RBC aggregation,deformation and adhesion to endothelium:Role of nitric oxide derived from L-Arginine and sodium nitroprusside

Red blood cells(RBCs)are the most abundant human blood cells.RBC aggregation and deformation strongly determine blood viscosity which impacts hemorheology and microcirculation.In turn,RBC properties depend on di®erent endogenous and exogenous factors.One such factor is nitric oxide(NO),which is mainly produced by endothelial cells(EC)from L-arginine amino acid in the circulatory system.Since the mechanisms of the RBC-endothelium interplay are not clear up to date and considering its possible clinical importance,the aims of this study are to investigate in vitro:(1)The effect of L-arginine induced NO on RBC aggregation and adhesion to endothelium;(2)the NO e®ect on RBC aggregation and deformation induced by L-arginine and sodium nitroprusside without the presence of endothelium in the samples.The RBC aggregation and adhesion to a monolayer of EC were studied using optical tweezers(OT).The RBC deformability and aggregation without endothelium in the samples were studied using the flow chamber method and Myrenne aggregometer.We confirmed that NO increases deformability and decreases aggregation of RBCs.We showed that the soluble guanylate cyclase pathway appears to be the only NO signaling pathway involved.In the samples with the endothelium,the "bell-shaped"dependence of RBC aggregation force on L-arginine concentration was observed,which improves our knowledge about the process of NO production by endothelium.Additionally,data related to L-arginine accumulation by endothelium were obtained:Necessity of the presence of extracellular L-arginine stated by other authors was put under question.In our study,NO decreased the RBC-endothelium adhesion,however,the tendency appeared to be weak and was not confirmed in another set of experiments.To our knowledge,this is the first attempt to measure the forces of RBC adhesion to endothelium monolayer with OT.

L-Arginine Supplementation Mitigates Dichlorvos-Induced Haematocardiotoxicity, and Oxidative Stress in Male Wistar Rats

Due to its toxicity, dichlorvos—a common organophosphate pesticide—poses significant risks to human health. This study utilized male Wistar rats to explore the potential protective effects of L-arginine supplementation against dichlorvos-induced toxicity, focusing on cardiotoxicity, haematotoxicity and oxidative stress. The rats were divided into four groups: Control, L-arginine (L), Dichlorvos (D), and L-arginine + Dichlorvos (L + D). Dichlorvos was administered to the D group, L-arginine (100 mg/kg) to the L group, and both L-arginine and dichlorvos to the L + D group. The study evaluated various parameters, including cardiovascular, oxidative stress markers, and haematological indices. Significant changes in haematological parameters such as haemoglobin (Hb), haematocrit (HCT), and red blood cell count (RBC) indicated haematotoxicity after dichlorvos administration. Additionally, elevated cardiac markers, including lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB), suggested cardiotoxic effects. Exposure to dichlorvos also resulted in decreased antioxidant enzyme levels and increased oxidative stress indicators like malondialdehyde (MDA). Remarkably, L-arginine supplementation mitigated the damage caused by dichlorvos. It normalized the altered haematological parameters, demonstrating its protective effect against haematotoxicity. The rise in cardiac markers was reduced with L-arginine supplementation, indicating protection against cardiotoxicity. Moreover, L-arginine significantly decreased oxidative stress, as evidenced by lower MDA levels and restored antioxidant enzyme activity. In conclusion, L-arginine supplementation in male Wistar rats showed promising protective effects against dichlorvos-induced cardiotoxicity, haematotoxicity and oxidative stress. This suggests that L-arginine may offer a beneficial intervention to mitigate the adverse effects of dichlorvos on blood and heart health, paving the way for potential treatments for pesticide poisoning.

L-Arginine及衰老对大鼠阴茎组织中NO、ET-1的影响

目的 探讨喂养左旋精氨酸 (L Arginine)及衰老对大鼠阴茎组织中“NO cGMP通路”及ET 1的影响及意义。方法 将不同月龄 ( 2、8、16、2 4月 )大鼠随机分为对照组与实验组 (喂养L Arginine) ,进行了以下研究 :①阴茎组织中一氧化氮(nitricoxide ,NO)、环磷酸鸟苷 (cGMP)含量测定 ;②阴茎组织一氧化氮合酶 (nitricoxidesynthase ,NOS)活性变化 ;③阴茎组织中ET 1(endothelin 1)含量测定。结果 ①阴茎组织中NO含量先升高后降低 ,8月龄最高 ,2 4月龄最低 ,NOS活性变化与其一致 ,各月龄组间差别均非常显著 (P <0 0 1) ;cGMP含量表现为显著降低 (P <0 0 1) ;ET 1含量呈升高趋势 ,ET 1/NO比值也显著升高 (P <0 0 1) ;②L Arginine长时间喂养大鼠后 ,阴茎组织中NOS活性及NO、cGMP含量均显著增加 (P <0 0 1) ,ET 1含量无明显改变。结论 阴茎组织中cGMP含量及ET 1/NO比值可能决定着平滑肌细胞舒缩状态 ;L Arginine对增强NOS活性、增加NO、cGMP含量有明显作用 ,表明L Arginine有用于治疗勃功能障碍 (erectiledysfunction ,ED)

一氧化氮外源性供体L-arginine对破骨细胞骨吸收功能的影响

目的:一氧化氮在维持机体多个系统的生理功能中起重要作用,许多慢性疾病可造成一氧化氮产生减少,此时一氧化氮供体是一种必要的补充。观察一氧化氮外源性供体L-arginine对体外培养破骨细胞增殖及骨吸收功能的影响。方法:实验于2005-06/2006-05在四川大学华西医院生物治疗国家重点实验室干细胞与组织工程研究室完成。选择出生1d的清洁级SD大鼠乳鼠,采用骨髓诱导法体外培养破骨细胞,培养液内分别加入0.3,0.6,1.0g/L不同浓度的L-arginine,并以等体积三蒸水作为对照。培养7d后,以抗酒石酸酸性磷酸酶染色观察破骨细胞数目、形态,MIAS-2000图像分析仪检测骨片上骨吸收陷窝的数目和面积,并用扫描电镜观察不同浓度L-arginine对骨吸收陷窝的影响。结果:①破骨细胞的一般形态:破骨细胞较其他细胞大,形态不规则,呈油煎蛋形、长条形、腊肠形或漏斗形等,细胞内可见几个至几十个核不等。抗酒石酸酸性磷酸酶染色酶活性部分酒红色,颗粒状。②抗酒石酸酸性磷酸酶阳性细胞数:各组破骨细胞数目随着L-arginine浓度增加而减少(P<0.05)。③骨吸收陷窝的面积和数目:骨片培养7d,吸收陷窝计数的结果显示,0.3g/L以上浓度L-arginine对破骨细胞吸收功能均有明显抑制作用,并呈剂量相关性。0.3g/LL-arginine组陷窝面积为对照组的91%(P<0.05),0.6g/LL-arginine组为对照组的80%(P<0.05),1.0g/LL-arginine组为对照组的69%(P<0.01)。结论:采用骨髓诱导法培养的破骨细胞数量多、纯度高,且具有明显的骨吸收功能。L-arginine抑制破骨细胞增殖,抑制破骨细胞骨吸收功能,并呈剂量相关性。

BQ123与L-arginine在肝脏缺血再灌注损伤中的作用研究

目的 探讨肝脏缺血再灌注损伤的机制及 BQ12 3或 (和 ) L -arginine能否有效改善肝脏缺血再灌注损伤。方法 在大鼠肝脏缺血再灌注损伤模型基础上 ,对组织形态学、肝脏酶学、透明质酸、血浆内皮素及免疫组织化学染色情况进行观测。结果 肝脏缺血再灌注损伤时 ,血肝酶、透明质酶、血浆内皮素水平均显著增高 ,再灌注前使用 BQ12 3或 (和 ) L-arginine的肝脏 ,其组织结构及功能损伤均显著减轻。结论 肝脏缺血再灌注损伤与肝脏微循环改善有关 ,若能在肝脏再灌注前使用BQ12 3或 (和 ) L -arginine,可有效改善肝脏微循环 。

L-arginine对高糖诱导的内皮细胞衰老的作用

目的观察不同浓度的L-arginine对高糖诱导的人脐静脉内皮细胞（HUVECs）衰老的作用。方法HUVECs分别培养在正常糖浓度组（5．5mmol／L）、高糖组（33mmol／L）及高糖＋不同浓度L-arginine（0．4、0．8、1．6、3．2mmol／L）组，SAβ-gal活性评定细胞衰老的程度，PCR-ELISA法检测端粒酶活性，流式细胞术测定ROS及细胞周期，ELISA检测NO水平。结果与正常糖浓度组比较，高糖组SAβ-gal活性增强（P〈0．01），G0／G1期细胞比率增加（P〈0．01），端粒酶活性减弱（P〈0．01），细胞内ROS增多（P〈0．01），而NO水平减少（P〈0．001）。高糖环境下0．4—1．6mmol／L的L—arginine均可抑制SAβ—gal活性（P〈0．01），减少GD0/G0期细胞比率（P〈0．01），增强端粒酶活性（P〈0．01），减少细胞内ROS的生成（P〈0．01），增加NO水平（P〈0．01）。结论高糖可以诱导HUVECs的衰老。L-arginine可以延缓高糖诱导的内皮细胞衰老进程，但并没有明显的剂量依赖性。L—arginine可通过增强端粒酶活性，减少氧化应激，增加NO水平发挥抗内皮细胞衰老的作用。

The Potential Pathway of L-arginine·L-aspartate for Inhibition of Platelet Function

Aim L-Arginine· L-aspartate, a double salt, has been recently reported toinhibit platelet aggregation and thrombosis, but its action mechanism is not clear yet. This studywas conducted to investigate its effect on FITC-PAC-1, an anti-glycoprotein IIb/IIIa monoclonalantibody binding to activated platelets, and on correlative autacoid levels in plasma or inplatelets in order to explore its potential pathway of inhibiting platelet aggregation andthrombosis. Methods Monoclonal antibody binding to activated platelets was assayed by flowcytometry; NO was assessed by colorimetric method. cAMP, TXB_2 or 6-keto-PGF_(1α) levels wereassessed by radioimmunoassay. Results Gavaged 30 mg·kg^(-1) of L-arginine·L-aspartate increasedboth concentration of NO in plasma and 6-keto-PGF_(1) in incubated supernatant of aortic segment ofrats ex vivo (P < 0.05), but it did not influence cAMP content in platelets and the level of TXB_2or 6-keto-PGF_(1) in plasma of rats, whereas ASA significantly lowered TXB_2 or 6-keto-PGF_(1α) inplasma. Both 100 μmol-L^(-1) of L-arginine ·L-aspartate and ASA inhibited FITC-PAC-1 binding toactivated platelets in vitro. Conclusion The increase in NO and PGI_2 release from endo-thelialcells and consequent inhibition of platelet activation may contribute to the inhibition of plateletaggregation and thrombosis by L-arginine· L-aspartate; whereas arachidonic acid or cAMP metabolicpathway is not closely correlative with the studied effect.

L-arginine在高原鼠肝脏缺血再灌注损伤时肠道细菌易位中的作用研究

目的:探讨肝脏缺血再灌注损伤时肠道细菌易位的机制及精氨酸能否有效改善肠道细菌易位。方法:在大鼠肝脏缺血再灌注损伤模型基础上,对门静脉血、回肠系膜淋巴结进行肠道细菌培养。结果:肝脏缺血再灌注损伤时,血液及淋巴结可能培养出大肠埃希氏菌。但精氨酸预处理组与相同阻断时间组之间无差异。结论:高原地区第一肝门阻断的时间对肠道细菌易位有显著影响,但精氨酸对肝缺血再灌注损伤的肠道细菌易位无明显的保护作用。

L-Arginine对蛛网膜下腔出血大鼠脑组织水电解质含量的影响

目的探讨一氧化氮(nitricoxide,NO)合成底物L -arginine(L -Arg)对蛛网膜下腔出血 (subarachnoidhemorrhage,SAH)后继发性脑损害的保护作用。方法采用血管内穿刺法刺破脑底动脉环建立大鼠SAH模型 ,将动物随机分为SAH组和SAH +L -Arg组。L -Arg(Sigma)于术前30min按0.5g/kg腹腔注射 ,每隔6h以同样的剂量追加1次。在立体定向仪控制下 ,用激光多普勒血流计检测24h内大脑顶叶皮层局部脑血流量 (regionalcerebralbloodflow,rCBF) ;将动物在不同时间点处死后用干湿重比较法测定脑组织含水量 ,原子吸收分光光度计测量不同时间点脑组织Na +、K +和Ca2 +含量。股动脉插管监测血压和血气。结果SAH组术后rCBF迅速降低 ,1h达最低值 ,24h内无明显恢复 ;SAH后6h、24h脑组织含水率和Na +含量明显增加 ,K +含量减低 ;脑组织Ca2 +含量在SAH后1h开始显著增加,并持续24h。SAH +L -Arg组rCBF下降的速度减慢 ,下降的程度减轻 ;脑组织水、Na +含量增加的程度减轻 ,K +丢失减少 ,同时脑组织Ca2 +积聚减轻。结论NO合成底物L

Alterations of intestinal immune function and regulatory effects of L-arginine in experimental severe acute pancreatitis rats

AIM： To discuss the changes of intestinal mucosal immune function in rats with experimental severe acute pancreatitis （SAP） and the regulatory effect of L-arginine. METHODS： Male adult Wistar rats were randomly divided into pancreatitis group, sham-operation group, and L-arginine treatment group. Animals were killed at 24, 48, and 72 h after SAP models were developed and specimens were harvested. Endotoxin concentration in portal vein was determined by limulus endotoxin analysis kit. CD3＋, CD4＋, CD8＋ T lymphoo/tes in intestinal mucosal lamina propria were examined by immunohistochemistry. Secretory immunoglobulin A （SIgA） in cecum feces was examined by radioimmunoassay. RESULTS： Compared to the control group, plasma endotoxin concentration in the portal vein increased, percentage of CD3＋ and CD4＋ T lymphocyte subsets in the end of intestinal mucosal lamina propria reduced significantly, CD4＋/CD8＋ ratio decreased, and SIgA concentrations in cecum feces reduced at 24, 48, and 72 h after SAP developed. Compared to SAP group, the L-arginine treatment group had a lower level of plasma endotoxin concentration in the portal vein, a higher CD3＋ and CD4＋ T lymphocyte percentage in the end of intestinal mucosal lamina propria, an increased ratio of CD4＋/CD8＋ and a higher SIgA concentration in cecum feces. CONCLUSION： Intestinal immune suppression occurs in the early stage of SAP rats, which may be the main reason for bacterial and endotoxin translocation. L-arginine can improve the intestinal immunity and reduce bacterial and endotoxin translocation in SAP rats.

The Synthesis of Vasoactive Protected L-Arginine

Under the catalysis of dioxygenase L-arginine is converted to L-citrulline and nitric oxide,the latter exhibits endothelium derived relaxing factor(EDRF)-like actions.N^G-nitro-L-arginine has an inhibitory effect on the biosynthesis of EDRF in vitro and in vivo,hence it is an EDRF antagonist.The results of the present work indicate that both N^G-NO_2-L-Arg-OH and HCl·N^G-NO_2-L- Arg-OCH_3 have vasodilating effect in vitro,but produced dose-depending increase in mean arterial blood pressure(MAP)in vivo.In vitro HCl·N_G-NO_2-L-Arg-N_G-NO_2-L-Arg-OCH_3 relaxed rat aortic strip pretreated with noradrenaline(NE).In vivo,however,it produced biphasic effect,i.e,decreased MAP at lower dose and increases MAP at higher dose, N_G-Tos-L-Arg-N_G-Tos-L-Arg-OH produced dose-depending vasodilating and hypotensive actions.

L-arginine administration ameliorates serum and pulmonary cytokine response after gut ischemia-reperfusion in immature rats

AIM: Small intestinal ischemia-reperfusion (IR) has been demonstrated to result in both local mucosal injury and systemic injuries. The exact role of nitric oxide (NO) in intestinal IR is unclear. We propose that NO and some other cytokines change in the reperfusion period and these changes are associated with lung injury. The aim of this study was to determine the effect of supplementing NO substrate, L-arginine (L-arg), on serum and pulmonary cytokine production during small intestinal IR in immature rats. METHODS: Immature rats underwent 60 min. of superior mesenteric artery occlusion followed by 90 min of reperfusion. L-arg (250 mg/kg) was given intravenously to the experimental group (IR+L-arg) which received L-arg after 45 min of intestinal ischemia. Serum and lung endothelin-1 (ET-1), NO, malondialdehyde (MDA), and tumor necrosis factor a (TNFα) were measured. Sham operation (SHAM) and intestinal IR (IR) groups were performed as control. The lavage fluid of the lung was collected by bronchoalveolar lavage (BAL) and white blood cells and polymorphonuclear cells (PMNs) were immediately counted to identify lung damage. RESULTS: When L-arg was given during small intestinal IR, serum NO concentration increased significantly in IR+L-arg group (162.17±42.93 μmol/L) when compared with IR group (87.57±23.17 μmol/L, t=3.190, P= 0.008 <0.01). Serum MDA reduced significantly in IR+L-arg group (8.93±1.50 nmol/L) when compared with SHAM (23.78±7.81 nmol/L, t= 3.243, P= 0.007<0.01) and IR (25.54±9.32 nmol/L, t= 3.421, P= 0.006<0.01). ET-1 level in lung tissues was significantly lower in IR+L-arg group (13.81±7.84 pg/mL) than that in SHAM (35.52±10.82 pg/mL, t= 2,571, P= 0,03<0.05) and IR (50.83±22.05 pg/mL, t= 3.025, P= 0.009<0.01) groups. MDA contents in lung tissues were significantly lower in IR+L-arg group (10.73±1.99 nmol/L) than in SHAM (16.62±2.28 nmol/L, t= 3.280, P = 0.007<0.01) and IR (21.90±4.82 nmol/L, t= 3.322, P= 0.007<0.01) groups. Serum and lung TNFα concentrations were not significantly different in three groups. NO contents in lung homogenates and white blood cell counts in BAL had no significant difference in three groups; but the percentage of PMNs in BAL was 13.50±8.92, 33.20±16.59, and 22.50±6.09 in SHAM, IR, and IR+L-arg groups, respectively. CONCLUSION: Small intestinal IR induced increases of pulmonary neutrophil infiltration in immature rats. Neutrophil infiltration in lung tissues was reduced by L-arg administration but remained higher than in SHAM group. L-arg administration during intestinal IR enhances serum NO production, reduces serum MDA and lung ET-1 and MDA levels, resulting in the improvement of systemic endothelial function. L-arg supplementation before reperfusion may act as a useful clinical adjunct in the management of intestinal IR, thus preventing the development of adult respiratory distress syndrome, even multiple organ dysfunction syndrome (MODS).

Protective effects of L-arginine on reperfusion injury after pancreaticoduodenal transplantation in rats

BACKGROUND: Post-transplantation pancreatitis andgraft thrombosis are two major complications of pancreastrans-plantation that contribute to morbidity, mortality, andgraft loss. Nitric oxide (NO) is a potent vasodilator agentformed when L-arginine ( L-Arg) is converted to L-citrul-line by the action of NO synthase (NOS), and plays a ma-jor role in microcirculatory changes. We therefore investi-gated the effect of L-Arg on reperfusion injury followingpancreaticoduodenal transplantation in rats.METHODS: The homologous male Wistar rat model ofheterotopic total pancreaticoduodenal transplantation wasused. The L-Arg-treated rats received the intravenous in-jection of L-Arg 5 minutes before and after reperfusion at adose of 200 mg/kg while the N-Nitro-L-arginine methyl es-ter (L-NAME) -treated rats at a dose of 10 mg/kg. Theamount of NO in the pancreas graft was measured. Serumconcentration of cytokine-induced neutrophil chemoattrac-tant ( CINC) was determined by enzyme-linked immu-nosorbant assay, the expression of CINC mRNA was detect-ed by Northern blot assay in the pancreas graft, and the ac-tivity of myeloperoxidase (MPO) was measured. Histolo-gical examination was performed.RESULTS: The amount of NO was higher in the L-Arggroup than in the control group, while it was lower in theL-NAME group than in the control group (P <0.05). Thepeak of serum CINC concentration occurred 3 hours afterreperfusion with the difference among the groups being sig-nificant. The expression peak of CINC mRNA in the pan-creas graft occurred 3 hours after reperfusion. The expres-sion level in the L-Arg group (7.66 ± 1.53 μg/L) was lowerthan in the control group (26.31±2.01 μg/L), while in theL-NAME group (34.18 ±3.12 μg/L) it was higher than thatin the control group (P <0. 05). The activity of MPO inthe L-Arg group was obviously decreasd as compared within the other groups. The pancreas inflammation was ame-liorated when L-Arg was administered, whereas the panc-reas damage was aggravated when L-NAME was adminis-tered.CONCLUSIONS: L-Arg can increase the amount of NOand inhibit the elevation of CINC, the CINC mRNA ex-pression and early neutrophil accumulation in the pancreas.NO has protective effects on ischemia/reperfusion injury inpancreaticoduodenal transplantation.

Dynamic alterations in early intestinal development,microbiota and metabolome induced by in ovo feeding of L-arginine in a layer chick model

Background:Prenatal nutrition is crucial for embryonic development and neonatal growth,and has the potential to be a main determinant of life-long health.In the present study,we used a layer chick model to investigate the effects of in ovo feeding(IOF)of L-arginine(Arg)on growth,intestinal development,intestinal microbiota and metabolism.The treatments included the non-injected control,saline-injected control,and saline containing 2,6,or 10 mg Arg groups.Results:IOF Arg increased early intestinal index and villus height,and enhanced uptake of residual yolk lipid,contributing to subsequent improvement in the early growth performance of chicks.Prenatal Arg supplementation also increased the early microbialα-diversity,the relative abundance of Lactobacillales and Clostridiales,and decreased the relative abundance of Proteobacteria of cecum in chicks.Furthermore,the shift of cecal microbiota composition and the colonization of potential probiotics were accelerated by IOF of Arg.Simultaneously,metabolomics showed that metabolisms of galactose,taurine-conjugated bile acids and lipids were modulated to direct more energy and nutrients towards rapid growth of intestine at the beginning of post-hatch when embryos received IOF of Arg.Conclusions:Prenatal Arg supplementation showed beneficial effects on the early intestinal development,cecal microbiota and host metabolism of layer chicks,contributing to subsequent improvement in the early growth performance.These findings provide new insight into the role of IOF of Arg in the establishment of the gut microbiota of newly-hatched layer chicks,and can expand our fundamental knowledge about prenatal nutrition,early bacterial colonization and intestinal development in neonate.

Competitive metabolism of L-arginine:arginase as a therapeutic target in asthma

Exhaled breath nitric oxide （NO） is an accepted asthma biomarker. Lung concentrations of NO and its amino acid precursor, L-arginine, are regulated by the relative expressions of the NO synthase （NOS） and arginase isoforms. Increased expression of arginase I and NOS2 occurs in murine models of allergic asthma and in biopsies of asthmatic airways. Although clinical trials involving the inhibition of NO-producing enzymes have shown mixed results, small molecule arginase inhibitors have shown potential as a therapeutic intervention in animal and cell culture models. Their transition to clinical trials is hampered by concerns regarding their safety and potential tox- icity. In this review, we discuss the paradigm of arginase and NOS competition for their substrate L-arginine in the asthmatic airway. We address the functional role of L-arginine in inflammation and the potential role of arginase inhibitors as therapeutics.

Increased L-arginine Production by Site-directed Mutagenesis of N-acetyl-L-glutamate Kinase and pro B Gene Deletion in Corynebacterium crenatum

Objective In Corynebacterium crenatum,the adjacent D311 and D312 of N-acetyl-L-glutamate kinase(NAGK),as a key rate-limiting enzyme of L-arginine biosynthesis under substrate regulatory control by arginine,were initially replaced with two arginine residues to investigate the L-arginine feedback inhibition for NAGK.Methods NAGK enzyme expression was evaluated using a plasmid-based method.Homologous recombination was employed to eliminate the pro B.Results The IC50 and enzyme activity of NAGK M4,in which the D311 R and D312 R amino acid substitutions were combined with the previously reported E19 R and H26 E substitutions,were 3.7-fold and 14.6% higher,respectively,than those of the wild-type NAGK.NAGK M4 was successfully introduced into the C.crenatum MT genome without any genetic markers;the L-arginine yield of C.crenatum MT-M4 was 26.2% higher than that of C.crenatum MT.To further improve upon the L-arginine yield,we constructed the mutant C.crenatum MT-M4 ?pro B.The optimum concentration of L-proline was also investigated in order to determine its contribution to L-arginine yield.After L-proline was added to the medium at 10 mmol/L,the L-arginine yield reached 16.5 g/L after 108 h of shake-flask fermentation,approximately 70.1% higher than the yield attained using C.crenatum MT.Conclusion Feedback inhibition of L-arginine on NAGK in C.crenatum is clearly alleviated by the M4 mutation of NAGK,and deletion of the pro B in C.crenatum from MT to M4 results in a significant increase in arginine production.

Protective effects of L-arginine against ischemia-reperfusion injury in non-heart beating rat liver graft

BACKGROUND: Although the use of non-heart beating donors (NHBDs) could bridge the widening gap between organ demand and supply, its application to liver transplantation is limited due to the high incidence of primary graft loss. Prevention of liver injury in NHBDs will benefit the results of transplantation. This study was conducted to evaluate the protective effects of L-arginine on liver grafts from NHBDs. METHODS: One hundred and four Wistar rats were randomly divided into 7 groups: normal control (n=8) controls 1, 2 and 3 (C-1, C-2, C-3, n=16), and experimental 1, 2 and 3 (E-1, E-2, E-3, n=16). For groups C-1 and E-1, C-2 and E-2, and C-3 and E-3, the warm ischemia time was 0, 30, and 45 minutes, respectively. Liver grafts were flushed with and preserved in 4 degrees C Euro-collins solution containing 1 mmol/L L-arginine for 1 hour in each experimental group. Recipients of each experimental group were injected with L-arginine (10 mg/kg body weight) by tail vein 10 minutes before portal vein reperfusion. Donors and recipients of each experimental control group were treated with normal saline. Then transplantation was performed. At 1, 3, and 24 hours after portal vein reperfusion, blood samples were obtained to determine the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO) and plasma endothelin (ET). At 3 hours after portal vein reperfusion, grafts samples were fixed in 2.5% glutaraldehyde for electron microscopic observation. RESULTS: At I hour after portal vein reperfusion, the levels of NO in groups E-1, E-2, E-3 and C-1, C-2, C-3 were lower, while the levels of plasma ET, serum ALT and AST were higher than those in the normal control group (P<0.05). At 1, 3, and 24 hours, the levels of NO in groups E-1, E-2, E-3 were higher, while the levels of plasma ET, serum ALT and AST were lower than those in the corresponding control groups (C-1, C-2, C-3) (P<0.05). The levels of NO in groups C-2 and C-3 were lower than in group C-1 (P<0.05), and the level of NO in group C-3 was lower than in group C-2 (P<0.05). At 1, 3 and 24 hours, the levels of plasma ET, serum ALT, and AST in groups E-1, E-2, E-3 were lower than those in the corresponding control groups (C-1, C-2, C-3) (P<0.05). The levels of plasma ET, serum ALT, and AST were lower in group C-3 than in groups C-1 and C-2 (P<0.05). Pathological changes in groups E-1, E-2, E-3 were milder than those in the corresponding experimental control groups (C-1, C-2, C-3). CONCLUSIONS: The imbalance between NO and ET plays an important role in the development of ischemia-reperfusion injury of liver grafts from NHBDs. L-arginine can attenuate injury in liver grafts from NHBDs by improving the balance between NO and ET.

Dietary L-arginine supplementation reduces lipid accretion by regulating fatty acid metabolism in Nile tilapia(Oreochromis niloticus)

Background: Excessive white fat accumulation in humans and other animals is associated with the development of multiple metabolic diseases. It is unknown whether dietary L-arginine supplementation reduces lipid deposition in high fat diet-fed Nile tilapia(Oreochromis niloticus).Results: In the present study, we found that dietary supplementation with 1% or 2% arginine decreased the deposition and concentration of fats in the liver;the concentrations of triglycerides, low-density lipoprotein, total cholesterol, and high-density lipoprotein in the serum;and the diameter of adipocytes in intraperitoneal adipose tissue. Compared with the un-supplementation control group, the hepatic activities of alanine aminotransferase,aspartate aminotransferase, and lactate dehydrogenase, and hepatic concentration of malondialdehyde were reduced but these for catalase and superoxide dismutase were enhanced by dietary supplementation with 2% arginine. Arginine supplementation reduced the total amounts of monounsaturated fatty acids, while increasing the total amounts of n-3 and n-6 polyunsaturated fatty acids in the liver. These effects of arginine were associated with reductions in mRNA levels for genes related to lipogenesis(sterol regulatory element-binding protein-1, acetyl-CoA carboxylase α, stearoyl-CoA desaturase, and fatty acid synthase) but increases in mRNA levels for genes involved in fatty acid β-oxidation(carnitine palmitoyltransferase 1α and peroxisome proliferator-activated receptor α). In addition, hepatic mRNA levels for Δ4 fatty acyl desaturase 2 and elongase 5 of very long-chain fatty acids were enhanced by arginine supplementation.Conclusion: These results revealed that dietary L-arginine supplementation to tilapia reduced high fat diet-induced fat deposition and fatty acid composition in the liver by regulating the expression of genes for lipid metabolism.

Esophagogastric anastomosis in rats: Improved healing by BPC 157 and L-arginine, aggravated by L-NAME

AIM To cure typically life-threatening esophagogastric anastomosis in rats, lacking anastomosis healing and sphincter function rescue, in particular. METHODS Because we assume esophagogastric fistulas represent a particular NO-system disability, we attempt to identify the benefits of anti-ulcer stable gastric pentadecapeptide BPC 157, which was in trials for ulcerative colitis and currently for multiple sclerosis, in rats with esophagocutaneous fistulas. Previously, BPC 157 therapies have promoted the healing of intestinal anastomosis and fistulas, and esophagitis and gastric lesions, along with rescued sphincter function. Additionally, BPC 157 particularly interacts with the NOsystem. In the 4 d after esophagogastric anastomosis creation, rats received medication(/kg intraperitoneallyonce daily: BPC 157(10 μg, 10 ng), L-NAME(5 mg), or L-arginine(100 mg) alone and/or combined or BPC 157(10 μg, 10 ng) in drinking water). For rats underwent esophagogastric anastomosis, daily assessment included progressive stomach damage(sum of the longest diameters, mm), esophagitis(scored 0-5), weak anastomosis(m L H2 O before leak), low pressure in esophagus at anastomosis and in the pyloric sphincter(cm H2O), progressive weight loss(g) and mortality. Immediate effect assessed blood vessels disappearance(scored 0-5) at the stomach surface immediately after anastomosis creation. RESULTS BPC 157(all regimens) fully counteracted the perilous disease course from the very beginning(i.e., with the BPC 157 bath, blood vessels remained present at the gastric surface after anastomosis creation) and eliminated mortality. Additionally, BPC 157 treatment in combination with L-NAME nullified any effect of L-NAME that otherwise intensified the regular course. Consistently, with worsening(with L-NAME administration) and amelioration(with L-arginine), either L-arginine amelioration prevails(attenuated esophageal and gastric lesions) or they counteract each other(L-NAME + L-arginine); with the addition of BPC 157(L-NAME + L-arginine + BPC 157), there was a marked beneficial effect. BPC 157 treatment for esophagogastric anastomosis, along with NOS-blocker L-NAME and/or NOS substrate L-arginine, demonstrated an innate NO-system disability(as observed with L-arginine effectiveness). BPC 157 distinctively affected corresponding events: worsening(obtained with L-NAME administration that was counteracted); or amelioration(L-arginine + BPC 157-rats correspond to BPC 157-rats).CONCLUSION Innate NO-system disability for esophagogastric anastomoses, including L-NAME-worsening, suggests that these effects could be corrected by L-arginine and almost completely eliminated by BPC 157 therapy.

Celecoxib-induced gastrointestinal, liver and brain lesions in rats, counteraction by BPC 157 or L-arginine, aggravation by L-NAME

To counteract/reveal celecoxib-induced toxicity and NO system involvement. METHODSCelecoxib (1 g/kg b.w. ip) was combined with therapy with stable gastric pentadecapeptide BPC 157 (known to inhibit these lesions, 10 μg/kg, 10 ng/kg, or 1 ng/kg ip) and L-arginine (100 mg/kg ip), as well as NOS blockade [N(G)-nitro-L-arginine methyl ester (L-NAME)] (5 mg/kg ip) given alone and/or combined immediately after celecoxib. Gastrointestinal, liver, and brain lesions and liver enzyme serum values in rats were assessed at 24 h and 48 h thereafter. RESULTSThis high-dose celecoxib administration, as a result of NO system dysfunction, led to gastric, liver, and brain lesions and increased liver enzyme serum values. The L-NAME-induced aggravation of the lesions was notable for gastric lesions, while in liver and brain lesions the beneficial effect of L-arginine was blunted. L-arginine counteracted gastric, liver and brain lesions. These findings support the NO system mechanism(s), both NO system agonization (L-arginine) and NO system antagonization (L-NAME), that on the whole are behind all of these COX phenomena. An even more complete antagonization was identified with BPC 157 (at both 24 h and 48 h). A beneficial effect was evident on all the increasingly negative effects of celecoxib and L-NAME application and in all the BPC 157 groups (L-arginine + BPC 157; L-NAME + BPC 157; L-NAME + L-arginine + BPC 157). Thus, these findings demonstrated that BPC 157 may equally counteract both COX-2 inhibition (counteracting the noxious effects of celecoxib on all lesions) and additional NOS blockade (equally counteracting the noxious effects of celecoxib + L-NAME). CONCLUSIONBPC 157 and L-arginine alleviate gastrointestinal, liver and brain lesions, redressing NSAIDs’ post-surgery application and NO system involvement.