Reactive Oxygen Species Independent Cytotoxicity Induced by Radiocontrast Agents in Tubular Cells (LLC-PK1 and MDCK)

Purpose. Radiocontrast agents (RAs) cause renal tubular damage by hemodynamic imbalance, which could cause hypoxic stimulus and direct cytotoxicity. However, reactive oxygen species (ROS) could be an important factor in RAs' direct cytotoxicity. This study investigated the involvement of ROS in deleterious effects produced by RAs on normoxic and hypoxic renal tubular cells. Materials and Methods. LLC-PK1 and MDCK were exposed to diatrizoate and ioxaglate in normoxic and hypoxic conditions. Apoptotic and necrotic cell death were assessed by acridine orange/ethidium bromide and annexin V methods. Hydrogen peroxide, superoxide anion, and malondialdehyde levels were analyzed by, respectively, 2′,7′-dichlorofluorescein, luminal, and thiobarbituric acid. Antioxidant agents were used to prevent cellular RAs damage. Results. Diatrizoate and ioxaglate decreased cellular viability in both cells, and this effect was enhanced by hypoxic conditions. Diatrizoate induced more injury than ioxaglate to both cell lines. LLC-PK1 underwent necrosis, while MDCK cells underwent apoptosis when exposed to diatrizoate. These results could not be attributed to an increase in osmolality. RAs did not increase hydrogen peroxide, superoxide anion or malondialdehyde levels in both cells. Additionally, N-acetyl-L-cysteine (NAC), ascorbic acid, α-tocopherol, glutathione, β-carotene, allopurinol, cimetidine, and citric acid did not protect cells against RAs damage. Surprising, NAC increased the cellular damage induced by ioxaglate in the both cell lines. Conclusion. The present study shows that RAs induce damage in cultured tubular cells, especially in hypoxic conditions. ROS were not involved in the observed RAs' cytotoxicity, and NAC increased ioxaglate-induced tubular damage.     

P选择素在油酸诱导急性肺损伤大鼠肺组织中的表达及N-乙酰半胱氨酸对其影响的研究

目的探讨油酸(Olic Acid,OA)诱导的急性肺损伤(Acute Lung Injury,ALI)大鼠肺血管内皮P-选择素(P-selectin,Ps)表达的变化及N-乙酰半胱氨酸(N-acetylcysteine,NAC)的保护作用。方法成年雄性SD大鼠42只按随机化原则分为3组:油酸组(OA组n=18);对照组(n=6); N-乙酰半胱氨酸 油酸组(NAC组,n=18);按注射油酸后不同时间随机分为3组,1小时、3小时、6 小时组,行P选择素免疫组织化学染色及光密度测值,并检测肺组织匀浆髓过氧化物酶(MPO)、超氧化物岐化酶(SOD)、丙二醛(MDA)。结果Ps在OA诱导的ALI大鼠肺血管内皮1小时出现表达,3 小时表达达高峰,持续表达6小时(P<0．01);OA组各时点MPO、MDA升高,与对照组比较差异均有显著性(P<0．01),NAC组各时点MPO、MDA均低于相应时点OA组(P<0．01),但均高于对照组(P<0．01);OA组SOD明显降低,与对照组比较差异明显(P<0．01),NAC组各时点SOD均高于相应时点OA组(P<0．01),但均低于对照组(P<0．01);Ps表达与MPO、MDA成正相关(r=0．793, 0．886;P<0．01)。结论Ps在OA诱导的ALI大鼠肺血管内皮表达1小时升高,3小时表达达高峰, 持续表达6小时;NAC通过清除氧自由基抑制Ps在肺血管内皮的表达,对OA诱导ALI大鼠起保护作用。     

Hepatoprotective effects of antioxidants in chronic hepatitis C

We have read with interest the paper published in issue 2, volume 16 of World Journal of Gastroenterology 2010 by Nakamura et al, demonstrating that the antioxidant resveratrol (RVT) enhances hepatitis C virus (HCV) replication, consequently, they conclude that RVT is not a suitable antioxidant therapy for HCV chronic infection. The data raise some concern regarding the use of complementary and alternative medicine since the most frequent supplements taken by these patients are antioxidants or agents that m...     

Caspase-3在大鼠挫伤性视网膜病变中的表达及NAC对其表达的影响

目的 研究凋亡相关基因Caspase-3在大鼠挫伤性视网膜病变中的表达及N-乙酰-L-半胱氨酸(NAC)对其表达的影响.方法 自由落体法制作视网膜挫伤模型.将SD大鼠随机分为正常组、挫伤组及N-乙酰半胱氨酸(NAC)治疗组.每组按挫伤后不同时间段分为1 d、3d、7d及14d组,HE染色光镜观察视网膜组织变化,应用SABC免疫组织化学法检测大鼠视网膜挫伤后视网膜组织caspase-3蛋白表达的变化.结果 正常组视网膜组织Caspase-3不表达.视网膜挫伤后1 d时caspase-3蛋白表达开始增多,3 d强阳性表达,7 d表达有所减少,14 d时表达进一步减少.NAC治疗组各观察指标变化趋势大体与挫伤组相似,但表达明显较挫伤组弱,两组相比较,于挫伤后1、3和7 d时差异有统计学意义(P<0.05).结论 凋亡相关基因Caspase-3参与了挫伤性视网膜病变的形成,NAC可以抑制Caspase-3蛋白的表达;NAC在挫伤性视网膜病变中可能通过抑制细胞凋亡而达到其保护作用.     

乙酰半胱氨酸水平与单胺类物质所致神经元损害的关系

目的：探讨抗氧化剂在高浓度单胺类物质（MA）条件下对神经细胞的保护作用。方法：应用体外培养神经细胞，分别采用琼脂糖凝胶电泳、原位末端标记及流式细胞仪定性定量分析5、10、20μmol/L乙酰半胱氨酸（NAC）对300μmol/L多巴胺、去甲肾上腺素、5－羟色胺处理24h后的神经细胞凋亡的影响。结果：多巴胺、去甲肾上腺素及5－羟色胺诱导神经细胞的死亡具有凋亡特征。乙酰半胱氨酸对神经细胞有保护作用，且在5－10μmol/L范围内有剂量效应，20μmol/L较5－10μmol/L乙酰半胱氨酸对神经细胞的保护作用明显减弱。结论：高浓度单胺类物质诱导神经细胞凋亡，适当浓度乙酰半胱氨酸对神经细胞有保护作用。     

N-乙酰半胱胺酸对大鼠挫伤性视网膜病变的保护作用

目的:研究凋亡相关基因Bcl-2/Bax在大鼠挫伤性视网膜病变中的表达,以及N-乙酰半胱氨酸(N-acetyl-cysteine,NAC)对其表达的影响。方法:自由落体法制作视网膜挫伤模型。将SD大鼠随机分为正常组6只、模型组24只及治疗组24只。每组按挫伤后不同时间段分为1,3,7,14d,每个观察时段6只大鼠。HE染色光镜观察视网膜组织学变化,应用SABC免疫组织化学法检测大鼠视网膜挫伤后视网膜组织Bcl-2/Bax蛋白表达的变化。结果:挫伤性视网膜病变的损害主要集中在神经纤维层。挫伤后视网膜组织水肿,细胞紊乱,胞浆空泡样变,视网膜组织变薄,细胞丢失。NAC治疗组视网膜水肿程度有所改善,细胞紊乱,胞浆空泡样变,细胞丢失有所恢复。正常组及NAC治疗组视网膜组织Bax均未见表达,Bcl-2低表达。视网膜挫伤后1d时Bax表达开始增多,3d强阳性表达,7d表达有所减少,14d时表达进一步减少。Bcl-2低表达,未见明显变化。NAC治疗组各观察指标变化趋势基本与视网膜挫伤组相似,但表达明显减弱,两组相比较,于挫伤后1,3d和7d时差异有统计学意义(P<0.05)。Bcl-2在各时段的表达均较模型组有所增强,两组相比较,于挫伤后1,3d和7d时差异有统计学意义(P<0.01)。结论:在大鼠挫伤性视网膜病变中,NAC能够改善视网膜组织病理学损害并通过调节Bcl-2/Bax蛋白表达而抑制细胞凋亡,对视网膜挫伤具有保护作用。     

High mobility group box-1-toll-like receptor 4-phosphatidylinositol 3-kinase/protein kinase B-mediated generation of matrix metalloproteinase-9 in the dorsal root ganglion promotes chemotherapy-induced peripheral neuropathy

Chemotherapy-induced peripheral neuropathy (CIPN) is a significant side effect of chemotherapeutics. The mechanisms of CIPN remain substantially unidentified, although inflammation-induced peripheral sensitization has been indicated as an important factor. Here, we aimed to illustrate the role of the matrix metalloproteinase (MMP)-9-related signaling pathway in the process of CIPN. Oxaliplatin (L-OHP) was administered to mice to establish the CIPN model. Gelatin zymography was used to measure MMP-9/2 activities. Western blotting and immunohistochemistry were used to measure the expression of high-mobility group box-1 (HMGB-1), calcitonin gene-related peptide and ionized calcium-binding adapter molecule 1. Mechanical withdrawal was measured by von Frey hairs testing. Raw 264.7 cells and SH-SY5Y cells were cultured to investigate cell signaling in vitro. Here, we report that L-OHP-induced mechanical pain in mice with significant MMP-9/2 activation in dorsal root ganglion (DRG) neurons. MMP-9 inhibition or knockout alleviated the occurrence of CIPN directly. MMP-9/2 were released from macrophages and neurons in the DRG via the HMGB-1-toll-like receptor 4 (TLR4)-phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) axis, because MMP-9/2 activities could be reduced by macrophage scavengers or PI3Kγ knockout in CIPN mice. The in vitro data revealed that induced MMP-9 activity by recombinant HMGB-1 could be abolished by TLR4, PI3K or Akt inhibitors. Finally, it was shown that N-acetyl-cysteine (NAC) could reduce MMP-9/2 activities and attenuate CIPN effectively and safely. The HMGB-1-TLR4-PI3K/Akt-MMP-9 axis is involved in the crosstalk between macrophages and neurons in the pathological process of CIPN in mice. Direct inhibition of MMP-9 by NAC may be a potential therapeutic regimen for CIPN treatment.     

The role of N-acetyl-cysteine (NAC) orally daily on the sperm parameters and serum hormones in idiopathic infertile men: A systematic review and meta-analysis of randomised controlled trials

The meta-analysis was performed to access the role of N-acetyl-cysteine (NAC) orally daily on the sperm parameters and serum hormones in idiopathic infertile men. Randomised controlled trials (RCTs) were retrieved using PubMed, EMBASE and Cochrane register databases. The references of included studies were also searched. Finally, three articles including 431 infertile men were analysed. The results indicated that the NAC group had a considerable improvement in sperm concentration (mean difference [MD], 3.80; p < .00001), ejaculate volume (MD, 0.69; p = .002), sperm motility (MD, 4.69; p < .0001) and normal morphology (MD, 1.68; p = .0002) compared with the placebo group. However, in terms of serum hormones, the NAC group did not show significant difference in increasing the serum levels of testosterone (MD, 1.35; p = .21), luteinising hormone (MD, 0.82; p = .40), follicle-stimulating hormone (MD, −7.48; p = .29) and prolactin (MD, −0.34; p = .32) compared with the placebo group. In conclusion, NAC orally daily produced a greater improvement in sperm concentration, ejaculate volume, sperm motility and normal morphology for idiopathic infertile men, whereas no significant influence in serum hormones, which required more high-quality RCTs with sufficient sample sizes and statistics to prove.