Recent advances in diverse nanosystems for nitric oxide delivery in cancer therapy

Gas therapy has been proven to be a promising and advantageous treatment option for cancers. Studies have shown that nitric oxide (NO) is one of the smallest structurally significant gas molecules with great potential to suppress cancer. However, there is controversy and concern about its use as it exhibits the opposite physiological effects based on its levels in the tumor. Therefore, the anti-cancer mechanism of NO is the key to cancer treatment, and rationally designed NO delivery systems are crucial to the success of NO biomedical applications. This review summarizes the endogenous production of NO, its physiological mechanisms of action, the application of NO in cancer treatment, and nano-delivery systems for delivering NO donors. Moreover, it briefly reviews challenges in delivering NO from different nanoparticles and the issues associated with its combination treatment strategies. The advantages and challenges of various NO delivery platforms are recapitulated for possible transformation into clinical applications.

Nitric oxide-releasing drug delivery systems for overcoming drug resistance in chemotherapy / 药学学报

Chemotherapeutic agents along with other treatments, such as chemotherapy and radiotherapy, have made significant contributions to cancer therapy, however multidrug resistance (MDR) in tumor remains an important developmental barrier to efficient chemotherapy. In recent research, there is increasing evidence that nitric oxide (NO) has the potential to overcome MDR. Unlike other chemosensitizers that ameliorate MDR but are potentially toxic, NO is endogenous and biocompatible molecule, which makes it even more promising as a cancer therapeutic. Nanoparticle-based drug delivery systems not only facilitate the delivery of multiple therapeutic agents, but also promote the avoidance of MDR, which are promising to both efficient delivery of NO and anti-cancer drugs in combination. Therefore, this review will discuss the mechanisms how NO reverse MDR and the recent advances in the application of NO functionalized nanoparticles for anticancer drug delivery.

Regulation of nitric oxide donor JS-K on tumor energy metabolism in H22 tumor-bearing mice / 中国药理学与毒理学杂志

OBJECTIVE To investigate the regulation of{O2 (2,4-dinitrophenyl)1-[(4-ethoxycarbonyl) piperazin-1-yl] diazen-1-ium-1, 2-diolate} (JS-K), a nitric oxide donor, on tumor energy metabolism in H22 tumor-bearing mice. METHODS The hepatoma animal model in BALB/c mice was established with H22 cell line. The inoculated mice were randomly divided into four groups. The JS-K group and model group received JS-K (0.75 and 1.50 mg?kg-1) and saline via tail the vein intravenously once every 3 d for 14 d, and 5 injections, respectively. The fluorouracil (5-FU) group received 5-FU 20 mg·kg-1 by intra-peritoneal injection once a day for 14 d. On the 15th day after the first administration, mice were sacri-ficed and the tumor, thymus and spleen were isolated and weighed immediately. The tumor growth inhibitory rate and organ index were calculated. The activities of hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), succinate dehydrogenase (SDH), adenosinetriphosphatase (ATPase), and the levels of lactic acid (LD) and adenosine triphosphate (ATP) in tumor tissues were determined by colorimetric method. The expression of hypoxia-inducible factor 1 alpha (HIF-1α) and hexokinaseⅡ(HKⅡ) in the tumor tissue was analyzed by Western blotting. RESULTS Compared with model group, the tumor mass of JS-K 0.75 and 1.50 mg · kg-1 groups was significantly reduced (P<0.01), and the tumor growth inhibitory rate was 23.9%and 50.3%, respectively. There was no diffrence in thymic and splenic indexes between JS-K group and model group. The activity of HK, PFK, SDH, PK and ATPase of tumor tissue in model group was 22.6±3.7, 14.4±2.6, (10.5±2.6) U·g-1protein, (12.9±3.2) kU·g-1 protein and (0.70 ± 0.10) mmolPi · g-1protein · h-1, respectively, which dropped by 42.0%, 26.6%, 22.7%, 23.3%and 21.7%respectively (P<0.01, P<0.05) in JS-K 1.50 mg?kg-1 group. Compared with the model group, the level of ATP of tumor tissue in JS-K 1.50 mg?kg-1 groups dropped by 16.6%(P<0.01) and the level of LD in JS-K 0.75 and 1.50 mg?kg-1 groups dropped by 38.7%and 59.4%(P<0.01), respectively. In addi-tion, the expression of HIF-1αof tumor tissue in JS-K 1.50 mg?kg-1 group was decreased (P<0.01), and the expression of HKⅡ of tumor tissue in JS-K 0.75 and 1.50 mg?kg-1 groups was decreased signifi-cantly (P<0.05, P<0.01). CONCLUSION JS-K can inhibit the growth of tumor in H22 tumor-bearing mice and its mechanism may be related to regulating the tumor energy metabolism by inhibiting glycolysis and aerobic oxidation.

Nitric oxide donor PABA/NO induced mitochondrial-mediated apoptosis in HepG2 cells / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effects of nitric oxide (NO) donor, O2-{2,4-dinitro-5-[4-(N-methylamino) benzoyloxy]phenyl}1-(N,N-dimethylamino)diazen-1-ium-1,2-diolate (PABA/NO) on apop-tosis in human hepatocarcinoma cells. METHODS Proliferation of HepG2 cells treated with PABA/NO 7.5, 15.0 and 30.0μmol · L-1 was measured with Cell Counting Kit-8 (CCK-8) assay, the morphological features were observed under fluorescence microscopy, the level of NO was measured by DAF-FM DA staining, the apoptosis rate was determined by Annexin Ⅴ-FITC staining, mitochondrial membrane potential was determined by Rhodamine 123 staining, and protein expressions of Bcl-2, Bax, cleaved caspase 3, cytochrome c (Cyt c) and apoptosis inducing factor (AIF) were measured by Western blotting analysis. RESULTS Compared with cell control group, PABA/NO could obviously inhibit the proliferation of HepG2 cells (P<0.01). IC50 value of HepG2 cells treated with PABA/NO for 24 h was (10.8±0.6)μmol·L-1. The cells became round, deformed and appeared shrunken.The level of NO was increased and the fluores-cence intensity was 121 ± 9 (P<0.05), 174 ± 31 (P<0.05) and 230 ± 43 (P<0.01). The apoptosis rate was increased from (2.9 ± 0.5)％ to (17.0 ± 4.5)％ (P<0.01), (39.8 ± 5.4)％ (P<0.01) and (74.3 ± 45.2)％ (P<0.01). The fluorescence intensity of Rh123 was reduced from 668±69 to 605±73, 420±65 (P<0.05) and 242±47 (P<0.01). Compared with cell control group, PABA/NO down-regulated Bcl-2, up-regulated Bax and activated cleaved caspase 3. Meanwhile, the expression of Cyt c in the cytoplasm was increased from 0.15±0.04 to 0.27±0.06 (P<0.05), 0.38±0.07 (P<0.01) and 0.82±0.16 (P<0.01). The expression of AIF in the nucleus was increased from 0.183±0.032 to 0.231±0.011, 0.682±0.020 (P<0.01) and 0.966± 0.090 (P<0.01). Addition of carboxy-PTIO (NO scavenger) 50 μmol · L- 1 blocked PABA/NO-induced down-regulation of Bcl-2, up-regulation of Bax and cleaved caspase 3 activation. Additionally, up-regu-lation of Cyt c in the cytoplasm and up-regulation of AIF in the nucleus were also blocked by carboxy-PTIO in PABA/NO-treated HepG2 cells (P<0.01). CONCLUSION PABA/NO may induce HepG2 cell apoptosis through a mitochondrial pathway.

Regulation of nitric oxide donor JS-K on tumor energy metabolism in H22 tumor-bearing mice / 中国药理学与毒理学杂志

OBJECTIVE To investigate the regulation of {O2 (2,4-dinitrophenyl)1-〔(4-ethoxycarbonyl) piperazin-1-yl〕diazen-1-ium-1,2-diolate}(JS-K), anitric oxide donor, on tumor energy metabolism in H22 tumor- bearing mice. METHODS The hepatoma animal model in BALB/c mice was established with H22 cell line. The JS-K group and model group were received JS-K (0.75 and 1.5 mg?kg-1) and saline via tail intravenous once every 3 d for 14 d, received 5 injections, respectively. The positive group was received 5-FU 20 mg·kg- 1 by intraperitoneal injection once a day for 14 d. On the 15th day mice were sacrificed. The tumor growth inhibition rate were calculated. The activities of hexokinase (HK), phospho?fructo kinase (PFK), pyruvate kinase (PK), succinate dehydrogenase (SDH), adenosine triphosphatase (ATPase), and the levels of lactic acid (LD) and adenosine triphosphate (ATP) in tumor tissues were de?termined by colorimetric method. RESULTS Compared with model group, the tumor mass of JS- K 0.75 and 1.5 mg·kg- 1group was significantly reduced (P<0.01),and the tumor growth inhibition rate was 23.9% and 50.3%, respectively. The activity of HK, PFK, PK, SDH and ATPase of tumor tissue in model group was (22.6±3.7, 14.4±2.6, 12.9±3.2 and 10.5±2.6)U·g-1 protein and (0.70±0.10)μmolPi·mg-1 protein per hour, respectively; which in JS-K 1.5 mg?kg-1 group was dropped by 42.0%, 26.6%, 22.7%, 23.3% and 21.7% (P<0.01, P<0.05). Compared with the model group, the level of ATP and LD in JS-K group was dropped (P<0.01). CONCLUSION JS-K can inhibit the growth of tumor in H22 tumor-bearing mice and its mechanism may be related to regulating the tumor energy metabolism with inhibition of glycolysis and aerobic oxidation.

New nitric oxide donors based on ruthenium complexes

Nitric oxide (NO) donors produce NO-related activity when applied to biological systems. Among its diverse functions, NO has been implicated in vascular smooth muscle relaxation. Despite the great importance of NO in biological systems, its pharmacological and physiological studies have been limited due to its high reactivity and short half-life. In this review we will focus on our recent investigations of nitrosyl ruthenium complexes as NO-delivery agents and their effects on vascular smooth muscle cell relaxation. The high affinity of ruthenium for NO is a marked feature of its chemistry. The main signaling pathway responsible for the vascular relaxation induced by NO involves the activation of soluble guanylyl-cyclase, with subsequent accumulation of cGMP and activation of cGMP-dependent protein kinase. This in turn can activate several proteins such as K+ channels as well as induce vasodilatation by a decrease in cytosolic Ca2+. Oxidative stress and associated oxidative damage are mediators of vascular damage in several cardiovascular diseases, including hypertension. The increased production of the superoxide anion (O2-) by the vascular wall has been observed in different animal models of hypertension. Vascular relaxation to the endogenous NO-related response or to NO released from NO deliverers is impaired in vessels from renal hypertensive (2K-1C) rats. A growing amount of evidence supports the possibility that increased NO inactivation by excess O2- may account for the decreased NO bioavailability and vascular dysfunction in hypertension.

Therapeutic effects of ZK_(14), a novel nitric oxide donating biphenyldicarboxylate derivative, on hepatic fibrosis in rats / 中国药科大学学报

Aim: To study the therapeutic effects of ZK_(14), a novel nitric oxide donor, on hepatic fibrosis in rats. Methods: Rats were injected intraperitoneally with CCl_4 and induced to hepatic fibrosis, followed by intra-gasrtic administration with high dose of 20 mg/kg and low dose of 10 mg/kg of ZK_(14) for 4 weeks. The effects were evaluated in the content of nitric oxide of serum and nitric oxide synthases of liver, in serum levels of the hepatic functions indices and hepatic fibrotic index and in liver histopathology. Results: The biochemical indices and pathology grade were effectively improved in the group of ZK_(14) 20 mg/kg, and liver function was also effectively protected in this group. Conclusion: ZK_(14), a novel nitric oxide donating biphenyldicarboxylate derivative, showed preferable therapeutic effects on hepatic fibrosis of rats induced with CC14, and it may be of potential value in the treatment of hepatic fibrosis.

Preventive Effects of Nitroglycerine on Glucocorticoid-induced Osteoporosis in Growing Rats / 华中科技大学学报（医学）（英德文版）

The preventive effects of nitroglycerine (NG) on glucocorticoid-induced osteoporosis in growing rats were studied. Three-month-old female Wistar rats were randomly divided into control group (CON), dexamethasone group (DXM), DXM plus a low dose NG group (NG-L), DXM plus a middle dose NG group (NG-M) and DXM plus a high dose NG group (NG-H), 8 rats in each group. The rat model of osteoporosis was developed by intramuscular injection of dexamethasone twice a week. NG 0.2, 0.4 and 1.0 mg/kg was administered by oral gavages to the treatment groups every day for 12 weeks. Rats in CON group and DXM group were treated with normal saline of the same amount. After the treatment, the bone mineral density (BMD) and bone metabolism-associated bio-chemical markers were determined. Compared with CON group, BMD of lumbar spine and femur in DXM group was decreased significantly (P<0.05 and P<0.01 respectively), blood BGP levels and NO levels reduced (both P<0.01), and TRAP level increased (P<0.05). As compared with DXM group, BMD, serum BGP and NO were increased, and TRAP decreased in NG-L group and NG-M group, but had no significant difference in comparison to CON group. All the markers other than se- rum NO and TRAP levels had no significant difference between NG-H group and DXM group.It was concluded that low or middle doses of NG could prevent glucocorticoid-induced bone loss in growing rats, but high dose of NG could not. Supplement with NO donor could be considered as a preventive treatment for glucocorticoid-induced osteoporosis in a developing skeleton.

Effects of nebulized nitric oxide donors on acute hypoxic pulmonary hypertension in newborn piglets / 中华急诊医学杂志

Objective To observe the effects of nebulized nitric oxide donors on pulmonary and systemic hemodynamics during pulmonary hypertension induced by hypoxia (FiO_2=0.1,for 1 hour)in newborn piglets. Methods Twenty-four anesthetized and mechanically ventilated piglets were randomly divided into four groups:(1)group S (n=6),control group;(2)group C (n=6),receiving saline after hypoxia;(3)group NTG,receiving nebulized nitroglycerin after hypoxia;(4)group SNP,receiving nebulized sodium nitroprusside after hypoxia. Mean arterial pressure (MAP)and mean pulmonary arterial pressure (MPAP)were monitored continuously. Arterial blood gas analysis was determined at baseline,1 hour after hypoxia,termination of nebulization and 0.5 h after nebulization. After nebulization,blood methemoglobin (MetHb)and plasma NO level were measured.Results Acute hypoxia resulted in a significant increase in MPAP. At 1 hour after hypoxia,MPAP in group C,group NTG and group SNP were significantly higher than that in group S(P0.05).After nebulization,there were no significant differences in plasma NO level and blood MetHb level between groups. No rebound pulmonary hypertension was observed after withdrawal of nebulization. Conclusion Both NTG and SNP nebulization could be used as selective pulmonary vasodilators for acute hypoxic pulmonary hypertension.

Regulation of nitric oxide on glucocorticoid receptor function in rat alveolar macrophages treated by LPS in vitro / 第三军医大学学报

Objective To investigate the effects of nitric oxide donor (SNP) on glucocorticoid receptor function in condition of cell inflammation. Methods After fluorescence expression plasmid pGFP-GR transfected rat alveolar macrophges (AMs), nuclear translocation of GFP-GR following to lipopolysaccharide (LPS) and SNP treatment was observed through fluorescence microscope. The transcriptional activation activity of GR was evaluated by the method of relative activity of luciferase. Electrophoretic mobility shift assays (EMSA) were used to measure the activity of NF-?B. Results GFP-GR showed nuclear translocation in 2 h after nitric oxide donor (500 ?mol/L SNP) treatment, and the transcriptional activation activity of GR increased and the activity of NF-?B decreased remarkedly. The phenomena of depressed activity of NF-?B disappeared after the addition of GR special antagonism RU486. Conclusion Nitric oxide donor (500 ?mol/L SNP) exerts anti-inflammatory effect through GR activation in condition of cell inflammation.

Research progress and clinical perspectives of the nitric oxide donor drugs / 中国临床药理学与治疗学

The nitric oxide donor drugs have the functions of anti-hyperplasia, inhibiting platelet adhesion and aggregation, regulating immune response, etc. It suggests that these drugs play important roles in therapy of cardiovascular diseases, male impotence and pregnancy-induced hypertension syndrome.

Effects of sodium nitroprusside (SNP) on TLR4 in U937 induced by LPS / 中国免疫学杂志

Objective:To investigate the effects of nitric oxide donor,sodium nitroprusside(SNP)on the expression and production of TLR4 in U937 induced by LPS.Methods:U937 was induced to maturation by PMA and then stimulated by LPS.Then the cells were treated with SNP and divided into four groups:control group without stimulation of LPS,LPS group(10 ng/ml LPS+100 ng/ml rhLBP),low dose SNP group and high dose SNP group(50,500 ?mol/L SNP respectly).The mRNA and protein of TLR4 was determined by RT-PCR and Western blot.Results:The mRNA of TLR4 in SNP groups was lower than those in LPS group(0.308?0.050 and 0.138?0.0044 vs 0.342?0.098,P0.05).Conclusion:SNP might have a potential protective role in LPS induced inflammation such as sepsis and acute lung injury through inhibiting the mRNA and protein expression of TLR4.

The Effects of Local Nitric Oxide Donor Delivery in Stented Patients

BACKGROUND: The endovascular stent has been applied clinically in acute arterial occlusions after intimal dissection by angioplasty and in the prevention of restenosis. However, subacute stent thrombosis and restnosis remain major concerns in clinical stenting despite intravscular ultrasound guidance and high pressure inflation. Moreover, anticoagulation before and after stent implantation may be required for long periods and complicated by bleeding. A new strategy may be local drug delivery, which maintains sustained local concentration and may limit systemic complications. To evaluate the efficacy of local Nitric Oxide(NO) donor delivery on acute or subacute stent thrombosis and bleeding complications in patients, local NO donor delivery was performed in stented patients. METHOD: NO donor (2.0mg, Molsidomine) was delivered (1.0ml/min over 10min) using the Dispatch Catheter, after predilation of target lesion in 15 patients (8 angina, 7 myocardial infarction, mean age 5311.5 yr.) without heparin or nitrate infusion after stenting. After local NO donor delivery, Palmaz-Schatz stents were placed with standard methods. APTT and CK were checked at 1 hr, 3 hrs and 24 hrs after local NO donor delivery and STENTING. Follow-up coronary angiograms were done 48 hrs after stenting. RESULT: All patients had no hypotensive effects, no ischemic symptoms or no ECG changes during and after locaL NO donor delivery. ARTT and CK values were not changed at 3 and 24 hrs after local NO donor delivery and stenting. This allowed early arterial sheath removal. Follow-up coronary angiograms at 48 hrs showed all stents patent without stent recoil, with TIMI III flow, and without intra-stent thrombus. No target lesion revascularization and 100% event free survival were obsered for one month's clinical follow-up after NO donor delivery and stenting. Conclusion: Local NO donor delivery prior to stenting prevents acute and subacute stent thrombosis, systmic complications of nitrate, and maintains stent blood flow without stent recoil within the first one month after stenting.