盐酸戊乙奎醚抑制脂多糖致急性肺损伤大鼠肺组织中PMN扣押和NF-κB活化

目的：观察盐酸戊乙奎醚(PHC)对脂多糖(LPS)致急性肺损伤(ALI)大鼠中性粒细胞(PMN)肺内扣押及对肺组织核因子κB（NF-κB）活化的影响。方法： SD大鼠随机分为对照组、LPS模型组（静脉注射5 mg/kg LPS）、LPS+PHC高、中和低(3.0、1.0和0.3 mg/kg)3个剂量组，每组8只，用比色法测定肺组织髓过氧化物酶(MPO)活性，进行支气管肺泡灌洗液(BALF)PMN计数，蛋白免疫印迹法检测肺组织NF-κB的表达。结果： PHC显著降低ALI大鼠肺组织MPO活性、BALF中PMN计数比例（均P＜0.05)；ALI组大鼠肺组织磷酸化NF-κB的表达显著高于正常对照组（P＜0.05)；PHC高、中剂量组能显著抑制大鼠肺组织磷酸化NF-κB表达高于ALI模型组（均P＜0.05)；在造模后不同的时点观察，PHC对磷酸化NF-κB表达的作用有差别，以造模后6 h时最能有效抑制磷酸化NF-κB上调。结论： PHC能抑制LPS诱导ALI大鼠PMN在肺内扣押和肺组织NF-κB活化，PHC抑制LPS诱导PMN肺内扣押可能与抑制NF-κB活化有关，后者有待进一步验证。     

盐酸戊乙奎醚抑制脂多糖性肺损伤大鼠肺泡II型上皮细胞损伤和ERK活化

目的： 观察盐酸戊乙奎醚(PHC)对脂多糖(LPS)性肺损伤(ALI)大鼠肺泡II型上皮细胞(ATII)和肺组织细胞外信号调节激酶（ERK）活化的影响。 方法： SD大鼠随机分为对照组、LPS组（静脉注射5 mg/kg LPS）和LPS+PHC高、中、低(3.0、1.0和0.3 mg/kg)3个剂量组,每组8只,测定肺湿重/干重（W/D）比值,考马斯亮蓝法测BALF蛋白含量,双缩脲法测血浆蛋白含量,并计算肺通透指数（LPI=BALF蛋白/血浆蛋白）,透射电镜观察各组ATII超微结构,并进行PHC影响肺组织ERK表达的量效性分析;另取大鼠在注入生理盐水（NS）后即刻0 h（对照组）和注射LPS后2 h、4 h、6 h和12 h共5个时点,每时点6只,进行PHC影响肺组织ERK表达的时效性分析。蛋白免疫印迹法检测肺组织ERK的表达。 结果： LPS组大鼠电镜下可见ATII板层小体排空明显而致空泡化,微绒毛脱落,微丝模糊、断裂、缩短,线粒体空泡变性,基膜不完整,而PHC显著减少ATII板层小体空泡化,减轻ATII损伤。LPS模型组大鼠肺W/D比值、LPI及肺组织磷酸化ERK表达显著高于对照组（均P＜0.05),PHC高剂量组显著降低LPS诱导的大鼠肺W/D比值、LPI（均P＜0.05),显著抑制LPS诱导的大鼠肺组织磷酸化ERK表达（P＜0.05);PHC在LPS注射后6h时最能有效抑制ERK磷酸化。结论： PHC抑制LPS诱导的ALI大鼠肺组织通透性增加、ATII损伤和肺组织ERK活化,PHC对LPS诱导大鼠肺组织通透性增高和ATII损伤的拮抗作用可能与抑制ERK活化有关。     

异丙酚抑制脂多糖致大鼠脑损伤时p38 MAPK的活化

目的: 观察异丙酚对脂多糖(LPS)致大鼠脑损伤时脑组织p38丝裂原活化蛋白激酶(p38 MAPK)和诱导型一氧化氮合酶(iNOS)活化的影响。 方法: SD大鼠,雌雄不限,随机分为3组(n=24):对照组(C组)、LPS组(LPS组)和LPS+异丙酚组(LPS+P组),LPS组经左颈内动脉注射LPS(1 mg/kg)建立LPS性脑损伤模型,LPS+P组在给予LPS后立即通过腹腔注射给予异丙酚(100 mg/kg),C组给予等容量生理盐水。分别于注射异丙酚后6、24 、48和72 h处死6只大鼠,取大脑皮质组织,测定脑组织含水量、磷酸化p38 MAPK和iNOS表达水平,光镜下观察脑组织形态及病理变化。 结果: 与C组比较,LPS组各时点脑组织含水量升高,脑组织磷酸化p38 MAPK和iNOS表达水平均于6 h开始增加,24 h达高峰,48 h及72 h各指标仍高于C组(P<0.05);与LPS组相比,LPS+P组脑组织含水量、磷酸化p38 MAPK和iNOS的表达水平降低(P<0.05)。脑组织含水量与磷酸化p38 MAPK、iNOS水平呈正相关(r=0.603,r=0.727, P<0.05)。LPS+P组脑组织病理学损伤轻于LPS组。 结论: 异丙酚可减轻LPS所致的大鼠脑损伤,其机制可能与异丙酚抑制脑组织p38 MAPK活化,下调iNOS的表达,进而减轻炎性反应有关。     

右美托咪啶通过抑制p38和JNK活化减少异氟醚诱导的新生大鼠海马神经元凋亡

 目的：探讨右美托咪啶（dexmedetomidine, Dex）对异氟醚（isoflurane, Iso）诱导的新生大鼠海马神经元凋亡的影响及其与p38丝裂原活化蛋白激酶（p38 mitogen-activated protein kinase,p38）和c-Jun 氨基末端激酶（c-Jun N-terminal kinase, JNK）蛋白活化的关系。方法：48只出生后7 d的SD大鼠随机分为对照组（Con组）、Dex组、Iso组和Iso+Dex组。前2组吸入空气,后2组吸入0.75% Iso 6 h。Dex组和Iso+Dex组在麻醉前20 min和麻醉开始后2 h、4 h经腹腔注射25 μg·kg-1 Dex;Con组和Iso组则在相同的时点注射150 μL生理盐水。麻醉结束后使用TUNEL法检测海马神经元凋亡; Western blotting检测海马组织激活型caspase-3、p38、磷酸化p38（p-p38）、JNK和磷酸化JNK（p-JNK）蛋白表达的变化。结果：(1) Iso组海马CA1区TUNEL阳性细胞数较Con组增加447.57%（P<0.01）,Dex抑制Iso诱导的TUNEL阳性细胞增加达75.18%（P<0.01）。(2) Iso组海马组织激活型caspase-3的表达比Con组增加126.29% （P<0.01）;Dex显著抑制了Iso诱导的caspase-3活化（P<0.01）。(3) Iso组海马p-p38/p38和p-JNK/JNK比值均较Con组明显增加（P<0.01）;Dex显著抑制了Iso诱导的p-p38 和p-JNK表达增加（P<0.01）。结论：Dex能通过减少海马神经元凋亡来减轻Iso对新生大鼠的脑毒性。抑制p38和JNK的磷酸化可能是Dex发挥保护作用的机制之一。     

糖皮质激素通过抑制p38MAPK信号通路缓解大鼠内毒素性急性肺损伤

目的： 研究糖皮质激素对内毒素导致的急性肺损伤的影响及作用机制。方法: 成年雄性SD大鼠被随机分为6组：对照组（control 组,n=6）;LPS组（n=24）;地塞米松＋LPS组（Dex＋LPS组,n=24）;糖皮质激素受体拮抗剂RU486组（RU486组,n=6）;RU486+LPS组（n=24）以及RU486＋Dex＋LPS组（n=24）。除对照组和RU486组外,其余4组在注射LPS后1、3、6、12 h又被分为4个亚组。分别检测各组大鼠支气管肺泡灌洗液（BALF）中TNF-α和IL-6的浓度,肺组织的病理变化,以及肺组织中p38MAPK的活化状态和MKP-1的表达情况。另外又分别比较了LPS组与RU486＋LPS组、Dex＋LPS组与RU486＋Dex＋LPS组大鼠48 h的死亡率。结果: 注射LPS后,BALF中TNF-α 和IL-6的浓度明显升高（P<0.05）,HE染色显示肺组织内广泛炎症反应。而这些现象在应用RU486后变得更加严重,并且RU486＋LPS组的死亡率也明显高于LPS组(P<0.05)。地塞米松能明显缓解LPS导致的肺损伤,糖皮质激素受体(GR)参与此作用。另外,在注射LPS后,肺组织中磷酸化的p38MAPK的表达明显升高,而MKP-1的表达则明显受到抑制。地塞米松能显著降低p38MAPK的磷酸化,这一作用也是GR依赖的。结论: 糖皮质激素活化GR诱导肺组织中MKP-1的表达,进而抑制p38MAPK的活化,从而发挥其抗炎作用,缓解LPS诱导的肺损伤。     

一氧化氮激活应激活化蛋白激酶/c-Jun氨基末端激酶及p38MAP激酶诱导脑缺血再灌注后神经元凋亡

目的研究脑缺血区周边组织一氧化氮合酶（NOS）的表达与应激活化蛋白激酶／c—Jun氨基末端激酶（SAPK／JNK）及p38MAP激酶（p38MAPK）激活的关系;探讨一氧化氮（NO）诱导脑缺血再灌注后神经元凋亡的可能机制。方法采用TUNEL染色法观察脑缺血再灌注不同时段模型鼠缺血区周边组织凋亡的阳性神经元数量;免疫组织化学、蛋白免疫印迹方法检测活化型Caspase-3、神经元型一氧化氮合酶（nNOS）、诱导型一氧化氮合酶（iNOS）和SAPK／JNK,p38MAPK及其磷酸化组分的表达。结果再灌注后1h、2h缺血区周边组织nNOS表达明显增强;自1h起iNOS开始表达,12h达到高峰。1hp-SAPK／JNK表达较强,以后逐渐减弱;p38MAPK各时段表达均明显增强,以6h为著,p-p38MAPK表达高峰亦在6h。6h活化型Caspase．3开始表达,12h达到高峰;12h开始出现TUNEL阳性神经元,24h达到高峰。结论缺血区周边组织NOS表达的增强可能通过激活SAPK／JNK及p38MAPK诱导脑缺血再灌注后神经元凋亡。     

正正常肠淋巴液对内毒素休克小鼠多器官损伤的作用*

 目的： 观察正常肠淋巴液对内毒素休克小鼠肺、心、肝等器官损伤以及MAPK信号通路主要信号分子p38 MAPK、ERK1/2和JNK磷酸化水平的影响。方法: 引流18只BALB/c雄性小鼠正常肠淋巴液,去除细胞成分后用于内毒素休克的干预。18只小鼠均分为假休克组、内毒素休克组与肠淋巴液干预组（n=6）;腹腔注射脂多糖（LPS,35 mg/kg）,复制小鼠内毒素休克模型;在LPS注射60 min后,淋巴液干预组小鼠经股动脉注射正常淋巴液（全血量的1/15）;整个实验过程监测平均动脉血压（MAP）;在腹腔注射LPS后6 h或相应时点,心尖穿刺取血,检测反映心肌和肝细胞损伤的生化指标;同时,留取固定位置的肺、心肌和肝组织,部分观察组织形态,部分检测p38 MAPK、ERK1/2和JNK的磷酸化水平。结果: 内毒素休克组与肠淋巴液干预组小鼠在腹腔注射LPS后90 min多个时点的MAP显著低于假休克组,肠淋巴液干预组小鼠MAP在腹腔注射LPS后80 min、90 min、190 min、210 min、240 min、250 min、340 min、350 min和360 min显著高于内毒素休克组。假休克组小鼠肺、肝和心肌组织结构基本正常,内毒素休克组小鼠出现了一定的结构损伤,肠淋巴液干预组小鼠组织损伤较轻;内毒素休克组小鼠血浆AST、ALT和CK-MB活性高于假休克组,肠淋巴液干预组小鼠血浆CK-MB活性亦高于假休克组,AST和LDH-1活性低于内毒素休克组。注射LPS后6 h,小鼠肺组织p38 MAPK、ERK 1/2和JNK的磷酸化水平显著升高,心肌和肝组织的这些指标未见显著变化;输入正常肠淋巴液降低了内毒素休克小鼠肺组织p38 MAPK以及心肌组织p38 MAPK、ERK 1/2和JNK磷酸化水平。结论: 输入正常肠淋巴液可减轻内毒素休克小鼠的器官损伤,降低肺组织p38 MAPK磷酸化水平。     

胡椒碱抑制LPS活化的人结肠腺癌细胞SW480表达IL-8的作用机制研究

目的研究胡椒碱(piperine,PIP)对脂多糖(LPS)活化的人结肠腺癌细胞SW480表达炎症因子的调节效应及可能的作用机制。方法采用WST-1法检测PIP对SW480细胞增殖的作用,利用流式微球捕获蛋白定量技术(cytometric beads array,CBA)检测炎症因子的蛋白表达水平,以实时定量PCR(qRT-PCR)检测炎症因子mRNA的表达水平,免疫印迹法检测p38MAPK信号通路和JNK信号通路的活化水平。结果 PIP处理可剂量依赖性地抑制SW480细胞的增殖,但PIP对细胞的毒性较小;CBA检测结果显示PIP以剂量依赖性方式抑制LPS诱导的SW480细胞中IL-8的分泌;实时定量RT-PCR检测显示,LPS+PIP处理组与LPS刺激组相比,IL-8的mRNA表达水平明显下降;免疫印迹分析表明,PIP可抑制LPS诱导的p38和JNK MAPK信号通路的活化水平。结论 PIP能够抑制LPS激活的人结肠腺癌细胞SW480中IL-8的分泌从而发挥抗炎作用,其机制可能与抑制p38和JNK MAPK信号通路有关。     

活性氧依赖性p38 MAPK活化与心肌细胞内TNF-α表达的关系

目的:探讨丝裂原活化蛋白激酶(MAPKs)亚型p38在脂多糖(LPS)诱导的心肌细胞肿瘤坏死因子-α(TNF-α)表达过程中的作用及其与活性氧(ROS)的关系.方法:采用消化及差速贴壁法培养新生SD大鼠心肌细胞,应用ELISA检测细胞培养上清TNF-α蛋白含量;采用Western blot测定心肌细胞内p38 MAPK的磷酸化水平来反映其活性;细胞内ROS水平采用ROS敏感的荧光探针二氯荧光素二乙酸(DCF-DA)测定.结果:LPS呈时间依赖性升高心肌细胞TNF-α蛋白表达水平和p38活性,与空白对照比较,LPS作用1 h后心肌细胞磷酸化p38水平即显著升高(P<0.01),3 h后培养上清TNF吨蛋白含量明显增加(P<0.05).给予p38活性抑制剂预处理后,培养上清TNF-α蛋白含量明显下降,与LPS组比较有非常显著性差异(P<0.01).LPS作用1 h后,心肌细胞DCF荧光强度也显著升高(P<0.01).与p38活性变化一致,而且给予抗氧化剂氮乙酰半胱氨酸和二亚苯基碘鎓预处理后,心肌细胞磷酸化p38水平明显降低,与LPS组比较差异明显(P<0.05),与空白对照比较无统计学差异.结论:p38MAPK活化是LPS诱导下心肌细胞TNF-α表达的莺要分子机制之一,而ROS水平的升高是心肌细胞p38 MAPK活化的重要前提.     

封闭枯否细胞对LPS诱导激活丝裂原活化蛋白激酶的影响

目的： 利用小鼠LPS血症模型，探讨封闭枯否细胞（KCs）对LPS诱导MAPK信号转导通路的影响。方法： 雄性昆明种小鼠在注射LPS（5 mg/kg）前48 h及24 h，分别静脉注射GdCl₃（10 mg/kg）或等量的生理盐水，于LPS或生理盐水注射后30 min，分别取出肝脏或分离KCs；体外培养小鼠KCs经GdCl₃（100 μmol/L）预处理1 h，加入含LPS（100 μg/L）的DMEM培养基继续孵育30 min，分别检测肝脏及体内外KCs ERK1/2、p38MAPK蛋白表达和磷酸化水平，及GdCl₃对KCs吞噬、分泌功能的影响。结果： GdCl₃可抑制LPS 诱导KCs活化和TNF-α分泌，但不能抑制LPS诱导KCs或肝脏ERK1/2、p38MAPK磷酸化，也不能影响ERK1/2、p38MAPK蛋白表达，KCs 经GdCl₃单独短时间处理（1 h），可使其少量分泌TNF-α。结论： 封闭KCs并不能通过调节细胞内ERK1/2、p38MAPK信号转导途径，抑制LPS诱导的KCs活化及TNF-α释放，而可能是通过其它信号转导途径（JNK、NF-кB、GPCR等）间的相互作用减轻肝损伤。     

Suppression of MAPK and NF-κB Pathways by Limonene Contributes to Attenuation of Lipopolysaccharide-Induced Inflammatory Responses in Acute Lung Injury

The present study aimed to investigate the protective role of limonene in lipopolysaccharide (LPS)-induced acute lung injury (ALI). ALI was induced in mice by intratracheal instillation of LPS (0.5 mg/kg), and limonene (25, 50, and 75 mg/kg) was injected intraperitoneally 1 h prior to LPS administration. After 12 h, bronchoalveolar lavage fluid (BALF) and lung tissue were collected. Limonene pretreatment at doses of 25, 50, and 75 mg/kg decreased LPS-induced evident lung histopathological changes, lung wet-to-dry weight ratio, and lung myeloperoxidase activity. In addition, pretreatment with limonene inhibited inflammatory cells and proinflammatory cytokines including tumor necrosis factor-α, interleukin-1β, and interleukin-6 in BALF. Furthermore, we demonstrated that limonene blocked the phosphorylation of IκBα, nuclear factor-κB (NF-κB) p65, p38 mitogen-activated protein kinase (MAPK), c-Jun NH2-terminal kinase, and extracellular signal-regulated kinase in LPS-induced ALI. The results presented here suggest that the protective mechanism of limonene may be attributed partly to decreased production of proinflammatory cytokines through the inhibition of NF-κB and MAPK activation.     

p38 MAPK-HSP27信号通路在内毒素致大鼠肺损伤中的作用

 目的：观察p38丝裂原激活蛋白激酶（p38 MAPK)-热休克蛋白27（HSP27）信号通路在急性肺损伤病理过程中的变化规律。方法：健康雄性Wistar大鼠（300~320 g）随机分成正常对照组（A组）、急性肺损伤组（B组）及急性肺损伤+SB203580组（C组）。通过腹腔注射内毒素建立急性肺损伤大鼠模型,分别于实验开始后的0、2、4、6、8 h处死各组大鼠。检测支气管肺泡灌洗液（BALF）白细胞介素6（IL-6）、肿瘤坏死因子α（TNF-α）及BALF中蛋白含量。苏木素-伊红（HE）染色检查肺组织病理变化及免疫荧光方法检测内皮细胞内F-actin和G-actin,计算肺湿干重比值（W/D）。检测肺组织中磷酸化p38 MAPK（p-p38 MAPK）及磷酸化HSP27（p-HSP27）的含量。 结果：B组在实验后2 h BALF中蛋白水平和肺W/D开始明显增加,给予内毒素后8 h肺泡上皮肿胀,肺泡壁增宽,肺泡间质和肺泡腔水肿明显,肺泡内炎症细胞、红细胞和蛋白渗出明显增多,表现出急性肺损伤的病理改变。在给予了p38 MAPK抑制剂SB203580后的C组BALF中蛋白水平及肺W/D分别比B组明显减少,肺泡内炎症细胞、红细胞和蛋白渗出、间质与肺泡水肿均较B组减轻。B组均在实验后2 h血清及BALF中TNF-α和IL-6的浓度开始增加,p-p38 MAPK及p-HSP27的肺内表达开始增加,与A组相比有显著差异。B组实验后8 h的F-actin的表达明显比A组实验后0 h及8 h的增加,给予p38 MAPK抑制剂SB203580的C组肺p-HSP27 和F-actin的表达分别比B组明显减少。结论：内毒素可以通过激活p38 MAPK-HSP27信号通路引起急性肺损伤;阻断该信号通路可以减轻肺损伤。     

Different involvement of the MAPK family in inflammatory regulation in human pulmonary microvascular endothelial cells stimulated with LPS and IFN-γ

Pulmonary endothelial injury is central in the pathogenesis of acute lung injury (ALI). The MAPK signaling cascades are generally thought to be involved in the molecular mechanism underlying the ALI development, but their roles in pulmonary endothelial injury is poorly understood. We thus examined the involvement of the MAPK family member in inflammatory responses of human pulmonary microvascular endothelial cells (HPMVECs) stimulated with LPS and IFN-γ. HPMVECs were found to exhibit the upregulation of expression of Toll-like receptor 4 by IFN-γ, resulting in potentiation of inflammatory cytokine release by LPS stimulation. All MAPKs, ERK1/2, JNK, and p38, were activated by simultaneous stimulation with LPS/IFN-γ. JNK activation in cells stimulated with LPS/IFN-γ was significantly potentiated by the two different p38 inhibitors, SB203580 and RWJ67657, suggesting the negative regulation of JNK activation by p38 in HPMVECs. The mRNA and protein expression levels of ICAM-1 were eliminated by the JNK inhibitor, suggesting that ICAM-1 expression is positively regulated by JNK. The p38 inhibitor significantly enhanced ICAM-1 expression. ERK1/2 activation was not responsible for the LPS/IFN-γ-induced ICAM-1 upregulation in HPMVECs. THP-1 monocyte adhesion to HPMVECs under LPS/IFN-γ stimulation was inhibited by the JNK inhibitor and enhanced by the p38 inhibitor. We conclude that, in HPMVECs stimulated with LPS/IFN-γ, JNK mediates ICAM-1 expression that can facilitate leukocyte adherence and transmigration, while p38 MAPK negatively regulates the upregulation of ICAM-1 through inhibition of JNK activation.     

Urinary trypsin inhibitor attenuates lipopolysaccharide-induced acute lung injury by blocking the activation of p38 mitogen-activated protein kinase

Objective

To investigate the protective effect of urinary trypsin inhibitor (UTI) in a rat model of lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the underlying molecular mechanism.

Methods

Rats were randomly assigned into three groups: control group, LPS treatment group and LPS/UTI treatment group. The serum concentrations of tumor necrosis factor (TNF)-?? and interleukin (IL)-10 were measured by ELISA. The expression of p38 mitogen-activated protein kinase (MAPK) in lung tissues was determined by Western blot analysis.

Results

Administration of UTI reduced the lung wet/dry weight ratio and ameliorated the tissue damage. In the LPS/UTI treatment group, levels of TNF-?? were significantly lower than those in the LPS treatment group, while the levels of IL-10 were significantly higher than those in the LPS treatment group. Western blot analysis revealed that UTI inhibited the phosphorylation of p38 MAPK in lung tissues.

Conclusions

UTI attenuates LPS-induced ALI, probably by adjusting the balance between proinflammatory and anti-inflammatory cytokines. The mechanism responsible for the decreased TNF-?? expression may be related to the inhibitory effect of UTI on p38 MAPK activation.     

p38 MAPK 和STAT3参与CCK-8抑制LPS诱导的大鼠促炎症细胞因子生成

 目的：观察八肽胆囊收缩素（CCK-8）是否改善脂多糖（LPS）引起的大鼠细胞因子的变化,并探讨p38丝裂原活化蛋白激酶(p38 MAPK)和信号转导子及转录激活子3(STAT3)的信号转导作用,以及CCK受体（CCK-R）的作用。方法：4组大鼠尾静脉分别注入生理盐水（对照）、LPS (8 mg/kg)、CCK-8(40 μg/kg)和CCK-8(40 μg/kg)+LPS (8 mg/kg), 酶联免疫吸附法（ELISA）检测血清、肺脏及脾脏中肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）和IL-6的变化,Western blotting和免疫荧光双标激光共聚焦显微镜检测肺脏和脾脏磷酸化p38 MAPK和磷酸化STAT3的表达,RT-PCR检测脾脏CCK-R亚型的mRNA表达。结果：CCK-8可显著抑制LPS诱导的TNF-α、IL-1β和IL-6的增加。CCK-8可增加LPS诱导的大鼠肺脏和脾脏磷酸化p38 MAPK和磷酸化STAT3的表达。LPS有诱导CCK-AR及CCK-BR mRNA表达量增加的作用。结论：CCK-8对LPS刺激的大鼠促炎症细胞因子过量产生有抑制作用,p38 MAPK和STAT3可能参与了其信号转导机制。LPS刺激时,CCK-R受体发生正向调节,CCK-8有可能用于治疗全身性感染及其它的炎症性疾病。     

p38 MAPK Signal Pathway Involved in Anti-Inflammatory Effect of Chaihu-Shugan-San and Shen-Ling-Bai-Zhu-San on Hepatocyte in Non-Alcoholic Steatohepatitis Rats

Background

Traditional Chinese Medicine (TCM), has over thousands-of-years history of use. Chaihu-Shugan-San (CSS), and Shen-ling-bai-zhu-San (SLBZS), are famous traditional Chinese herbal medicine formulas, which have been used in China, for the treatment of many chronic diseases.

Materials and Methods

This study investigated the anti-inflammatory effects of CSS and SLBZS on signaling molecules involved in p38 mitogen-activated protein kinase (p38 MAPK), pathway on hepatocytes of non-alcoholic steatohepatitis (NASH), rats induced by high fat diet. SD male rats were randomly divided into 8 groups: negative control group, model control group, high (9.6g/kg/day)/low (3.2g/kg/day)-dose CSS group, high (30g/kg/day)/low (10g/kg/day)-dose SLBZS group, high (39.6g/kg/day)/low (13.2g/kg/day)-dose integrated group. The rats of NASH model were induced by feeding a high-fat diet. After 16, wks, Hepatocytes were isolated from 6, rats in each group by collagenase perfusion. The liver histopathological changes and serum inflammatory cytokines TNF-α, IL-6 were determined. The proteins of TLR4, phosphor-p38 MAPK and p38 MAPK involved in p38 MAPK signal pathway were assayed.

Results

The statistical data indicated the NASH model rats reproduced typical histopathological features of NASH in human. CSS and SLBZS ameliorated lipid metabolic disturbance, attenuated NASH progression, decreased the levels of TNF-α and IL-6 in serum, as well as inhibited TLR4 protein expression, p38 MAPK phosphorylation, and activation of p38 MAPK. In conclusion, CSS and SLBZS might work as a significant anti-inflammatory effect on hepatocyte of NASH by inhibiting the activation of TLR4, p-p38 MAPK and p38 MAPK involved in p38 MAPK signal pathway.

Conclusion

To some extent, CSS and SLBZS may be a potential alternative and complementary medicine to protect against liver injury, alleviate the inflammation reaction, moderate NASH progression.     

乌司他丁对急性肺损伤大鼠TNT-α和IL-10 mRNA及p38 MAPK表达的影响响

 目的 探讨乌司他丁(UTI)对脂多糖（LPS）诱导急性肺损伤（ALI）大鼠的保护作用及分子生物学机制。方法 Wistar大鼠随机分为对照组、模型组(LPS 5mg/kg,iv)和干预组(UTI 50000U/kg,iv),用Real time RT-PCR法检测肺组织TNF-α和IL-10 mRNA表达,用免疫组化染色和Western blot技术检测肺组织中p38 MAPK的表达。结果 模型组0.5、1和3hTNF-αmRNA的表达分别是对照组的78.55±18.99,128.74±34.79和12.29±1.32倍,UTI预后分别降至20.95±1.45（p<0.01）,58.15±11.01（p<0.01）和2.85±0.57（p<0.05）倍。模型组0.5、1和3hIL-10mRNA的表达分别是对照组的20.89±4.60,38.20±8.26和53.26±8.01倍,乌司他丁干预后分别升至66.77±11.18（p<0.05）,97.69±27.00（p<0.01）和128.62±42.30（p<0.01）倍。模型组p38 MAPK的表达在各个时点均明显升高,UTI干预后p38 MAPK的表达减弱(p<0.05)。结论 UTI通过调节细胞因子基因表达发挥其肺保护作用。p38 MAPK信号通路在UTI下调TNF-α mRNA的表达中发挥了作用。