香椿子正丁醇提取物通过激活Nrf2改善高糖引起的肾小球内皮细胞氧化应激

目的探讨香椿子正丁醇提取物(n-butyl alcohol extract of toona sinensis,NBAE)对高糖引起的肾小球内皮细胞氧化应激的影响及机制。方法分别以高糖(high glucose,HG)、HG+NBAE刺激人肾小球内皮细胞,采用DCFDA法检测细胞内活性氧(reactive oxygenspecies,ROS)生成情况,Nitrate/Nitrite Colorimetric法检测细胞内NO的含量,Western blot检测Nrf2、NQO1及HO-1的蛋白表达,免疫荧光方法检测Nrf2、P47phox在细胞内的定位及表达。结果高糖刺激肾小球内皮细胞后出现氧化应激损伤,ROS升高,NO减少,p47phox表达量增高,Nrf2及下游蛋白NQO1、HO-1表达减少;NBAE可明显提高Nrf2的表达,并增加下游蛋白NQO1和HO-1的表达,从而抑制高糖导致的ROS升高,抑制p47phox的表达,并稳定NO的含量。结论NBAE通过激活Nrf2及其下游靶蛋白来改善高糖对肾小球内皮细胞造成的氧化应激损伤。     

胆红素对急性肺损伤大鼠肺血管内皮细胞核因子κB,细胞间粘附分子1表达的影响

目的探讨胆红素对急性肺损伤(ALI)的对抗作用及其对肺血管内皮细胞核因子-κB和细胞间粘附分子1表达的影响.方法用雄性Wistar大鼠(200-250g)30只,随机分为正常对照组、ALI动物模型组(用内毒素制造模型)、胆红素干预组.采用原位杂交技术半定量法和免疫组织化学染色测定肺血管内皮细胞中细胞间粘附分子-1(ICAM-1)mRNA和核因子κB (NF-κB)蛋白的表达.结果 (1)ALI模型组肺血管内皮细胞ICAM-1mRNA表达和NF-κB核染色阳性细胞百分比与正常对照组比较显著升高,(P<0.001);(2)胆红素干预组ICAM-1mRNA表达和NF-κB染色阳性细胞百分比与模型组相比明显减低,(P<0.001、P<0.01),但与正常对照组比较仍较高(P<0.05).结论 ICAM-1和NF-κB在ALI显著增加,胆红素可以抑制ALI动物NF-κB和ICAM-1mRNA的表达,可能是其对抗急性肺损伤的作用机制之一.     

MiR-146a通过炎症参与小鼠糖尿病心肌病的机制研究

目的:探讨microRNA-146a (miR-146a)对小鼠糖尿病心肌病炎症的影响。方法:20只雄性C57BL/6小鼠随机分为对照组和糖尿病心肌模型组,模型组利用链脲佐菌素(STZ,50 mg/kg)腹腔注射建立糖尿病心肌病模型,对照组给予等体积枸橼酸钠缓冲液,建模成功后提取心脏组织RNA,利用qRT-PCR检测心肌组织中miR-146a和炎症因子白介素1β(IL-1β),白介素6(IL-6),单核趋化因子1(MCP-1)以及NF-κB/p65的mRNA表达;依次分离原代心肌细胞,成纤维细胞和内皮细胞,给予高糖处理后,检测细胞中miR-146a,IL-1β,IL-6,MCP-1以及NF-κB/p65的mRNA表达;内皮细胞转染miR-146a mimic或miR-146a inhibitor,观察炎症因子的表达情况。结果:糖尿病心肌病心肌组织中miR-146a水平较对照组降低(P0.05),炎症因子IL-1β,IL-6,MCP-1以及NF-κB/p65的mRNA表达高于对照组(P0.05);与对照组相比,高糖分别处理原代心肌细胞和成纤维细胞后miR-146a表达未改变(P0.05),而高糖降低内皮细胞中miR-146a的表达(P0.05);高糖增加内皮细胞炎症因子IL-1β,IL-6,MCP-1及NF-κB/p65的mRNA水平(P0.05);内皮细胞转染miR-146a mimic后可阻止由于高糖所导致的内皮细胞中IL-1β,IL-6,MCP-1及NF-κB/p65的增加(P0.05),而转染miR-146a inhibitor后,可引起正常对照组细胞中炎症因子IL-1β,IL-6,MCP-1以及NF-κB/p65表达的增加(P0.05)。结论:糖尿病心肌病中miR-146a的降低以及炎症的增加,与内皮细胞受损相关,且可能通过miR-146a影响炎症因子的表达。     

脂多糖对新生小鼠肺血管内皮细胞的影响及机制探讨

该文旨在探讨脂多糖(lipopolysaccharide, LPS)对新生小鼠肺血管内皮细胞的影响及机制。将分离培养的肺血管内皮细胞随机分为空白对照组和LPS组;用10μg/mL的LPS处理细胞后分别于0 h、6 h、12 h、24 h、48 h时间点收集细胞标本进行检测;采用划痕实验观察LPS对肺血管内皮细胞迁移的影响;用荧光定量PCR(Real-time PCR, RT-PCR)检测白介素-1β(IL-1β)、巨噬细胞炎性蛋白-1α(MIP-1α)、单核细胞趋化蛋白-1(MCP-1)、肿瘤坏死因子-α(TNF-α) mRNA水平变化; Western blot检测血管内皮生长因子(VEGF)、血管内皮生长因子受体2(VEGFR2)及核因子κB(nuclear factor kappa-B, NF-κB)相关蛋白P65水平的变化。结果显示,体外分离培养的肺血管内皮细胞成鹅卵石样排列, VIII因子相关抗原和CD31表面抗原荧光染色阳性。划痕实验中, LPS组细胞在12 h的迁移高于对照组(P0.001);荧光定量PCR检测到LPS组分泌的炎症因子IL-1β、TNF-α和趋化因子MIP-1α、MCP-1的mRNA表达明显高于对照组(P0.001); Western blot显示, LPS组与对照组相比, VEGF蛋白表达在24 h、48 h处降低(P0.05), VEGFR2蛋白表达在各时间段都明显降低(P0.001),同时, NF-κB相关蛋白P65活性显著升高(P0.05)。研究表明,脂多糖诱发的炎症反应影响肺血管内皮细胞的发育,其机制可能与NF-κB通路激活,诱导炎症因子、趋化因子表达升高和VEGF/VEGFR2表达下降有关。     

七氟烷对培养的小鼠小胶质细胞炎症因子表达的影响（英文）

目的:探讨七氟烷对培养的小鼠小胶质细胞中炎症因子表达的影响。方法:取新生(2~3天)C57BL/6小鼠,分离小胶质细胞,将其随机分为4组(n=10):对照组(Control);七氟烷组(Sevoflurane);NF-κB抑制剂组(PDTC);NF-κB抑制剂+七氟烷组(PDTC+Sevoflurane)。用Drager麻醉机向Sevoflurane组PDTC+Sevoflurane组培养的小胶质细胞盒内释放21%O2,5%CO2,4.1%七氟烷的气体,用气体分析仪持续监测各组的浓度。应用Iba-1的免疫荧光染色法对小鼠小胶质细胞进行纯度鉴定。分别在于给七氟烷后2 h、4 h和6 h时采用免疫印迹分析技术检测两组小胶质细胞IL-6和TNF-α的表达水平和NF-κB的活性。PDTC+Sevoflurane组在给七氟烷前一小时给予PDTC,采用ELISA技术和免疫印迹分析技术检测各组小胶质细胞IL-6和TNF-α的浓度和NF-κB的表达。结果:免疫印迹显示七氟烷组细胞中IL-6、TNF-α水平和NF-κB的激活水平升高;PDTC降低了七氟烷作用后核内NF-κB的表达,减弱了IL-6和TNF-α水平的升高作用。结论:七氟烷可通过激活NF-κB信号通路,进一步激活培养的小鼠小胶质细胞中炎症因子的表达。     

绞股蓝多糖对糖尿病肾病大鼠肾功能的保护作用及其机制研究

目的:探讨绞股蓝多糖对糖尿病大鼠肾功能的影响及其保护的可能机制。方法:采用大鼠高脂高糖饲料结合腹腔注射链脲佐菌素建立2型糖尿病模型。灌胃给予绞股蓝多糖8周后,测定大鼠空腹血糖(FBG),血清肌酐、血清尿素氮、24 h尿量及尿蛋白等指标;组织病理切片观察肾形态,肾小球体积;Western blot法检测肾皮质NF-κB的表达。结果:绞股蓝多糖(GPS)可剂量依赖性的降低DN组的FBG、血肌酐、24 h尿量和尿蛋白;但血尿素氮模型组与GPS组无统计学差异;中、高剂量GPS治疗后,肾脏指数较模型组明显降低(P0.05);与模型组(149.8±12.2%)比较,高剂量GPS可抑制肾小球体积肥大至108.9±9.6%。进一步研究发现,GPS可剂量依赖性的降低DN组肾脏NF-κB蛋白的表达,高、中剂量组水平与模型组相比有统计学差异(P0.05)。结论:绞股蓝多糖对实验性糖尿病肾病具有保护作用,其机制可能与抑制肾炎症相关通路NF-κB的表达有关。     

NF-κB抑制剂通过逆转LPS诱导的BMPRⅡ下调改善肺血管重构

动脉性肺动脉高压(pulmonary arterial hypertension, PAH)的发生、发展与骨形态发生蛋白受体II型(bone morphogenetic protein receptor type II, BMPRII)编码基因的遗传突变和核因子κB (nuclear factorκB, NF-κB)通路介导的炎症反应密切相关。本文旨在研究NF-κB通路抑制剂对脂多糖(lipopolysaccharide, LPS)诱导的肺动脉内皮细胞损伤的作用。用1μg/mL的LPS处理人肺动脉内皮细胞,用免疫印迹和qPCR检测BMPRII和白介素8 (interleukin-8, IL-8)的表达水平。腹腔注射野百合碱(monocrotaline, MCT)建立大鼠PAH模型,用免疫荧光染色法检测肺动脉内皮细胞BMPRII和IL-8的表达情况,检测模型大鼠心脏血流动力学变化和肺血管重构情况。结果显示,LPS可引起人肺动脉内皮细胞BMPRII的表达下调和IL-8的表达上调,NF-κB抑制剂BAY11-7082 (10μmol/L)可逆转LPS的作用。在MCT-PAH大鼠模型中,肺动脉内皮细胞BMPRII表达下调,IL-8表达上调,右心室/(左心室+室间隔)重量比值[weight ratio of right ventricle to left ventricle plus septum, RV/(LV+S)]和右心室收缩压(right ventricular systolic pressure, RVSP)显著升高,心输出量(cardiac output, CO)和三尖瓣环收缩期位移(tricuspid annular plane systolic excursion, TAPSE)明显降低,肺血管壁明显增厚,连续21天腹腔注射BAY11-7082 (5 mg/kg)可逆转上述变化。以上结果提示,LPS通过NF-κB信号通路下调BMPRII的表达水平,促进PAH的发生、发展,因此NF-κB信号通路可作为PAH潜在治疗靶点。     

芹菜素对缺血再灌注大鼠心肌核转录因子-κB和ICAM-1表达的影响

目的研究芹菜素对缺血再灌注大鼠心肌核转录因子-κB(NF-κB)和粘附分子ICAM-1表达的影响,探讨其抗心肌缺血再灌注损伤的可能机制。方法将Wistar大鼠随机分为6组:假手术组、缺血再灌注组(IR组)、溶剂对照组、维拉帕米阳性对照组、芹菜素低、高剂量用药组。采用结扎左冠状动脉前降支制作缺血再灌注模型,心肌缺血30 min,再灌注2 h。免疫组化染色检测心肌组织的NF-κB和ICAM-1的表达.结果芹菜素组可降低心肌组织NF-κB和ICAM-1的表达,与IR组比较有显著差异(P0.05)。结论芹菜素对缺血再灌注心肌的保护作用可能与其减少心肌缺血再灌注后NF-κB和ICAM-的激活有关。     

抑制NF-κB信号通路对缺氧诱导小鼠海马神经元HT22细胞损伤的影响

本研究试图从细胞衰老和凋亡方面,探讨NF-κB信号通路抑制剂PDTC对缺氧诱导的小鼠海马神经细胞HT22的影响。实验研究以小鼠海马神经元细胞HT22细胞为细胞模型,设置4组,分别是Control组、缺氧组、缺氧+PDTC组、PDTC组(PDTC, NF-κB信号通路抑制剂)。用CCK-8法检测细胞存活率;hochest33258染色观察细胞凋亡情况;Elisa检查细胞Caspase-3水平;Western blotting检测衰老相关蛋白P16和P21的表达。CCK-8结果发现,与缺氧组比较,缺氧+PDTC组中HT22细胞数存活率明显升高;hochest33258染色结果示:与缺氧组比较,缺氧+PDTC组中hochest33258染色阳性HT22细胞数目显著降低;Elisa结果显示,缺氧+PDTC组HT22细胞中Caspase-3水平与缺氧组比较显著降低。Western blotting结果显示:与缺氧组比较,缺氧+PDTC组HT22细胞中衰老相关蛋白P16和P21表达明显降低。本研究初步结果表明,运用PDTC抑制NF-κB信号通路能有效拮抗缺氧对小鼠海马细胞HT22细胞的损伤作用。     

NF-κB介导非对称性二甲基精氨酸上调大鼠主动脉诱导型一氧化氮合酶的表达

目的探讨非对称性二甲基精氨酸(ADMA)上调大鼠主动脉诱导型一氧化氮合酶(iNOS)的表达是否是通过激活NF-κB实现的。方法 Wistar大鼠50只随机分为四组:①对照组(n=10):标准饲料喂养。②H组(n=12):高脂饲料喂养。③A+H组(n=14):予ADMA[0.2mg/kgd]灌胃,高脂饲料喂养。④P+A+H组(n=14):予吡咯烷二硫代氨基甲酸盐(pyrrolidine dithiocarbamate,PDTC)[40mg/kgd]腹腔注射、ADMA[0.2mg/kgd]灌胃、高脂饲料喂养。对照组、H组予等体积的生理盐水灌胃及腹腔注射、A+H组给予等体积的生理盐水腹腔注射。18周后麻醉大鼠、取主动脉。以实时荧光定量PCR和Westen blotting分别检测iNOS mRNA和蛋白表达,以电泳迁移率变动分析(EMSA)和增强化学发光法(ECL)检测NF-κB活性。结果①A+H组iNOS mRNA和蛋白表达量较对照组和H组增加(P<0.05),P+A+H组较A+H组iNOSmRNA和蛋白表达量降低(P<0.05)。②A+H组NF-κB活性较对照组和H组显著升高(P<0.05),P+A+H组NF-κB活性较A+H组明显降低(P<0.05)③相关分析:NF-κB活性与iNOS mRNA和蛋白表达量呈正相关(相关系数r分别为0.854、0.876,P<0.05)。结论 ADMA可能通过激活NF-κB途径上调iNOS表达,从而促进动脉粥样硬化的发生发展。     

C3a Receptor Antagonist Ameliorates Inflammatory and Fibrotic Signals in Type 2 Diabetic Nephropathy by Suppressing the Activation of TGF-β/smad3 and IKBα Pathway

Objective

Diabetic nephropathy (DN) is a serious complication for patients with diabetes mellitus (DM). Emerging evidence suggests that complement C3a is involved in the progression of DN. The aim of this study was to investigate the effect of C3a Receptor Agonist (C3aRA) on DN and its potential mechanism of action in rats with type 2 diabetes mellitus (T2DM).

Methods

T2DM was induced in SD rats by a high fat diet (HFD) plus repeated low dose streptozocin (STZ) injections. T2DM rats were treated with vehicle or C3aRA for 8 weeks. Biochemical analysis, HE and PAS stains were performed to evaluate the renal function and pathological changes. Human renal glomerular endothelial cells (HRGECs) were cultured and treated with normal glucose (NG), high glucose (HG), HG+C3a, HG+C3a+C3aRA and HG+C3a+BAY-11-7082 (p-IKBα Inhibitor) or SIS3 (Smad3 Inhibitor), respectively. Real-time PCR, immunofluorescent staining and western blot were performed to detect the mRNA and protein levels, respectively.

Results

T2DM rats showed worse renal morphology and impaired renal function compared with control rats, including elevated levels of serum creatinine (CREA), blood urea nitrogen (BUN) and urine albumin excretion (UACR), as well as increased levels of C3a, C3aR, IL-6, p-IKBα, collagen I, TGF-β and p-Smad3 in the kidney of T2DM rats and C3a-treated HRGECs. In contrast, C3aRA treatment improved renal function and morphology, reduced CREA, UACR and the intensity of PAS and collagen I staining in the kidney of T2DM rats, and decreased C3a, p-IKBα, IL-6, TGF-β, p-Smad3 and collagen I expressions in HRGECs and T2DM rats.

Conclusion

C3a mediated pro-inflammatory and pro-fibrotic responses and aggravated renal injury in T2DM rats. C3aRA ameliorated T2DN by inhibiting IKBα phosphorylation and cytokine release, and also TGF-β/Smad3 signaling and ECM deposition. Therefore, complement C3a receptor is a potential therapeutic target for DN.     

Manganese Supplementation Reduces High Glucose-induced Monocyte Adhesion to Endothelial Cells and Endothelial Dysfunction in Zucker Diabetic Fatty Rats

Endothelial dysfunction is a hallmark of increased vascular inflammation, dyslipidemia, and the development of atherosclerosis in diabetes. Previous studies have reported lower levels of Mn²⁺ in the plasma and lymphocytes of diabetic patients and in the heart and aortic tissue of patients with atherosclerosis. This study examines the hypothesis that Mn²⁺ supplementation can reduce the markers/risk factors of endothelial dysfunction in type 2 diabetes. Human umbilical vein endothelial cells (HUVECs) were cultured with or without Mn²⁺ supplementation and then exposed to high glucose (HG, 25 mm) to mimic diabetic conditions. Mn²⁺ supplementation caused a reduction in monocyte adhesion to HUVECs treated with HG or MCP-1. Mn²⁺ also inhibited ROS levels, MCP-1 secretion, and ICAM-1 up-regulation in HUVECs treated with HG. Silencing studies using siRNA against MnSOD showed that similar results were observed in MnSOD knockdown HUVECs following Mn²⁺ supplementation, suggesting that the effect of manganese on monocyte adhesion to endothelial cells is mediated by ROS and ICAM-1, but not MnSOD. To validate the relevance of our findings in vivo, Zucker diabetic fatty rats were gavaged daily with water (placebo) or MnCl₂ (16 mg/kg of body weight) for 7 weeks. When compared with placebo, Mn²⁺-supplemented rats showed lower blood levels of ICAM-1 (17%, p < 0.04), cholesterol (25%, p < 0.05), and MCP-1 (28%, p = 0.25). These in vitro and in vivo studies demonstrate that Mn²⁺ supplementation can down-regulate ICAM-1 expression and ROS independently of MnSOD, leading to a decrease in monocyte adhesion to endothelial cells, and therefore can lower the risk of endothelial dysfunction in diabetes.     

Astragaloside IV ameliorates renal injury in streptozotocin-induced diabetic rats through inhibiting NF-κB-mediated inflammatory genes expression

Accumulating evidence suggests that inflammatory processes are involved in the development of diabetic nephropathy (DN). However, there are no effective interventions for inflammation in the diabetic kidneys. Here, we tested the hypothesis that Astragaloside IV(AS-IV), a novel saponin purified from Astragalus membranaceus (Fisch) Bge, ameliorates DN in streptozotocin (STZ)-induced diabetic rats through anti-inflammatory mechanisms. Diabetes was induced with STZ (65 mg/kg) by intraperitoneal injection in rats. Two weeks after STZ injection, rats were divided into three groups (n = 8/each group), namely, diabetic rats, diabetic rats treated with AS-IV at 5 and 10 mg kg^?1 d^?1, p.o., for 8 weeks. The normal rats were chosen as nondiabetic control group (n = 8). The rats were sacrificed 10 weeks after induction of diabetes. AS-IV ameliorated albuminuria, renal histopathology and podocyte foot process effacement in diabetic rats. Renal NF-κB activity, as wells as protein and mRNA expression were increased in diabetic kidneys, accompanied by an increase in mRNA expression and protein content of TNF-α, MCP-1 and ICAM-1 in kidney tissues. The α₁-chain type IV collagen mRNA was elevated in the kidneys of diabetic rats. All of these abnormalities were partially restored by AS-IV. AS-IV also decreased the serum levels of TNF-α, MCP-1 and ICAM-1 in diabetic rats. These findings suggest that AS-IV, a novel anti-inflammatory agent, attenuated DN in rats through inhibiting NF-κB mediated inflammatory genes expression.     

FPS-ZM1 and valsartan combination protects better against glomerular filtration barrier damage in streptozotocin-induced diabetic rats

Despite the effectiveness of renin-angiotensin blockade in retarding diabetic nephropathy progression, a considerable number of patients still develop end-stage renal disease. The present investigation aims to evaluate the protective potential of FPS-ZM1, a selective inhibitor of receptor for advanced glycation end products (RAGE), alone and in combination with valsartan, an angiotensin receptor blocker, against glomerular injury parameters in streptozotocin-induced diabetic rats. FPS-ZM1 at 1 mg/kg (i.p.), valsartan at 100 mg/kg (p.o.), and their combination were administered for 4 weeks, starting 2 months after diabetes induction in rats. Tests for kidney function, glomerular filtration barrier, and podocyte slit diaphragm integrities were performed. Combined FPS-ZM1/valsartan attenuated diabetes-induced elevations in renal levels of RAGE and phosphorylated NF-κB p65 subunit. It ameliorated glomerular injury due to diabetes by increasing glomerular nephrin and synaptopodin expressions, mitigating renal integrin-linked kinase (ILK) levels, and lowering urinary albumin, collagen type IV, and podocin excretions. FPS-ZM1 also improved renal function as demonstrated by decreasing levels of serum cystatin C. Additionally, the combination also alleviated indices of renal inflammation as revealed by decreased renal monocyte chemoattractant protein 1 (MCP-1) and chemokine (C-X-C motif) ligand 12 (CXCL12) expressions, F4/80-positive macrophages, glomerular TUNEL-positive cells, and urinary alpha-1-acid glycoprotein (AGP) levels. These findings underline the benefits of FPS-ZM1 added to valsartan in alleviating renal glomerular injury evoked by diabetes in streptozotocin rats and suggest FPS-ZM1 as a new potential adjunct to the conventional renin-angiotensin blockade.     

The CREB/KMT5A complex regulates PTP1B to modulate high glucose-induced endothelial inflammatory factor levels in diabetic nephropathy

Diabetic nephropathy (DN) is the primary microvascular complication of diabetes mellitus and may result in end-stage renal disease. The overproduction of various inflammatory factors is involved in the pathogenesis of DN. Protein tyrosine phosphatase 1B (PTP1B) modulates the expression of a series of cytokines and nuclear factor kappa B (NF-κB) activity. cAMP response element-binding protein (CREB) and lysine methyltransferase 5A (KMT5A) have been reported to participate in the maintenance of a healthy endothelium. In the present study, we hypothesise that CREB associates with KMT5A to modulate PTP1B expression, thus contributing to high glucose-mediated glomerular endothelial inflammation. Our analyses revealed that plasma inflammatory factor levels, glomerular endothelial p65 phosphorylation and PTP1B expression were increased in DN patients and rats. In vitro, high glucose increased endothelial inflammatory factor levels and p65 phosphorylation by augmenting PTP1B expression in human umbilical vein endothelial cells (HUVECs). Moreover, high glucose decreased CREB and KMT5A expression. CREB overexpression and KMT5A overexpression both inhibited high glucose-induced PTP1B expression, p65 phosphorylation and endothelial inflammatory factor levels. si-CREB- and sh-KMT5A-induced p65 phosphorylation and endothelial inflammatory factor levels were reversed by si-PTP1B. Furthermore, CREB was associated with KMT5A. Mechanistic research indicated that CREB and histone H4 lysine 20 methylation (H4K20me1, a downstream target of KMT5A) occupy the PTP1B promoter region. sh-KMT5A augmented PTP1B promoter activity and activated the positive effect of si-CREB on PTP1B promoter activity. Our in vivo study demonstrated that CREB and KMT5A were downregulated in glomerular endothelial cells of DN patients and rats. In conclusion, CREB associates with KMT5A to promote PTP1B expression in vascular endothelial cells, thus contributing to hyperglycemia-induced inflammatory factor levels in DN patients and rats.Subject terms: Diabetes complications, Type 2 diabetes     

胰岛素对大鼠急性肺损伤的肺血管内皮细胞核因子-kB和细胞间粘附分子-1表达的影响

目的 探讨胰岛素对急性肺损伤（ALI）大鼠肺血管内皮细胞核因子-kB（NF-kB）和细胞间粘附分子-1（ICAM-1）表达的影响。方法24只健康雄性SD大鼠（190-210g）,随机分为正常对照组、ALI模型组、胰岛素干预组。观察肺组织病理形态,采用原位杂交技术半定量法和免疫组织化学染色检测肺血管内皮细胞的细胞间粘附分子-1（ICAM-1）mRNA和核因子-kB（NF-kB）蛋白的表达。结果（1）肺病理组织学结果显示胰岛素干预组肺病变局限且程度减轻;（2）ALI模型组肺血管内皮细胞ICAM-1mRNA表达（O．456±0．018）和NF-kB核染色阳性细胞百分比（0．542±0．009）与正常对照组（o．274土0．014,0．308±0．017）比较显著升高（均P〈0．05）;（3）胰岛素干预组ICAM-1mRNA表达（0．357±0．024）和NF-kB核染色阳性细胞百分比（0．427±0．018）比模型组明显减低（均P〈0．05）,但与正常对照组比较仍较高（均P＜0．05）。结论ICAM-1和NF-kB在ALI显著增加,胰岛素可以抑制NF-kB和ICAM-1mRNA的表达,可能是其对抗ALI的作用机制之一。     

Hemolysate-induced Expression of Intercellular Adhesion Molecule-1 and Monocyte Chemoattractant Protein-1 Expression in Cultured Brain Microvascular Endothelial Cells via Through ROS-dependent NF-κB Pathways

In order to determine the possible effects of hemolysate on brain microvascular endothelial cells (BMECs), we examined the effects of hemolysate on the expression of intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1), generation of reactive oxygen species (ROS), and NF-κB activation in rat BMECs. Hemolysate induced the expression of ICAM-1 and MCP-1 in endothelial cells. In addition, hemolysate stimulated nuclear translocation of the p65 subunit of NF-κB, and NF-κB DNA-binding activity in BMECs. Furthermore, hemolysate increased ROS generation, and hemolysate-induced ICAM-1and MCP-1 expression and NF-κB activation were abrogated in the presence of the direct scavenger of ROS. Taken together, our results indicate that hemolysate can induce inflammatory responses that increase expression of ICAM-1 and MCP-1, through ROS-dependent NF-κB activation in BMECs.     

Thioredoxin-interacting protein mediates NALP3 inflammasome activation in podocytes during diabetic nephropathy

Numerous studies have shown that the NALP3 inflammasome plays an important role in various immune and inflammatory diseases. However, whether the NALP3 inflammasome is involved in the pathogenesis of diabetic nephropathy (DN) is unclear. In our study, we confirmed that high glucose (HG) concentrations induced NALP3 inflammasome activation both in vivo and in vitro. Blocking NALP3 inflammasome activation by NALP3/ASC shRNA and caspase-1 inhibition prevented IL-1β production and eventually attenuated podocyte and glomerular injury under HG conditions. We also found that thioredoxin (TRX)-interacting protein (TXNIP), which is a pro-oxidative stress and pro-inflammatory factor, activated NALP3 inflammasome by interacting with NALP3 in HG-exposed podocytes. Knocking down TXNIP impeded NALP3 inflammasome activation and alleviated podocyte injury caused by HG. In summary, the NALP3 inflammasome mediates podocyte and glomerular injury in DN, moreover, TXNIP participates in the formation and activation of the NALP3 inflammasome in podocytes during DN, which represents a novel mechanism of podocyte and glomerular injury under diabetic conditions.     

Influence of myrcene on inflammation,matrix accumulation in the kidney tissues of streptozotocin-induced diabetic rat

There is only limited literature studies on the activities of inflammation and matrix accumulation in the renal tissues of rats induced with diabetes through Streptozotocin. The present the investigation involves the examination of the protective actions of Myrcene (MYN), a monoterpene on the oxidative stress, inflammation, and matrix accumulation. For this purpose an experimental setup was created which involves injecting MYN 50 mg/kg for about 45 days in the STZ diabetic rats. Modifications in the enzymes, collagens, growth factor B₁ and Kappa factor P65 were identified and tracked. The levels of the inflammatory markers like TF-α1, ICAM-1, VCAM-1, MCP-1 were tracked and noted. The current experimental results showed an alteration in the glucose metabolism and enhanced condition. Also an increased level of TGF-β-1 and Nuclear factor-kB expression was seen in the renal tissues. MYN was found to reduce glucose oxidative stress and exhibit an anti-inflammatory effect via inhibiting NF-kB signalling. The conclusion of the current study reveals that MYN regulates the inflammatory activities and matrix accumulation by inhibiting the activities of inflammatory cytokine, pro-inflammatory signalling.     

Angiotensin II Upregulates Endothelial Lipase Expression via the NF-Kappa B and MAPK Signaling Pathways

Background

Angiotensin II (AngII) participates in endothelial damage and inflammation, and accelerates atherosclerosis. Endothelial lipase (EL) is involved in the metabolism and clearance of high density lipoproteins (HDL), the serum levels of which correlate negatively with the onset of cardiovascular diseases including atherosclerosis. However, the relationship between AngII and EL is not yet fully understood. In this study, we investigated the effects of AngII on the expression of EL and the signaling pathways that mediate its effects in human umbilical vein endothelial cells (HUVECs).

Methods and Findings

HUVECs were cultured in vitro with different treatments as follows: 1) The control group without any treatment; 2) AngII treatment for 0 h, 4 h, 8 h, 12 h and 24 h; 3) NF-κB activation inhibitor pyrrolidine dithiocarbamate (PDTC) pretreatment for 1 h before AngII treatment; and 4) mitogen-activated protein kinase (MAPK) p38 inhibitor (SB203580) pretreatment for 1 h before AngII treatment. EL levels in each group were detected by immunocytochemical staining and western blotting. HUVECs proliferation was detected by MTT and proliferating cell nuclear antigen (PCNA) immunofluorescence staining. NF-kappa B (NF-κB) p65, MAPK p38, c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and phosphorylated extracellular signal-regulated kinase (p-ERK) expression levels were assayed by western blotting. The results showed that the protein levels of EL, NF-κB p65, MAPK p38, JNK, and p-ERK protein levels, in addition to the proliferation of HUVECs, were increased by AngII. Both the NF-kB inhibitor (PDTC) and the MAPK p38 inhibitor (SB203580) partially inhibited the effects of AngII on EL expression.

Conclusion

AngII may upregulate EL protein expression via the NF-κB and MAPK signaling pathways.