OBJECTIVETo investigate the effects of aminoguanidine cream on the proliferation of keratinocytes (KC), content of advanced glycosylation end products (AGE) and oxidative stress in skin tissue of rats with diabetes.

METHODSStearic acid, liquid paraffin, vaseline, lanolin, isopropyl myristate fat, glycerol, 50 g/L alcohol paraben, aminoguanidine hydrochloride etc. were mixed in certain proportion to make aminoguanidine cream, and cream without aminoguanidine was used as matrix. The dorsal skin of normal rats were harvested and treated by aminoguanidine cream with dose of 5, 10 g/L, or 5 g/L together with 10 g/L azone. The transdermal effect was respectively measured at post treatment hour 2, 4, 7, 10, 12, 24. Thirty SD rats were divided into normal control (NC, n = 6), diabetes (D, n = 8), aminoguanidine cream-interfered (AI, n = 8), matrix cream-interfered groups (MI, n = 8) according to the random number table. Diabetes was reproduced by intraperitoneal injection of STZ (65 mg/kg) in rats of D, AI, and MI groups, and rats in NC group were injected with 0.05 mmol/L citrate buffer as control. One week later, dorsal skin of rats in AI and MI groups were respectively treated with 10 g/L aminoguanidine cream and matrix cream by external use for 4 weeks. AGE content was determined with fluorescence detection from skin collagen extract. KC cell cycle was detected by flow cytometry. Skin tissue specimens were obtained for determination of levels of superoxide dismutase (SOD), malondialdehyde (MDA), myeloperoxidase (MPO), and total antioxidant capacity. Data were processed with t test.

RESULTSTransdermal effect of aminoguanidine cream with dose of 10 g/L was better than that with 5 g/L or 5 g/L + 10 g/L azone cream. One rat was not induced successfully in MI group. Four weeks after model reproduction, 4 rats died in D group and 1 rat died in AI group. The AGE content in D group was obviously higher than that in NC group [(36.8 +/- 2.6), (24.6 +/- 2.7) U per milligram hydroxyproline, respectively, t = 7.2, P < 0.01], and that in AI group [(28.6 +/- 3.7) U per milligram hydroxyproline] was also lower as compared with that in D group (t = -3.9, P < 0.05). There was no significant difference in AGE content between MI [(32.2 +/- 5.2) U per milligram hydroxyproline] and D groups (t = 1.6, P > 0.05). The percentage of KC in S phase was obviously lower in D group than in NC group [(5.3 +/- 0.6)%, (7.6 +/- 0.9)%, respectively, t = 4.50, P < 0.01], while that in MI group [(9.2 +/- 1.5)%] was higher as compared with that in D group ( t = 4.90, P < 0.01). It was more higher in AI group than in D group on KC percentage in S and G2/M phase (with t value respectively 6.80, 3.17, P values all below 0.01). The oxidative stress indexes of skin tissue in D group were all higher than those in NC group, in which levels of MPO and SOD showed statistical difference (with t value respectively 4.4, 3.7, P values all below 0.05). The oxidative stress indexes were all lower in AI group than in D group, especially in SOD level (t = -1.4, P < 0.05). Levels of MAD, MPO in MI group were significantly lower than those in D group (with t value respectively 2.6, 2.9, P values all below 0.05).

CONCLUSIONSAminoguanidine cream can promote KC proliferation and appropriately reduce oxidative stress through inhibiting AGE formation to a certain extent in skin tissue of rats with diabetes. Signal use of matrix cream can also reduce oxidative stress in skin tissue of rats with diabetes.

Administration, Cutaneous ; Animals ; Cell Proliferation ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Glycation End Products, Advanced ; metabolism ; Guanidines ; administration & dosage ; pharmacology ; Keratinocytes ; drug effects ; Male ; Ointments ; administration & dosage ; pharmacology ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; metabolism ; pathology

AIMTo explore the effects of aminoguanidine (AG) inhalation on bleomycin (BLM)-induced fibrosis in lungs of rats and its possible mechanism.

METHODSSixty male Sprague-Dawley rats were randomly divided into 4 groups: BLM plus normal saline (NS) group, BLM plus 10 mmol/L AG group, BLM plus 50 mmol/L AG group, and NS plus NS group. At the same day when administrated by single intratracheal instillation of BLM (5 mg/kg) or equal volume of NS as control, the rats received NS (the same volume as AG) or AG inhalation (10 mmol/L AG, or 50 mmol/L AG, 5 min/each time, 2/day) for 30 d. The nitrite/nitrate (NO2-/NO3-) content of plasma in pulmonary artery, hydroxyproline content and the pathological changes in lungs, as well as lipid peroxide (LPO) content of plasma in pulmonary artery were examined.

RESULTSThe NO2-/NO3- content of plasma in pulmonary artery was increased in rats on day 14 after intratracheal instillation of BLM, compared with that of the control rats (P < 0.01). The hydroxyproline content in lung, the grade of pulmonary alveolitis and the content of LPO of plasma in pulmonary artery were increased in rats on day 30 after intratracheal instillation of BLM, compared with that of the control rats, respectively (P < 0.05, P < 0.01, P < 0.01). The above-mentioned changes were ameliorated by AG inhalation (10 mmol/L AG, or 50 mmol/ LAG, 5 min/each time, 2/day) for 30 d (P < 0.01, P < 0.05, P < 0.05, P < 0.01).

CONCLUSIONAG inhalation has anti-action on BLM-induced fibrosis in lung, which might be related to blockage of oxidative injury in lung.

Administration, Inhalation ; Animals ; Bleomycin ; Enzyme Inhibitors ; administration & dosage ; pharmacology ; Guanidines ; administration & dosage ; pharmacology ; Male ; Nitric Oxide Synthase ; antagonists & inhibitors ; Oxidative Stress ; drug effects ; Pulmonary Fibrosis ; chemically induced ; physiopathology ; prevention & control ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the role of advanced glycation end products (AGEs) and their receptors (RAGE) in the pathogenesis of diabetic mellitus erectile dysfunction (DMED) and the effects of AGEs and RAGE on the activity of endothelin-1 (ET-1) in rat cavernosum.

METHODSForty male Sprague-Dawley rats were taken at random to construct 2 groups of diabetes mellitus (DM) models of equal number, one given free access to water and the other administered aminoguanidine hydrochloride (DM + AG) in water at the dose of 1 g/L. Another 20 male SD rats were equally divided into a normal control and an AG control group. After 8 weeks, the cavernosum tissues were harvested from all groups of rats, part of the isolated penile tissues homogenated to detect the content of AGE-peptide (AGE-P) and the activity of ET-1, and the AGEs and RAGE in the rest of the penile tissues analyzed by immunohisto- chemical assay.

RESULTSCompared with the normal controls, the expressions of AGEs and RAGE, the content of AGE-P and the activity of ET-1 in the cavernosum tissues were significantly high in the DM group (P < 0.05), while the administration of AG to the DM rats reversed the above results. No significant difference was observed between the normal control and AG control groups in any of the data (P > 0.05).

CONCLUSIONIn DM conditions, the joint effect of AGEs and RAGE may elevate the activity of ET-1 in rat cavernosum and thus promote the development of DMED.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; physiopathology ; prevention & control ; Endothelin-1 ; metabolism ; Enzyme Inhibitors ; administration & dosage ; Glycation End Products, Advanced ; antagonists & inhibitors ; metabolism ; Guanidines ; administration & dosage ; Male ; Penis ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; metabolism

AIMTo investigate the effects of L-arginine and nitric oxide synthase (NOS) inhibitor Aminoguanidine (AG) on endotoxin induced lung injury in rats.

METHODSForty eight healthy male SD rats weighing (300 +/- 20) g were used. The animals were anesthetized with 20% urethane 1 g x kg(-1). Common carotid artery (CAA) and common carotid vein (CAV) were exposed through a median incision in the neck. Mean arterial pressure (MAP) was measured through a pressure transducer connected with intubation of CAA. The animals were randomly divided into six groups: group 1: control: group 2: LPS (5 mg x kg(-1) intravenous injection, i.v.); group 3: AG (50 mg x kg(-1) intraperitoneal injection, IP); group 4: high dose L-arginine (500 mg x kg(-1), IP); group 5: low dose L-arginine (250 mg x kg(-1) IP). Group 6: L-arginine + AG (250 mg x kg(-1), 50 mg x kg(-1), IP). Group 1: The animals were killed 6 h after 0.9% saline solution was given. Group 2: 0.9% saline solution was given 3 h after LPS i.v. and the animals were killed 3 h after medication. Group 3, 4, 5 and 6: AG, L-arginine and L-arginine+ AG were given 3 h after LPS i.v. respectively and the animals were killed 3 h after medication respectively. The pulmonary was removed immediately. The pulmonary coefficient and water content in pulmonary tissue were calculated (%). The NO content in plasma, MDA content and NOS, SOD activity in the pulmonary tissue were measured.

RESULTSL-arginine, AG and L-arginine + AG significantly decreased pulmonary coefficient and water content in pulmonary tissue and ameliorated endotoxin induced lung injury. AG and L-arginine + AG significantly decreased NO content in plasma, decreased MDA content and inhibited NOS activity and enhanced SOD activity in the pulmonary tissue.

CONCLUSIONIt may be concluded that L-arginine, AG and L-arginine + AG have beneficial effects on lung injury induced by LPS.

Animals ; Arginine ; administration & dosage ; therapeutic use ; Endotoxins ; adverse effects ; Guanidines ; administration & dosage ; therapeutic use ; Lung Injury ; chemically induced ; drug therapy ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVESTo explore the effects of aminoguanidine (AG) in different dosages on renal function in endotoxin induced rabbits shock model in the early stage and to approve the effects of dose-depended and time-depended of AG.

METHODSFourty New Zealand rabbits under anaesthesia were randomly divided into 5 groups: sham group, LPS group, the first group of AG, the second group of AG, the third group of AG. Each rabbits from the four groups received Escherichia Coli O55B5 LPS 400 micro g/kg to induce endotoxic shock except sham group. Edotoxic shock was diagnosed when the mean arterial pressure (MAP) decreased to 30%. Each rabbits in sham group and LPS group received 5ml NS, in the other three groups were infused with AG 30 mg/kg (the first group of AG), 50 mg/kg (the second group of AG), 100 mg/kg (the third group of AG) in 5ml NS, respectively. Urine output was recorded at the following time points, before injecting IPS (T(0)), shock (T), 1 h (T(1)), 2 h (T(2)), 3 h (T(3)), 4 h (T(4)), 5 h (T(5)) and 6 h (T(6)) after shock. Plasma nitrate and nitrite (NO(3)(-)/NO(2)(-), stable products of NO), BUN, Scr, RBP were determined at the time points of T, T(2), T(4) and T(6).

RESULTSLPS increased NO(3)(-)/NO(2)(-), BUN, Scr, RBP [from (47 +/- 5) micro mol/L, (5.8 +/- 1.5) mmol/L, (41 +/- 10) micro mol/L, (240 +/- 61) ng/L (T(0)) to (160 +/- 18) micro mol/L, (15.5 +/- 1.8) mmol/L, (166 +/- 23) micro mol/L, (1580 +/- 180) ng/L (T(6)), respectively, P < 0.01]; Urine output decreased significantly [from (17.6 +/- 2.8) ml (T(0)) to (1.3 +/- 0.6) ml (T(6)), P < 0.01]. AG attenuates the increasing of NO(3)(-)/NO(2)(-), BUN, Scr and RBP, and decreasing of urine output. NO(3)(-)/NO(2)(-) of the first, second and third group of AG at T(6) were (58 +/- 8), (50 +/- 14) and (46 +/- 9) micro mol/L, respectively. Compared to LPS group, there was a significant difference (P < 0.01). BUN was (8.2 +/- 2.9), (7.5 +/- 1.9) and (5.5 +/- 1.8) mmol/L, respectively at T(6). Compared to LPS group, there was a significant deference (P < 0.01). RBP was (350 +/- 60), (272 +/- 72) and (248 +/- 103) ng/L, respectively at T(6) (compared to LPS group, there was a significant deference. P < 0.05, < 0.05, < 0.01). Urine output was (11. 1 +/- 2.4), (12. 1 +/- 1. 3) and (17.1 +/- 2. 4) ml, respectively on T(6) (compared to LPS group, there was a significant deference, P < 0.01). AG of 100 mg/kg showed the best effect among three AG groups.

CONCLUSIONAG inhibited NO formation in dose-depended and time depended way. AG attenuated the changes of renal function induced by NO.

Animals ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; administration & dosage ; therapeutic use ; Female ; Guanidines ; administration & dosage ; therapeutic use ; Kidney Function Tests ; Male ; Nitric Oxide Synthase ; antagonists & inhibitors ; metabolism ; Rabbits ; Random Allocation ; Shock, Septic ; drug therapy ; Treatment Outcome

Connective tissue growth factor (CTGF) is a known profibrotic cytokine. The purpose of the present study was to explore the effects of inhalation of aminoguanidine (AG) on the up-regulation of CTGF in fibrotic lungs of rats. Sprague-Dawley rats received single intratracheal instillation of bleomycin (BLM) or instillation of the same volume of normal saline (NS) as control. From day 1 to day 30 after intratracheal BLM instillation, the rats inhaled AG (2, 10 or 50 mmol/L, 5 min each time) twice a day or inhaled the same volume of NS as vehicle control. The change of nitric oxide (NO) content in lungs was evaluated by nitrite/nitrate (NO₂(-)/NO₃(-)) content in out-flowing pulmonary plasma (OPP). The degree of fibrosis in lung was evaluated by the content of hydroxyproline (chloramine T method) and area of collagen (Masson stain) in lung. The CTGF expression in lung was detected by Western blot and RT-PCR. The contents of NO₂(-)/NO₃(-) were increased in OPP of rats on day 14 after the instillation of BLM, compared with those in the rats with instillation of NS [(156+/-21) mumol/L vs (51+/-15) mumol/L, P<0.01]. The content of hydroxyproline, the area of collagen, and the levels of CTGF protein and mRNA were increased in lungs of rats on day 30 after intratracheal instillation of BLM, compared with those in the rats with instillation of NS [hydroxyproline, (51+/-10) mg/g lung vs (20+/-5) mg/g lung; area of collagen, (38.7+/-8.8)% vs (5.7+/-1.5)%; CTGF protein, (1+/-0.25) vs (0.3+/-0.1); CTGF mRNA, (0.8+/-0.2) vs (0.15+/-0.03), P<0.01]. The above-mentioned indices were ameliorated by the inhalation of AG (10 or 50 mmol/L) (NO₂(-)/NO₃(-) content, P<0.01; other indices, P<0.05). It is therefore concluded that the inhalation of AG prevented the up-regulation of CTGF in fibrotic lungs of rats suffering from BLM instillation, which might be one of the mechanisms of the anti-fibrosis of AG in lungs.
Administration, Inhalation ; Animals ; Bleomycin ; adverse effects ; Connective Tissue Growth Factor ; metabolism ; Guanidines ; pharmacology ; Lung ; metabolism ; pathology ; Pulmonary Fibrosis ; drug therapy ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Up-Regulation

Although the incidence of bleeding complications during extracorporeal membrane oxygenator (ECMO) support has decreased in various trials, bleeding is still the most fatal complication. We investigated the ideal dosage and efficacy of nafamostat mesilate for use with ECMO in patients with acute cardiac or respiratory failure. We assessed 73 consecutive patients who received ECMO due to acute cardiac or respiratory failure between January 2006 and December 2009. To evaluate the efficacy of nafamostat mesilate, we divided the patients into 2 groups according to the anticoagulants used during ECMO support. All patients of nafamostat mesilate group were male with a mean age of 49.2 yr. Six, 3, 5, and 3 patients were diagnosed with acute myocardial infarction, cardiac arrest, septic shock, and acute respiratory distress syndrome, respectively. The mean dosage of nafamostat mesilate was 0.64 mg/kg/hr, and the mean duration of ECMO was 270.7 hr. The daily volume of transfused packed red blood cells, fresh frozen plasma, and cryoprecipitate and the number of complications related to hemorrhage and thrombosis was lower in the nafamostat mesilate group than in the heparin group. Nafamostat mesilate should be considered as an alternative anticoagulant to heparin to reduce bleeding complications during ECMO.
Acute Disease ; Anticoagulants/*administration & dosage ; Dose-Response Relationship, Drug ; *Extracorporeal Membrane Oxygenation ; Female ; Guanidines/*administration & dosage ; Heart Failure/diagnosis/mortality/therapy ; Heparin/administration & dosage ; Humans ; Male ; Middle Aged ; Myocardial Infarction/diagnosis/mortality/therapy ; Respiratory Distress Syndrome, Adult/diagnosis/mortality/therapy ; Retrospective Studies ; Shock, Septic/diagnosis/mortality/therapy ; Survival Analysis