Dioscin reduces lipopolysaccharide-induced inflammatory liver injury via regulating TLR4/MyD88 signal pathway

We previously reported the effects of dioscin against carbon tetrachloride-, acetaminophen- and alcohol-induced acute liver damage. However, its effect on lipopolysaccharide (LPS)-induced inflammatory liver injury remains unknown. In the present work, liver injury in mice and rats was induced by LPS, and dioscin was intragastrically administered for 7 days. In vitro, the AML-12 cells and HepG-2 cells were treated with LPS after dioscin treatment. The results showed that dioscin not only markedly reduced serum ALT, AST levels and relative liver weights, but also restored cell injury caused by LPS. In mechanism study, dioscin significantly attenuated inflammation through down-regulating the levels of toll-like receptor (TLR) 4, myeloid differentiation factor 88 (MyD88), interleukin-1 receptor-associated kinase 1 (IRAK1), tumor necrosis factor receptor-associated factor 6 (TRAF6), phosphorylated inhibitor of nuclear factor κB kinase (p-IKK), phosphorylated inhibitor of nuclear factor κB alpha (p-IκBα), phosphorylated nuclear factor κB p65 (p-NF-κB p65), high-mobility group protein 1 (HMGB-1), interleukin (IL)-1, IL-6 and tumor necrosis factor-α (TNF-α). TLR4 overexpression was also decreased by dioscin, leading to the markedly decreased levels of MyD88, IRAK1, TRAF6, p-IKK, p-IκBα, p-NF-κB p65 and HMGB-1. Suppression of MyD88 by ST2825 eliminated the inhibitory effects of dioscin on the levels of IRAK1, TRAF6, p-IKK, p-IκBα, p-NF-κB p65, HMGB-1, IL-1β, IL-6 and TNF-α. Our results suggested that dioscin exhibited protective effect against LPS-induced liver injury via altering TLR4/MyD88 pathway, which should be developed as one potent candidate for the treatment of acute inflammatory liver injury in the future.     

基于NF-κB/ICAM-1通路探讨右美托咪定对子痫前期大鼠的神经保护作用

目的　基于核因子-κB（NF-κB）/细胞间黏附分子-1（ICAM-1）通路探讨右美托咪定（DEX）对子痫前期大鼠的神经保护作用。方法　SD孕鼠随机分为对照组,模型组,DEX低、中、高剂量组,每组10只。除对照组外,其余各组孕鼠于妊娠第10~20天每日腹腔注射同型半胱氨酸（Hcy,200 mg/kg）及隔日背部皮下注射谷氨酸单钠（MSG,1 g/kg）制备子痫前期模型,在妊娠第10~20天,DEX低、中、高剂量组分别腹腔注射7.5、15.0、30.0 μg/kg 的DEX。神经行为学实验评估大鼠行为学缺损情况;酶联免疫吸附试验（ELISA）法检测脑脊液中S100B、铁蛋白和脑组织炎性因子白细胞介素（IL）-1β、肿瘤坏死因子-α（TNF-α）及IL-6表达水平;TUNEL染色检测脑组织细胞凋亡情况;Western blot检测大鼠脑组织NF-κB/ICAM-1通路相关蛋白表达。结果　与对照组相比,模型组大鼠脑组织细胞凋亡现象严重,行为学缺损评分,脑脊液中S100B与铁蛋白,脑组织IL-1β、TNF-α、IL-6含量,p-NF-κB/NF-κB、ICAM-1蛋白表达水平显著升高（P＜0.05）;与模型组相比,DEX低、中、高剂量组大鼠凋亡细胞比例较低,行为学缺损评分,脑脊液中S100B与铁蛋白,脑组织IL-1β、TNF-α、IL-6含量,p-NF-κB/NF-κB、ICAM-1蛋白表达水平下降（P＜0.05）,且DEX各组之间呈剂量依赖性。结论　DEX对子痫前期大鼠神经细胞有保护作用,其机制可能与抑制NF-κB/ICAM-1信号激活、炎性因子释放及脑神经细胞凋亡有关。     

血必净对脓毒症大鼠心肌保护作用的研究

目的观察血必净注射液对脓毒症大鼠心肌损伤的保护作用。方法将30只SD大鼠随机分为假手术组（Sham组）、脓毒症模型组（CLP组）及血必净治疗组（XBJ组），每组10只。Sham组只开腹关腹，余同CLP组；CLP组采用盲肠结扎穿孔法造模；XBJ组造模后立即经腹腔注射血必净注射液8ml／kg。观察24h后检测血清肌钙蛋白Ⅰ（TnI）、脑钠肽（BNP）、肿瘤坏死因子-α（TNF-α）、自细胞介素（IL）-1B，留取左心室心肌组织观察超微结构的改变。结果①脓毒症状态下TnⅠ、BNP、TNF-α、IL-1β水平显著升高，应用血必净注射液后各项指标明显改善。②血必净组心肌组织超微结构的损害较脓毒症组明显减轻。结论血必净注射液能改善脓毒症大鼠的心肌损伤，其机制可能与下调TNF-α、IL-1β水平有关。     

Protocatechuic aldehyde protects against experimental sepsis in vitro and in vivo

Recent studies have demonstrated that nuclear factor-κB (NF-κB) and high-mobility group box 1 (HMGB1) are associated with the pathophysiology of sepsis. The present study was carried out to investigate the effects of protocatechuic aldehyde (PA) on an experimental model of sepsis induced by caecal ligation and puncture (CLP) in rats and to elucidate the potential mechanism in the cultured murine macrophage cell line, RAW264.7 cells. Treatment of RAW 264.7 cells with PA blocked TNF-α-induced NF-κB phosphorylation and decreased HMGB1 expression. Septic rats received doses of 50 mg of PA alone or plus Imipenem by intravenous bolus injection into the tail vein. The results showed that PA reduced serum levels of HMGB1 and triggering the receptor expressed on myeloid cells, it attenuated myeloperoxidase in the lung, liver and small intestine, while it up-regulated serum level of IL-10. Meanwhile, PA alone or plus Imipenem reduced CLP-induced lethality in septic rats. These data indicate that the anti-septic effect of PA is mediated by decreasing local and systemic levels of a wide spectrum of inflammatory mediators. The protective effects of PA might block the inflammatory cascades through HMGB1 and NF-κB signalling pathway. Our studies enhance the case for the use of PA in sepsis, and PA therefore seems promising in the treatment of sepsis in human beings.     

mTOR-自噬通路对脓毒症小鼠海马小胶质细胞表型的影响

目的 探讨哺乳动物雷帕霉素靶蛋白（mTOR）-自噬通路对脓毒症小鼠海马小胶质细胞表型的影响。方法 54 只雄性 ICR 小鼠,按照随机数字表法分为 3 组（n=18）：假手术组（Sham 组）、脓毒症组（CLP 组）和脓毒症+mTOR抑制剂雷帕霉素组（CLP+Ra组）。Sham组只进行开腹手术操作;CLP组采用盲肠结扎穿孔术制备脓毒症小鼠模型;CLP+Ra 组于造模前 6 h腹腔注射雷帕霉素（1.5 mg/kg）。于造模后 24 h各组取 6只,采用酶联免疫吸附试验（ELISA）法检测海马组织肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β、IL-10和转化生长因子（TGF）-β水平;采用免疫荧光染色法检测海马组织切片离子钙接头蛋白分子（Iba）-1/CD86和 Iba-1/CD206阳性细胞数量;采用蛋白免疫印迹法测定海马组织 mTOR磷酸化（p-mTOR）水平、自噬相关蛋白微管相关蛋白 1轻链 3Ⅱ（LC3Ⅱ）和 Beclin-1表达情况。结果 与 Sham组比较,CLP组 TNF-α、IL-1β、IL-10和 TGF-β水平升高,Iba-1/CD86和 Iba-1/CD206阳性细胞数量增加,p-mTOR水平上调,LC3Ⅱ和 Beclin-1表达下调（均 P＜0.05）。与 CLP组比较,CLP+Ra组 TNF-α和 IL-1β水平降低,IL-10和 TGF-β水平升高,Iba-1/CD86阳性细胞数量减少,Iba-1/CD206阳性细胞数量增加,p-mTOR水平下调,LC3Ⅱ和 Beclin-1表达上调（均 P＜0.05）。结论 mTOR-自噬通路通过调节小胶质细胞表型转化,影响脓毒症小鼠海马炎症反应。     

金骨莲胶囊对炎症模型大鼠的抗炎作用及机制研究

目的:研究金骨莲胶囊对炎症模型大鼠的抗炎作用及机制。方法:将48只大鼠随机分为空白对照组、模型组、金骨莲胶囊低、中、高剂量组(0.66、1.32、2.64 g/kg)和地塞米松组(阳性对照,0.945 mg/kg),每组8只。空白对照组和模型组大鼠灌胃等体积水,其余各组大鼠灌胃相应药物,每天给药2次,连续3天。末次给药后30 min,模型组和给药组大鼠腹腔注射脂多糖(10 mg/kg)以复制炎症模型。腹腔注射6 h后,于大鼠尾静脉取血,采用酶联免疫吸附试验检测大鼠血清中肿瘤坏死因子(TNF-α)、白细胞介素1β(IL-1β)、IL-6、前列腺素E(2 PGE2)的含量;测定大鼠肺组织湿质量/干质量(W/D)比值;采用苏木精-伊红染色法观察大鼠肺组织的病理学形态变化;采用逆转录-聚合酶链式反应法检测大鼠肺组织中TNF-α、IL-6、PGE2、IL-1βmRNA的表达水平;采用Western blot法检测大鼠肺组织中核因子κB(NF-κB)p65蛋白的磷酸化水平和NF-κB抑制蛋白α(IκBα)蛋白的表达水平。结果:与空白对照组比较,模型组大鼠血清中TNF-α、IL-1β、IL-6、PGE2的含量,肺组织W/D比值,肺组织中TNF-α、IL-1β、IL-6、PGE2 mRNA的表达水平和NF-κB p65蛋白磷酸化水平均显著升高,IκBα蛋白表达水平显著降低(P<0.05或P<0.01);大鼠肺组织中大量肺泡萎缩塌陷、壁增厚,可见肺实质化及大量炎症细胞浸润。与模型组比较,金骨莲胶囊各剂量组大鼠血清中TNF-α、IL-1β(低剂量组除外)、IL-6、PGE2的含量,以及肺组织中TNF-α(低剂量组除外)、IL-1β、IL-6(低、高剂量组除外)、PGE2 mRNA的表达水平均显著降低(P<0.05或P<0.01);金骨莲胶囊高剂量组大鼠肺组织W/D比值显著降低(P<0.05或P<0.01);金骨莲胶囊中剂量组大鼠肺组织中NF-κB p65蛋白磷酸化水平均显著降低(P<0.05或P<0.01),IκBα蛋白表达水平显著升高(P<0.05);大鼠肺泡结构较为清晰、壁轻度增厚,并见少量炎症细胞浸润。结论:金骨莲胶囊对炎症模型大鼠具有良好的抗炎作用,其作用机制可能与抑制NF-κB信号通路有关。     

乌司他丁调控脓毒症大鼠TNF-α与IL-6及IL-10水平的研究

目的:探讨乌司他丁对脓毒症大鼠血清TNF-α、L-6、IL-10水平的调控作用。方法:将雄性SD大鼠随机分为正常对照组、假手术组、模型组和乌司他丁组。采用盲肠结扎穿孔术(CLP)制备脓毒症模型,于造模后2.0,8.0,12.0,24.0h处死大鼠,双抗体夹心酶联免疫吸附法(ELISA)测定血清TNF-α、L-6、IL-10水平;留取完整左肺组织行病理切片,评估肺脏损伤情况。结果:模型组各时间点TNF-α、L-6、IL-10水平均高于假手术组(P<0.05);乌司他丁组与模型组相比,除24.0h外TNF-α水平明显降低(P<0.01),IL-6水平各时间点均明显下降(P<0.01),IL-10水平各时间点均明显升高(P<0.01),病理切片显示乌司他丁组肺损伤轻于模型组。结论:乌司他丁能下调脓毒症大鼠TNF-α、L-6水平,对IL-10有上调作用,并能减轻脓毒症肺损伤。     

Anti-inflammatory effect of Yam Glycoprotein on lipopolysaccharide-induced acute lung injury via the NLRP3 and NF-κB/TLR4 signaling pathway

Acute lung injury (ALI) is a common lung disease accompanied by acute and persistent pulmonary inflammatory response syndrome, which leads to alveolar epithelial cells and capillary endothelial cell damage. Yam glycoprotein, separated from traditional Chinese yam, has been shown to have anti-inflammatory and immunomodulatory effects. In this experiment, we mainly studied the therapeutic effect and mechanism of a glycoprotein on the lipopolysaccharide (LPS)-induced ALI mice. An oral glycoprotein method was used to treat the mouse ALI model induced by LPS injection in the peritoneal cavity. Afterward, we measured the wet/dry (W/D) ratio, the activity of myeloperoxidase (MPO), the oxidative index superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-PX) and the production of inflammatory cytokines interleukin-1β (IL-1β), tumour necrosis factor-α (TNF-α), and interleukin-6 (IL-6) to evaluate the effect of yam glycoprotein on lung tissue changes. We examined the protein expression of TLR4, ASC, NF-κBp65, p-NF-κBp65, Caspase-1, IκB, NLRP3, p-IκB, and β-actin by western blot analysis. Immunohistochemical analyses of NLRP3 and p-p65 in lung tissue were carried out to assess the mechanism of glycoprotein action. This result suggests that glycoprotein markedly depressed LPS-induced lung W/D ratio, MPO activity, MDA content SOD and GSH-Px depletion, and the contents of inflammatory cytokines IL-1β, IL-6, and TNF-α. Moreover, glycoprotein blocked TLR4/NF-κBp65 signaling activation and NLRP3inflammasome expression in LPS-induced ALI mice. As this particular study shows, glycoprotein has a safeguarding effects on LPS-induced ALI mice, possibly via activating NLRP3inflammasome and TLR4/NF-κB signaling pathways.     

洛伐他汀对脓毒症大鼠肺损伤的保护作用及对脂联素表达的影响

目的：观察洛伐他汀对脓毒症大鼠肺损伤的保护作用及对脂联素表达的影响。方法健康雄性清洁级Wistar大鼠54只,体质量250～300 g,采用数字表法随机分为三组：假手术组（Sham组）、脓毒症组（CLP组）、洛伐他汀处理组（LOV组）。 Sham组及LOV组术前1周腹腔注射洛伐他汀4 mg／kg,CLP组则注射等体积的溶剂0．5％缩甲基纤维素钠（CMC）。各组术后4 h、12 h和24 h取支气管肺泡灌洗液（BALF）测定TNF-α及IL-6水平;取肺组织观察组织病理学改变及检测髓过氧化物酶（MPO）及丙二醛当量（MDA）含量;另取血标本检测血清脂联素水平。结果 Sham组4 h、12 h和24 h TNF-α含量分别是（1．80±0.13）pg／mL、（2．04±0．15）pg／mL和（1．93±0．19）pg／mL;IL-6含量分别是（20．56±0．23）pg／mL、（18．35±0．15）pg／mL和（21．23±0．20）pg／mL;MPO含量分别是（2．82±0．14）U／g组织、（2．88±0．07）U／g组织和（2.76±0．18）U／g组织;MDA含量分别是（3．32±0．12）nmol／mg、（3．09±0．11）nmol／mg和（3．21±0．08）nmol／mg;脂联素浓度分别是（2．68±0．14）μg／mL、（2．80±0．07）μg／mL和（2．86±0．18）μg／mL。与Sham组相比：（1）CLP组及LOV组肺损伤形态学改变明显,CLP组BALF中TNF-α含量[4 h、12 h和24 h分别是（4.23±0．18） pg／mL、（5．62±0．24）pg／mL和（5．14±0．10）pg／mL,t＝28．41、30．98、36．62]及IL-6含量[4 h、12 h和24 h分别是（39．12±0．17）pg／mL、（47．25±0．21）pg／mL和（44．69±0．27）pg／mL,t＝158．90、273．40、127．28]增高,肺组织匀浆中MPO水平[4 h、12 h和24 h分别是（4．85±0．13） U／g组织、（6．17±0.08） U／g组织和（7．84±0．10）U／g组织,t＝26．39、79．40、60．43]及MDA水平[4 h、12 h和24 h分别是（6．24±0．06）nmol／mg、（7．56±0.15）nmol／mg和（8．43±0．10）nmol／mg,t＝53．31?     

低氧诱导因子-1α对脓毒症肠黏膜屏障的保护作用

目的:探讨低氧诱导因子-1α(HIF-1α)对脓毒症肠黏膜屏障的影响及机制。方法:SD大鼠随机分4组:假手术组、脓毒症组、脓毒症+HIF-1α刺激剂组(脓毒症+DMOG组)、脓毒症+HIF-1α抑制剂组(脓毒症+Bay87-2243组),每组6只。采用盲肠结扎穿孔(cecal ligation and perforation,CLP)建立脓毒症模型。ELISA检测大鼠血浆炎性标记物IL-1β、IL-6、TNF-α,氧化应激标记物丙二醛(MDA)以及抗氧化因子超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的水平。Western blot检测大鼠肠黏膜HIF-1α表达。HE染色检测肠黏膜病理学损伤。结果:与假手术组相比,脓毒症大鼠炎症因子、氧化应激因子以及HIF-1α显著上调(P<0.05);给予腹腔注射DMOG明显降低脓毒症大鼠血浆IL-1β、IL-6、TNF-α、MDA水平(P<0.05);增加血浆SOD、CAT水平(P<0.05);HIF-1α表达上调(P<0.05),肠黏膜病理损伤减轻,Chiu's评分显著降低(P<0.05)。口服灌胃Bay87-2243则得到相反的结果。结论:HIF-1α对脓毒症肠黏膜损伤具有保护作用,其机制可能与缓解脓毒症炎性反应和抑制氧化应激水平有关。     

熵权法结合星点设计-效应面法优化祛湿清肺方提取工艺及体外抗炎活性评价

目的:优化祛湿清肺方提取工艺,并对祛湿清肺方提取液进行体外抗炎活性评价。方法:以绿原酸、虎杖苷、黄芩苷、汉黄芩苷、黄芩素、芦荟大黄素、汉黄芩素、大黄素成分含量及得膏率为指标,加水量及提取时间为考察因素,采用熵权法结合星点设计-效应面法对祛湿清肺方提取工艺进行优化。以脂多糖(LPS)诱导大鼠腹腔巨噬细胞(RAW_264.7)为炎症模型,酶联免疫吸附法测定白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、一氧化氮(NO)的含量,蛋白质印迹法检测磷酸化NF-κB抑制蛋白激酶(phosphorylated inhibitor of NF-κB kinase,p-IKK)、磷酸化NF-κB p65(phosphorylated NF-κB p65,p-NF-κB p65)、NF-κB抑制蛋白α(inhibitor of NF-κBα,IκBα)蛋白表达水平变化,评价祛湿清肺方提取液抗炎活性。结果:星点设计-效应面法优化所得的最佳提取工艺为加水量13倍,提取2次,每次提取时间105 min。祛湿清肺方提取液能降低IL-6、IL-1β、TNF-α、NO的含量,抑制p-IKK、p-NF-κB p65蛋白表达,促进IκBα蛋白表达,具有较好的体外抗炎活性。结论:熵权法结合星点设计-效应面法优选的祛湿清肺方提取工艺稳定可行,所得提取液具有较好的抗炎活性,为祛湿清肺方开发和现代化研究提供参考奠定基础。     

What is the role of renin inhibition during rat septic conditions: preventive effect of aliskiren on sepsis-induced lung injury

Sepsis and sepsis-related acute lung injuries (ALIs) are one of the main causes of death among hospitalized patients. Renin–angiotensin–aldosterone system (RAAS) has been reported to have role in sepsis. However, there is no study on aliskiren, a renin inhibitor, on sepsis-induced ALI. We aimed to investigate the potential protective effects of aliskiren in a model of cecal ligation and puncture (CLP)-induced lung injury. The rats were separated into five groups: sham, CLP, CLP?+?aliskiren 50 mg/kg (per orem (p.o.)), CLP?+?aliskiren 100 mg/kg (p.o.), and sham?+?aliskiren 100 mg/kg (p.o.). CLP model was applied via ligation of cecum and two punctures. After experiment, biochemical, molecular, and pathologic examinations were performed on lung and serum samples of rats. In our study, sepsis decreased superoxide dismutase (SOD) and glutathione (GSH) and increased malondialdehyde (MDA) in lung tissues of rats while aliskiren increased the SOD and GSH and decreased MDA. Also, sepsis caused a significant increase in pro-inflammatory cytokine levels (TNF-α, IL-1β, and IL-6) while aliskiren administration decreased these cytokines. Also, aliskiren administration at high dose protected lungs in pathologic evaluation. As a result of RAAS inhibition, aliskiren caused a decrease in serum angiotensin II level and increase in serum renin level. In light of these observations, we can suggest that the therapeutic administration of aliskiren prevented oxidative stress changes and cytokine changes and also protected lung tissues during CLP-induced sepsis by changing status of RAAS.     

Pterostilbene alleviates polymicrobial sepsis-induced liver injury: Possible role of SIRT1 signaling

Liver injury occurs frequently during sepsis. Pterostilbene (Pte), a natural dimethylated analog of resveratrol from blueberries, exerts anti-inflammatory and anti-apoptotic effects in various diseases. However, the role of Pte in sepsis-induced liver injury and its underlying mechanisms remain unknown. The current study aimed to evaluate the protective effects of Pte on sepsis-induced liver injury and its potential mechanisms. Sepsis was induced using cecal ligation and puncture (CLP) in C57BL/6 mice. Mice were administered Pte (5, 10, 15 mg/kg, i.p.) at 0.5 h, 2 h, and 8 h after CLP induction. The pathological changes of the liver were evaluated using hematoxylin and eosin (H&E) staining. The serum levels of alanine transaminase (ALT) and aspartate aminotransferase (AST) were measured. The levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL-6), myeloperoxidase (MPO), p38 mitogen-activated protein kinase (p38MAPK), Bax, and B-cell lymphoma 2 (Bcl-2) were also evaluated. Pte treatment attenuated the CLP-induced liver injury, as evidenced by the attenuated histopathologic injuries and the decreased serum aminotransferase levels. Pte reduced the serum inflammatory cytokine (TNF-α and IL-6) levels and hepatic mRNA levels of TNF-α and IL-6. Pte also reduced MPO activity and p38MAPK activation in the liver. Additionally, Pte significantly inhibited Bax expression and increased Bcl-2 expression. Moreover, Pte increased the expression of sirtuin-1 (SIRT1) and reduced the expression of acetylated forkhead box O1 (Ac-FoxO1), acetylated Ac-p53, and acetylated nuclear factor-kappa beta (Ac-NF-κB). However, SIRT1 small interfering RNA (siRNA) abolished Pte's effects on the expression levels of those protein. Notably, Pte improved the survival rate in septic mice. In conclusion, Pte alleviates sepsis-induced liver injury by reducing inflammatory response and inhibiting hepatic apoptosis, and the potential mechanism is associated with SIRT1 signaling activation.     

Characterization, epitope identification and mechanisms of the anti-septic capacity of monoclonal antibodies against macrophage migration inhibitory factor

Sepsis is characterized by uncontrolled inflammatory responses. Macrophage migration inhibitory factor (MIF) has been shown to play an important role in the progression of sepsis thus is a potential therapeutic target. The aim of this study is to produce IgG anti-MIF monoclonal antibodies (mAbs) with anti-septic abilities in vivo and to determine mechanisms of their function. We generated 8 IgG anti-MIF mAbs with high specificity and 3 of them showed potent protective abilities in murine lethal peritonitis induced by cecal ligation and puncture (CLP). One anti-MIF mAb, F11, showed 100% protection within 72 h after sepsis induction and 72% mice treated with this mAb survived up to 84 h with reduced lung and kidney pathology. F11 treatment also reduced tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels in septic mice. We further found that all 8 anti-MIF mAbs recognized the same epitope located in the amino acid residue 1-20 region of the N terminus of the MIF protein. Three of the mAbs, F11 in particular, inhibited tautomerase activity in association with their protective effect on CLP mice. Thus, we have produced anti-MIF mAbs that protected mice from CLP-induced sepsis by recognizing the same epitope domains in MIF. These mAbs are promising candidates for further development of therapeutics against inflammatory diseases.     

辛伐他汀对脓毒症小鼠肺损伤的保护作用

目的观察辛伐他丁对脓毒症小鼠急性肺损伤的保护作用,并探讨其可能机制。方法采用盲肠结扎穿孔术制备脓毒症小鼠模型,将72只雄性C57BL/6小鼠随机分成3组:假手术组、脓毒症急性肺损伤组(脓毒症组)、脓毒症+辛伐他汀治疗组(治疗组)。治疗组给予辛伐他汀0.2μg/g,q12h腹腔注射1周;假手术组、脓毒症组给予等量安慰剂腹腔注射1周。分别于造模后6、12、24 h留取肺脏标本。HE染色观察肺组织病理学变化,免疫组化检测肺组织toll样受体4(Toll-like receptor 4,TLR4)蛋白的表达,ELISA测定肺组织匀浆中IL-1β及TNF-α的表达水平。结果与假手术组比较,造模后6、12、24 h,脓毒症组肺组织病理学评分、肺组织TLR4蛋白的表达,以及TNF-α、IL-1β水平明显升高(P<0.05);与脓毒症组相比,治疗组上述指标明显降低(P<0.05)。结论辛伐他汀通过抑制TLR4信号转导通路,减少其下游炎症介质TNF-α、IL-1β的释放,对脓毒症导致的急性肺损伤具有一定保护作用。     

穿心莲内酯对盲肠结扎穿孔诱导的脓毒症大鼠心肺功能和炎症损伤的保护作用

摘要：目的　探究穿心莲内酯对盲肠结扎穿孔（CLP）诱导的脓毒症大鼠心肺功能和炎症损伤的保护作用。方法　75只SD大鼠随机分为假手术组（A组）、脂多糖模型组（B组）、CLP组（C组）、CLP+低剂量穿心莲内酯组（D组）和CLP+高剂量穿心莲内酯组（E组）,每组15只。A组仅进行开腹、分离盲肠及关腹手术;B组采用脂多糖腹腔注射建立脓毒症模型,C、D、E组行CLP手术诱导脓毒症,D、E组建模后分别腹腔注射10 mg/kg和20 mg/kg穿心莲内酯。比较各组大鼠心率（HR）、左心室射血分数（LVEF）;采用酶联免疫吸附试验（ELISA）检测心肌肌钙蛋白I（cTnI）、肌酸激酶同工酶MB（CK-MB）;比较各组大鼠肺损伤评分;采用血气分析仪检测氧分压［p（O2）］,计算动脉血氧分压/吸入气氧浓度［PaO2/FiO2］;取左肺组织计算湿/干质量（W/D）比值;HE染色观察心肺损伤情况;Western blot检测心肺组织中半胱氨酸天冬氨酸蛋白酶3（Caspase）-3、Caspase-9、B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）和P65蛋白的表达;ELISA检测外周血中白细胞介素（IL）-6、肿瘤坏死因子（TNF-α）、IL-1β、中性粒细胞趋化因子1（CINC-1）水平;使用全自动血细胞分析仪检测中性粒细胞（NEU）水平。结果　与A组相比,B、C组大鼠HR、LVEF、p（O2）和PaO2/FiO2均显著降低,cTnl、CK-MB、肺损伤评分、肺W/D比值、心肺组织中Caspase-3和Caspase-9蛋白表达、Bax/Bcl-2比值、CINC-1、NEU、IL-6、IL-1β、TNF-α和p-P65/P65水平均显著升高（P＜0.05）。与C组相比,D、E组以上指标变化均相反（P＜0.05）;HE染色示,D、E组大鼠心肌肌纤维断裂溶解减少,肌细胞核溶解、固缩显著减少,肺间质中性粒细胞浸润减轻、肺泡间隔增厚、细胞水肿显著改善。结论　穿心莲内酯可以缓解CLP诱导的脓毒症大鼠体内炎症反应并改善大鼠心肺功能,同时抑制凋亡蛋白的表达,保护大鼠心肺组织。     

细粒棘球蚴囊液通过Toll样受体2调节细胞因子分泌的研究

目的探讨细粒棘球蚴囊液(EgCF)是否通过刺激巨噬细胞高表达Toll样受体2(TLR2),进而调节细胞因子的表达。方法培养获得腹腔巨噬细胞,用EgCF刺激小鼠腹腔巨噬细胞,用实时荧光聚合酶链反应检测不同时间点TLR2 mRNA的表达水平,用Western Blot法检测刺激不同时间TLR2信号通路中磷酸化的P38丝裂原活化蛋白激酶(p-P38)、磷酸化的丝氨酸/苏氨酸蛋白激酶(p-AKT)、磷酸化的细胞外调节蛋白激酶(p-ERK)和磷酸化的核转录因子κB-P65(p-NF-κB-P65)的表达水平。siRNA沉默TLR2后,用EgCF再次刺激腹腔巨噬细胞,用Western Blot法检测p-P38、p-AKT、p-ERK和p-NF-κB-P65的表达水平,用酶联免疫吸附实验法检测白细胞介素-6(IL-6)、IL-10和肿瘤坏死因子-α(TNF-α)的表达水平。结果巨噬细胞在EgCF刺激后TLR2 mRNA及p-P38、p-AKT、p-ERK、p-NF-κB-P65表达水平均升高。沉默TLR2后,p-P38、p-AKT和p-ERK表达水平均降低,但p-NF-κB-P65无改变。沉默组的IL-6和TNF-α表达水平均较EgCF刺激组升高,IL-10较EgCF刺激组降低。结论EgCF刺激巨噬细胞高表达TLR2,可能通过TLR2介导的信号通路抑制促炎因子TNF-α和IL-6分泌,促进抑炎因子IL-10的分泌。     

骨髓间充质干细胞对脓毒症急性肺损伤大鼠肺内NF-κB影响的研究

目的 建立大鼠脓毒症急性肺损伤模型,探讨骨髓间充质干细胞（BMSCs）在脓毒症大鼠急性肺损伤中的肺保护作用及可能机制,为临床治疗脓毒症急性肺损伤提供新的理论和实验依据。方法 全骨髓培养法制备BMSCs。36只成年Wistar大鼠（150±15）g随机分为3组,Sham组、CLP组和BMSCs组,每组12只,CLP组和BMSCs组采用盲肠结扎穿孔术（CLP）建立脓毒症急性肺损伤模型,盲肠根部丝线结扎,盲肠切口5mm,肠内容物漏出,将其回纳入腹腔,关腹。Sham组大鼠仅翻动盲肠,不结扎穿孔。BMSCs组术后经尾静脉注射BMSCs细胞悬液1ml（1×10^6／ml）。实验结束后,应用ELISA测定血浆IL-10、MIP-2水平,左肺采用Western blot测定肺内NF-κB的表达水平,右肺制作病理切片,HE染色,并在光镜下观察肺组织病理结构改变。结果 CLP组的血清MIP-2水平明显高于Sham组和BMSCs组,BMSCs组的血清MIP-2水平明显高于Sham组,差异有统计学意义（P〈0．05）;3组的血清IL-10水平比较差异无统计学意义（P〉0．05）：3组的NF-κB表达水平比较差异有统计学意义（P〈0．05）。肺组织病理结果 示CLP组和BMSCs组肺内大量炎症细胞浸润、肺间质水肿、肺内出血,BMSCs组症状较CLP组明显减轻。结论 脓毒症急性肺损伤时。血浆MIP-2表达明显升高,肺内出血、水肿、大量炎症细胞浸润。BMSCs通过调控肺内炎症细胞NF-κB入核,使其促炎细胞因子MIP-2表达减少。减少中性粒细胞浸润起肺保护作用,暂不能说明BMSCs对IL-10有影响。     

己酮可可碱弱化大鼠脓毒症急性肺损伤对p38MAPK/IκBα磷酸化的影响

目的探讨己酮可可碱（pentoxifylline,PTX）对腹腔感染致脓毒症急性肺损伤发挥保护作用的上游信号机制。方法采用盲肠结扎穿孔（cecal ligation and puncture,CLP）致脓毒症模型,将大鼠随机分为假手术组、脓毒症组、脓毒症＋西黄著胶组、脓毒症＋生理盐水组、脓毒症＋SB203580组、脓毒症＋PTX组。检测假手术组、脓毒症组各时间点（1,3,6,12,24 h）p38MAPK及IκBα的磷酸化,然后选择1,6,24 h组分别检测应用SB203580或PTX后p38MAPK及IκBα的表达,同时检测血浆TNF-α、IL-6的含量并观察24 h组肺组织病理改变。结果与假手术组比较,脓毒症组在各个时间点p38MAPK及IκBα均有较强的表达,SB203580或PTX预处理后各组的p38MAPK及IκBα的磷酸化明显受到抑制,且与血浆TNF-α、IL-6的含量以及肺的病理切片变化一致。结论 PTX对脓毒症急性肺损伤的保护作用以及抑制促炎因子产生的作用可能是通过抑制p38MAPK的磷酸化从而抑制NF-κB的活化而产生的。