EMT相关蛋白的表达。结论 敲低IFITM3能通过减少P38-MAPK蛋白磷酸化,抑制胃癌细胞EMT,从而减缓细胞增殖和迁移,诱导细胞凋亡。

p38-MAPK信号通路对骨骼肌再生的影响研究进展

丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路是真核细胞功能高度保守的调节因子,在细胞受到外界刺激引起的炎症反应、应激反应等生理病理过程中起着重要调控作用。P38丝裂原活化蛋白激酶(p38 mitogen-activated protein kinase,p38-MAPK)信号通路是MAPK通路的亚族之一,在细胞炎症、组织再生、免疫反应中起着重要的作用。p38-MAPK信号通路的激活在染色质重塑和关键肌生成转录因子的激活中起着关键作用,而肌卫星细胞的激活又影响着骨骼肌的再生修复。本文对p38-MAPK信号通路的调控机制及其介导骨骼肌再生的研究进展进行了综述,以期为深入了解相关机制提供理论依据,并为进一步治疗骨骼肌损伤提供一定的临床参考。

The Antidepressant Mechanism of JiaWeiWenDan Decoction Regulating p38MAPK-ERK5 Signal Transduction Pathway

Objective: To investigate the anti-depression mechanism of JiaWeiWenDan Decoction in regulating p38MAPK-ERK5 signal transduction pathway. Methods: Depression model rats were randomly divided into Blank Control Group, Model Control Group, Chinese Medicine Treatment Group, and Western Medicine Treatment Group (hereinafter referred to as Blank Group, Model Group, Chinese Medicine Group, and Western Medicine Group), with 48 rats in each group. The mice were treated with p38MAPK-ERK5 on the 7th day, 14th day and 21st day, respectively, and the mice were treated for 28 days. The key targets and cytokines in p38MAPK-ERK5 signal transduction pathway were detected. Results: Compared with the Blank Group, the expression of p38MAPKmRNA in the hippocampus of the Model Group was increased. The Chinese Medicine Group and Western Medicine Group could reduce the expression of p38MAPK mRNA (P P P P Conclusion: The anti-inflammatory effect of JiaWeiWenDan Decoction may be related to the regulation of p38MAPK-ERK5 signaling pathway. With the advance of the treatment week, the best effect was obtained when the treatment was started on the 7th day of modeling.

C5a/C5a R pathway is essential for up-regulating Sph K1 expression through p38-MAPK activation in acute liver failure

AIM To investigate the role of the complement 5a(C5a)/C5 a receptor(C5a R) pathway in the pathogenesis of acute liver failure(ALF) in a mouse model.METHODS BALB/c mice were randomly assigned to different groups, and intraperitoneal injections of lipopolysaccharide(LPS)/D-galactosamine(D-Gal N)(600 mg/kg and 10 μg/kg) were used to induce ALF. The KaplanMeier method was used for survival analysis. Serum alanine aminotransferase(ALT) levels, at different time points within a 1-wk period, were detected with a biochemistry analyzer. Pathological examination of liver tissue was performed 36 h after ALF induction. Serum complement 5(C5), C5 a, tumor necrosis factor-α(TNF-α), interleukin(IL)-1β, IL-6, high-mobility group protein B1(HMGB1) and sphingosine-1-phosphatelevels were detected by enzyme-linked immunosorbant assay. Hepatic morphological changes at 36 h after ALF induction were assessed by hematoxylin and eosin staining. Expression of C5 a R, sphingosine kinase 1(Sph K1), p38-MAPK and p-p38-MAPK in liver tissue, peripheral blood mononuclear cells(PBMCs) and peritoneal exudative macrophages(PEMs) of mice or RAW 264.7 cells was analyzed by western blotting. C5 a R m RNA levels were detected by quantitative real-time PCR.RESULTS Activation of C5 and up-regulation of C5 a R were observed in liver tissue and PBMCs of mice with ALF. Blockade of C5 a R with a C5 a R antagonist(C5a Ra C5 a Ra) significantly reduced the levels of serum ALT, inflammatory cytokines(TNF-α, IL-1β and IL-6) and HMGB1, as well as the liver tissue damage, but increased the survival rates(P < 0.01 for all). Blockade of C5 a R decreased Sph K1 expression in both liver tissue and PBMCs significantly at 0.5 h after ALF induction. C5 a Ra pretreatment significantly downregulated the phosphorylation of p38-MAPK in liver tissues of ALF mice and C5 a stimulated PEMs or RAW 264.7 cells. Moreover, inhibition of p38-MAPK activity with SB203580 reduced Sph K1 protein production significantly in PEMs after C5 a stimulation.CONCLUSION The C5a/C5 a R pathway is essential for up-regulating Sph K1 expression through p38 MAPK activation in ALF in mice, which provides a potential immunotherapeutic strategy for ALF in patients.

p38-MAPK信号通路在HHPH中的作用及三七总皂苷干预

目的:研究p38-MAPK信号通路在大鼠低O2高CO2性肺动脉高压(HHPH)发生发展中的动态变化;探讨三七总皂苷(PNS)防治低O2高CO2性肺动脉高压的机制。方法:复制慢性HHPH大鼠模型,分为正常组(N),低O2高CO23 d组(H3d)、1周组(H1w)、2周组(H2w)、4周组(H4w),及PNS治疗组(Hp),Hp组腹腔注射血塞通注射液(成分为PNS和生理盐水)。Western印迹法、免疫组织化学技术检测肺组织及肺血管P-p38、p38蛋白的表达。结果:①H1w、H2w、H4w和Hp组的WA/TA均高于N组(P均<0.05),但H3d组较N组增加不明显(P>0.05),Hp组WA/TA明显低于H4w组(P<0.05)。②肺组织P-p38蛋白在N组表达不明显,在H3d、H1w、H2w、H4w组均有高水平表达。③肺小动脉壁P-p38蛋白在N组和H3d组表达呈阴性或弱阳性,在H1w、H2w、H4w组有高水平表达,较N组差异有统计学意义(P<0.05)。④Hp组肺组织P-p38,肺小动脉壁P-p38蛋白表达较低O2高CO2组降低(P<0.05)。结论:p38-MAPK信号通路介导了大鼠HHPH的形成。PNS可减轻这一过程,其机制可能和PNS抑制P-p38表达有关。

高良姜素通过PI3K/Akt及p38-MAPK信号通路增强胃癌SGC-7901细胞对阿帕替尼的敏感性

目的探讨高良姜素对阿帕替尼抗胃癌作用的影响及可能机制。方法体外培养胃癌SGC-7901细胞,3-( 4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(M TT)法筛选出高良姜素和阿帕替尼抑制胃癌细胞增殖的合适浓度。将胃癌细胞分为4组:空白对照组、高良姜素组、阿帕替尼组和高良姜素+阿帕替尼组。MTT法检测细胞增殖活力;流式细胞术检测细胞凋亡率;细胞划痕实验和Transwell小室实验分别检测细胞迁移和侵袭能力;Westernblot检测细胞凋亡及PI3K/Akt及p38-MAPK信号通路相关蛋白表达。结果阿帕替尼和高良姜素均显著抑制胃癌SGC-7901细胞的增殖,并呈浓度依赖性(P<0 .05), 20mg/L阿帕替尼和300mg/L高良姜素是最佳的干预浓度。与20mg/L阿帕替尼组相比,高良姜素+阿帕替尼组可以明显抑制胃癌SGC-7901细胞的增殖、迁移和侵袭(P<0.05)。流式细胞术结果显示,高良姜素+阿帕替尼组对胃癌SGC-7901细胞促凋亡作用明显优于20mg/L阿帕替尼组(P<0.05)。Westernblot结果显示,高良姜素+阿帕替尼组Bcl-2相关X蛋白(Bax)、磷酸化蛋白激酶B( p-Akt)、磷酸化丝裂原活化蛋白激酶(p-p38)蛋白表达明显高于20mg/L阿帕替尼组,B淋巴细胞瘤-2( Bcl-2)蛋白明显低于20mg/L阿帕替尼组(P<0 .05)。结论高良姜素可能通过抑制PI3K/Akt和激活p38-MAPK信号通路增强阿帕替尼抗胃癌SGC-7901细胞的作用。

白介素33激活P38-MAPK通路保护心肌缺血再灌注损伤

目的:探讨白介素33(IL-33)是否通过调控P38MAPK通路影响心肌缺血再灌(I/R)损伤。方法:应用大鼠心肌I/R模型,将32只成年雄性SD大鼠随机分为4组,对照组、模型组、IL-33组、IL-33+SB230580(P38通路抑制剂)组。检测血清中LDH、CK水平及心肌组织中TNF-α、IL-6、总caspase-3、活化caspase-3、Bcl-2、Bax、磷酸化P38(p-P38)表达。结果 :再灌注4 h后,IL-33可明显降低LDH和CK、TNF-α、IL-6、Bax的表达和caspase-3活化,增加Bcl-2、p-P38表达(均P<0.05);SB230580可以一定程度上减弱IL-33的作用。结论 :IL-33可以通过P38-MAPK通路抑制炎症反应和心肌细胞凋亡保护I/R心肌。

大肠癌组织中P38-MAPK通路对CDX2表达影响的研究

目的:探讨大肠癌发生中尾型同源盒转录因子-2(CDX2)的表达特点及其P38-MAPK通路对CDX2表达的影响。方法:收集我院2011年1月—2013年1月大肠癌患者病理组织共138例,采用荧光定量PCR、Western blot、免疫组织化学等技术检测CDX2在正常大肠组织、癌旁组织、大肠癌组织中的表达;应用Western blot检测CDX2的上游分子P38-MAPK表达。结果:CDX2在大肠癌组织、癌旁组织、正常组织中RNA的相对表达量分别为100.0±6.6、60.8±5.7、40.4±5.4,各组织中CDX2相对表达量依次为100.0±9.8、75.3±6.8、35.8±5.4。p38-MAPK蛋白在大肠癌组织、癌旁组织、正常组织的相对表达量分别为100.0±11.2、58.9±8.7、38.1±6.7。结论:大肠上皮组织中p38-MAPK表达减少,导致CDX2表达减少,是大肠癌发生的重要因素之一。

氧化苦参碱对脓毒症大鼠心肌组织p38-MAPK/caspase-3信号通路的影响

目的观察氧化苦参碱(Oxymatrine,OMT)对脓毒症大鼠心肌组织p38-MAPK/caspase-3信号通路的影响。方法采用盲肠结扎穿孔法(cecal ligation and puncture,CLP)复制大鼠脓毒症模型,随机将48只SD大鼠分为假手术组、OMT对照组、模型(CLP)组和CLP+OMT(52、26、13 mg·kg-1)组。采用TUNEL法检测心肌细胞凋亡指数(AI);RT-PCR法测定大鼠心肌组织p38-MAPK、caspase-3 mRNA的表达;Western blot法测定心肌组织磷酸化p38-MAPK、caspase-3的蛋白表达。结果 52、26、13 mg·kg-1的OMT可明显降低心肌细胞凋亡指数(P<0.05);下调p38-MAPK和caspase-3 mRNA的表达(P<0.05),降低心肌组织磷酸化p38 MAPK、caspase-3含量(P<0.05)。结论 OMT可以通过抑制p38-MAPK/caspase-3信号通路活化,减轻脓毒症引起的心肌细胞凋亡。

游离胆固醇经P38-MAPK信号途径激活未折叠蛋白反应诱导巨噬细胞凋亡

目的:探讨p38-MAPK信号通路对未折叠蛋白反应的影响及在游离胆固醇(FC)诱导巨噬细胞凋亡中的作用。方法:收集小鼠腹腔巨噬细胞进行体外培养,使用100μg/mL乙酰低密度脂蛋白及10μg/mL胆固醇乙酰转移酶抑制剂-58035促进FC聚集,以p38特异性抑制剂SB203580进行干预,Annexin-V和PI双染后流式细胞仪检测细胞凋亡,Western-bolt检测磷酸化p38和CHOP表达。结果:普通培养基孵育的巨噬细胞无磷酸化p38和CHOP表达,只有少量细胞凋亡;而在促进FC聚集条件下孵育的巨噬细胞磷酸化p38和CHOP表达明显,8 h后凋亡细胞为(21.8±0.6)%;使用SB203580干预后无磷酸化p38表达,CHOP表达减少,凋亡细胞为(6.9±0.3)%。结论:FC聚集是诱导巨噬细胞凋亡的重要原因,p38通过激活未折叠反应参与这一过程,而SB203580通过抑制p38活性对FC诱导的巨噬细胞凋亡具有保护作用。

脂联素依赖p38-MAPK途径抑制衣霉素诱导的内质网应激致心肌细胞凋亡

目的通过原代培养SD大鼠的乳鼠心肌细胞建立衣霉素心肌细胞内质网应激损伤模型,观察脂联素对心肌细胞内质网应激致细胞凋亡的作用及其机制。方法采用酶消化法原代培养乳鼠心肌细胞,倒置相差显微镜下观察细胞生长,通过α-肌动蛋白免疫荧光法对培养的心肌细胞进行鉴定。选用原代培养3～4天的心肌细胞,随机分为五组:对照组、1 mg/L衣霉素组、1 mg/L衣霉素+100 mg/L脂联素组、1 mg/L衣霉素+3μmol/LSB203580组及1 mg/L衣霉素+3μmol/L SB203580+100 mg/L脂联素组。实验终止后,在倒置相差显微镜下观察心肌细胞形态变化,通过流式细胞术检测心肌细胞凋亡,用qRT-PCR及免疫荧光法检测内质网应激指标GRP78和CHOP的mRNA及蛋白表达。结果与对照组相比,给予衣霉素后,细胞凋亡率显著增加,GRP78和CHOP的mR-NA及蛋白表达增加。脂联素预处理后给予衣霉素,可较大程度地逆转上述指标变化,细胞凋亡率显著下降,GRP78和CHOP的mRNA及蛋白表达减少;而加用p38-MAPK抑制剂SB203580后脂联素的保护作用明显减弱,凋亡率显著增加,GRP78和CHOP的mRNA及蛋白表达增高,但较单纯衣霉素处理组凋亡率低,GRP78和CHOP的mRNA及蛋白表达也减少。结论衣霉素可使GRP78和CHOP表达增强,启动内质网应激,导致心肌细胞凋亡,脂联素可以通过减轻内质网应激逆转衣霉素所致的心肌细胞凋亡作用,对心肌细胞有保护作用,且这种保护作用部分是通过p38-MAPK途径实现的。

Activation and signaling of the p38 MAP kinase pathway

The family members of the mitogen-activated protein (MAP) kinases mediate a wide variety of cellular behaviors in response to extracellular stimuli. One of the four main sub-groups, the p38 group of MAP kinases, serve as a nexus for signal transduction and play a vital role in numerous biological processes. In this review, we highlight the known characteristics and components of the p38 pathway along with the mechanism and consequences of p38 activation. We focus on the role of p38 as a signal transduction mediator and examine the evidence linking p38 to inflammation, cell cycle, cell death, development, cell differentiation, senescence and tumorigenesis in specific cell types. Upstream and downstream components of p38 are described and questions remaining to be answered are posed. Finally, we propose several directions for future research on p38.

Resveratrol Induces Apoptosis and Autophagy in T-cell Acute Lymphoblastic Leukemia Cells by Inhibiting Akt/mTOR and Activating p38-MAPK

Objective To explore the effects of resveratrol-induced apoptosis and autophagy in T-cell acute lymphoblastic leukemia （T-ALL） cells and potential molecular mechanisms. Methods The anti-proliferation effect of resveratrol-induced, apoptosis and autophagy on T-ALL cells were detected by using MTI- test, immunofluorescence, electronic microscope, and flow cytometry, respectively. Western blotting was performed for detecting changes of apoptosis-associated proteins, cell cycle regulatory proteins and state of activation of Akt, mTOR, p70S6K, 4E-BP1, and p38-MAPK. Results Resveratrol inhibited the proliferation and dose and time-dependent manner. It also induced cyclin-dependent kinase （CDK） inhibitors p21 and induced apoptosis and autophagy in T-ALL cells in a cell cycle arrest at G0/G1 phase via up regulating p27 and down regulating cyclin A and cyclin D1. Western blotting revealed that resveratrol significantly decreased the expression of antiapoptotic proteins （Mcl-1 and Bcl-2） and increased the expression of proapoptotic proteins （Bax, Bim, and Bad）, and induced cleaved-caspase-3 in a time-dependent manner. Significant increase in ratio of LC3-11/LC3-1 and Beclin 1 was also detected. Furthermore, resveratrol induced significant dephosphorylation of Akt, mTOR, p70S6K, and 4E-BP1, but enhanced specific phosphorylation of p38-MAPK which could be blocked by SB203580. When autophagy was suppressed by 3-MA, apoptosis in T-ALL cells induced by resveratrol was enhanced. Conclusion Our findings have suggested that resveratrol induces cell cycle arrest, apoptosis, and autophagy in T-ALL cells through inhibiting Akt/mTOR/p7OS6K/4E-BP1 and activating p38-MAPK signaling pathways. Autophagy might play a role as a self-defense mechanism in T-ALL cells treated by resveratrol. Therefore, the reasonable inhibition of autophagy in T-ALL cells may serve as a promising strategy for resveratrol induced apoptosis and can be used as adjuvant chemotherapy for T-ALL.

白杨素通过下调p38-MAPK磷酸化减轻脓毒症大鼠心肌损伤

目的探讨白杨素对脓毒症模型大鼠心肌损伤的作用机制。方法清洁级SD大鼠100只,按随机字母表法分成对照组、模型组、地塞米松组(0.27mg/kg)、白杨素低、高剂量组(20、40mg/kg),每组20只。模型组、地塞米松组、白杨素低、高剂量组建立脓毒症模型;建模成功后,地塞米松组、白杨素低、高剂量组给予相应药物灌胃,对照组及模型组给予等体积生理盐水。连续给药4周后,测定大鼠心功能指标,苏木精-伊红(HE)染色观察大鼠心肌组织病理学变化,实时荧光定量PCR(qRT-PCR)及蛋白免疫印迹(Western blot)测定大鼠心肌p38、磷酸化MAPK(p-MAPK)mRNA和蛋白水平。结果与对照组比较,模型组大鼠左心室舒张末期内径(LVEDD)[(7.86±0.32)mm比(5.69±0.23)mm]、左心室收缩末期内径(LVESD)[(7.16±0.85)mm比(2.99±0.59)mm]、p38 mRNA[(5.41±0.41)比(0.86±0.36)]、p-MAPK mRNA[(5.54±0.39)比(0.83±0.37)]、p38蛋白[(1.32±0.23)比(0.43±0.18)]、p-MAPK蛋白[(1.26±0.19)比(0.40±0.16)]水平升高(P均<0.05),短轴缩短分数(FS)[(27.56±7.59)%比(49.98±7.96)%]、射血分数(EF)[(41.42±6.63)%比(79.43±6.74)%]水平降低(P均<0.05)。与模型组比较,地塞米松组、白杨素低、高剂量组LVEDD[(5.99±0.28)mm、(7.08±0.35)mm、(6.02±0.34)mm比(7.86±0.32)mm]、LVESD[(3.98±0.59)mm、(5.90±0.64)mm、(3.97±0.63)mm比(7.16±0.85)mm]、p38 mRNA[(1.83±0.43)、(3.52±0.46)、(1.82±0.39)比(5.41±0.41)]、p-MAPK mRNA[(2.35±0.31)、(4.34±0.28)、(2.36±0.39)比(5.54±0.39)]、p38蛋白[(0.81±0.20)、(1.04±0.21)、(0.79±0.17)比(1.32±0.23)]、p-MAPK蛋白[(0.75±0.17)、(0.97±0.19)、(0.73±0.18)比(1.26±0.19)]水平降低(P均<0.05),FS[(42.54±5.59)%、(34.48±5.93)%、(41.43±6.85)%比(27.56±7.59)%]、EF[(65.42±11.74)%、(49.62±12.85)%、(64.61±8.85)%比(41.42±6.63)%]水平升高(P均<0.05)。白杨素高剂量组LVEDD、LVESD、FS、EF水平、p38、p-MAPK mRNA和蛋白水平较低剂量组改善更明显(P均<0.05),与地塞米松组比较无明显差异(P>0.05)。结论白杨素能够减轻脓毒症模型大鼠心肌损伤,减轻心肌细胞炎症反应;其机制可能与白杨素显著抑制脓毒症大鼠心肌细胞p38-MAPK磷酸化有关。

Neuroprotective mechanisms of rutin for spinal cord injury through anti-oxidation and anti-inflammation and inhibition of p38 mitogen activated protein kinase pathway

Rutin has anti-inflammatory, antioxidant, anti-viral, anti-tumor and immune regulatory effects. However, the neuroprotective effects of rutin in spinal cord injury are unknown. The p38 mitogen activated protein kinase （p38 MAPK） pathway is the most important member of the MAPK family that controls inflammation. We assumed that the mechanism of rutin in the repair of spinal cord injury is associated with the inhibition of p38 MAPK pathway. Allen’s method was used to establish a rat model of spinal cord injury. The rat model was intraperitoneally injected with rutin （30 mg/kg） for 3 days. After treatment with rutin, Basso, Beattie and Bresnahan locomotor function scores increased. Water content, tumor necrosis factor alpha, interleukin 1 beta, and interleukin 6 levels, p38 MAPK protein expression and caspase-3 and -9 activities in T8–9 spinal cord decreased. Oxidative stress related markers superoxide dismutase and glutathione peroxidase levels increased in peripheral blood. Rutin exerts neuroprotective effect through anti-oxidation, anti-inflammation, anti-apoptosis and inhibition of p38 MAPK pathway.

肾衰康灌肠液在急性肾缺血再灌注损伤中的作用及对p38-MAPK信号通路的影响

目的:研究肾衰康灌肠液在肾缺血再灌注损伤中的作用及其对p38-MAPK信号通路的影响。方法:采用新西兰大白兔灌肠制备肾衰康灌肠液含药血清,体外培养人肾小管上皮细胞株(HK-2细胞),并将其随机分为缺血再灌注组、PBS组、低剂量干预组、中剂量干预组、高剂量干预组,共5组。PBS组常规培养,对缺血再灌注组和低、中、高剂量干预组行急性缺血再灌注损伤造模。缺血再灌注开始后立即给予低剂量干预组(25%高剂量血清+75%空白血清)、中剂量干预组(50%高剂量血清+50%空白血清)、高剂量干预组(100%高剂量血清)含药血清处理。与此同时,缺血再灌注组和PBS组分别给予等量PBS灌肠后所得的兔血清,PBS组不予以细胞造模处理。操作0、4、8、12、24和48h后,分别采用RT-PCR法检测细胞中P38MAPK基因表达情况,采用Annexin V-FITC/PI联合流式细胞术分析细胞凋亡情况。结果:4h时,低剂量组细胞凋亡率为(13.5±0.2)%,中剂量组为(7.8±2.1)%,高剂量组为(13.3±0.1)%;12h时,低剂量组细胞凋亡率为(14.8±0.3%),中剂量组为(10.3±0.6)%,高剂量组为(12.9±0.9%),均显著低于对照组[4h,(47.8±3.5)%;12h,(45.6±2.2)%]和PBS组[4h,(45.7±1.2)%;12h,(41.6±0.7)%]。含药血清处理模型细胞4h后,与PBS组比较,低、中、高剂量组中MAPK基因表达均显著降低(P<0.05),且从8~12h,MAPK基因表达升高,逐渐恢复至正常水平。结论:肾衰康灌肠液可抑制肾缺血再灌注模型中HK-2细胞凋亡,可能是通过抑制p38-MAPK基因表达实现的。