Considering that high levels of nitric oxide (NO) exert anti-cancer effect and the derivatives of oleanolic acid (OA) have shown potent anti-cancer activity, new O-vinyl diazeniumdiolate-based NO releasing derivatives (5a-l, 11a-l) of OA were designed, synthesized, and biologically evaluated in the present study. These derivatives could release different amounts of NO in liver cells. Among them, 5d, 5i, 5j, 11g, 11h, and 11j released more NO in SMMC-7721 cells and displayed stronger proliferative inhibition against SMMC-7721 and HepG2 cells than OA and other tested compounds. The most active compound 5j showed almost 20-fold better solubility than OA in aqueous solution, released larger amounts of NO in liver cancer cells than that in normal ones, and exhibited potent anti-hepatocellular carcinoma activity but little effect on the normal liver cells. The inhibitory activity against the cancer cells was significantly diminished upon addition of an NO scavenger, suggesting that NO may contribute, at least in part, to the activity of 5j.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Azo Compounds ; chemistry ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drug Screening Assays, Antitumor ; Hep G2 Cells ; Hepatocytes ; drug effects ; metabolism ; pathology ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; Nitric Oxide ; chemistry ; Nitric Oxide Donors ; chemical synthesis ; chemistry ; pharmacology ; Oleanolic Acid ; analogs & derivatives ; chemistry ; pharmacology

BACKGROUND/AIMS: Ozone is an environmentally reactive oxidant, and pycnogenol is a mixture of flavonoid compounds extracted from pine tree bark that have antioxidant activity. We investigated the effects of pycnogenol on reactive nitrogen species, antioxidant responses, and airway responsiveness in BALB/c mice exposed to ozone. METHODS: Antioxidant levels were determined using high performance liquid chromatography with electrochemical detection. Nitric oxide (NO) metabolites in bronchoalveolar lavage (BAL) fluid from BALB/c mice in filtered air and 2 ppm ozone with pycnogenol pretreatment before ozone exposure (n = 6) were quantified colorimetrically using the Griess reaction. RESULTS: Uric acid and ascorbic acid concentrations were significantly higher in BAL fluid following pretreatment with pycnogenol, whereas gamma-tocopherol concentrations were higher in the ozone exposed group but were similar in the ozone and pycnogenol pretreatment groups. Retinol and gamma-tocopherol concentrations tended to increase in the ozone exposure group but were similar in the ozone and pycnogenol pretreatment groups following ozone exposure. Malonylaldehyde concentrations increased in the ozone exposure group but were similar in the ozone and pycnogenol plus ozone groups. The nitrite and total NO metabolite concentrations in BAL fluid, which parallel the in vivo generation of NO in the airways, were significantly greater in the ozone exposed group than the group exposed to filtered air, but decreased with pycnogenol pretreatment. CONCLUSIONS: Pycnogenol may increase levels of antioxidant enzymes and decrease levels of nitrogen species, suggesting that antioxidants minimize the effects of acute ozone exposure via a protective mechanism.
Animals ; Antioxidants/*pharmacology ; Ascorbic Acid/metabolism ; Bronchial Hyperreactivity/chemically induced/metabolism/*prevention & control ; Bronchoalveolar Lavage Fluid/chemistry ; Bronchoconstriction/drug effects ; Disease Models, Animal ; Female ; Flavonoids/*pharmacology ; Inhalation Exposure ; Lung/*drug effects/enzymology/physiopathology ; Malondialdehyde/metabolism ; Mice ; Mice, Inbred BALB C ; Nitric Oxide/metabolism ; Oxidative Stress/*drug effects ; *Ozone ; Uric Acid/metabolism ; Vitamin A/metabolism ; alpha-Tocopherol/metabolism

AIM: To investigate the antioxidant and anti-inflammatory effects of Shengmai San (SMS) and its ethyl acetate extract (SEa), n-butanol extract (SBu), and aqueous extract (SWe), and clarify the material base of SMS and the roles played by its fractions. METHODS: A mouse model of transient forebrain ischemia/reperfusion (I/R) by means of common carotid artery occlusion (CCAO) was used to investigate the effects of SMS and its three fractions. Histopathological damage, blood-brain barrier disruption, and antioxidant and inflammation-related parameters, including malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), myeloperoxidase (MPO), nitric oxide (NO), tumor necrosis factor-α (TNF-α) were measured. The chemical constituents of each fraction were identified by LC-MS. RESULTS: Eighteen lignans in SEa, and thirteen steroidal glycosides and ginsenosides in SBu were determined. SMS significantly inhibited I/R induced formation of histological injury and cerebral MPO activity. SMS showed the strongest antioxidant and anti-inflammatory effects against the I/R-caused injuries. SEa showed higher antioxidant activity than the other two fractions and SBu has a slightly stronger inhibition on the productions of NO and TNF-α. CONCLUSION SMS as a whole had the most effective protection against cerebral I/R-caused injuries compared with its fractions, which inferred that it contains different groups of compounds that contribute together to its protective effect.
Animals ; Chromatography, High Pressure Liquid ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Glutathione Peroxidase ; genetics ; metabolism ; Humans ; Male ; Malondialdehyde ; metabolism ; Nitric Acid ; metabolism ; Oxidative Stress ; drug effects ; Peroxidase ; genetics ; metabolism ; Protective Agents ; administration & dosage ; chemistry ; Rats ; Reperfusion Injury ; drug therapy ; genetics ; metabolism ; prevention & control ; Superoxide Dismutase ; genetics ; metabolism

OBJECTIVETo investigate the effects of caffeic acid ester fraction (Caf) from Erigeron breviscapus, mainly composed of dicaffeoylquinic acids (diCQAs), on microglial activation in vitro and focal cerebral ischemia in vivo.

METHODSThe production of nitric oxide (NO), tumor necrosis factor α (TNF-α), and interleukin-1β (IL-1β) induced by lipopolysaccharide (LPS) treatment in rat primary cultured microglia were measured by Griess reaction or enzyme-linked immunosorbent assay. Cell viability of cortical neurons was measured using AlamarBlue reagent. The behavioral tests and the infarct area of brain were used to evaluate the damage to central nervous system in rat middle cerebral artery occlusion (MCAO) model of cerebral ischemia. Real time polymerase chain reaction was used to determine the expression of inducible nitric oxide synthase (iNOS), TNF-α and IL-1β mRNA in ischemic cerebral tissues.

RESULTSCaf inhibited the production of NO, TNF-α and IL-1β induced by LPS treatment in primary microglia in a dose-dependent manner. Exposure of cortical neurons to conditioned medium from Caf-treated microglia increased neuronal cell viability (P<0.01) compared with conditioned medium from LPS-treated alone. In MCAO rat model of cerebral ischemia, Caf could significantly improve neurobehavioural performance and reduce percentage infarct volume compared with the vehicle group (P<0.05). Caf could also significantly inhibit the up-regulation of iNOS, TNF-α, and IL-1β gene expressions in ischemic cerebral tissues.

CONCLUSIONCaf could suppress microglial activation, which may be one mechanism of its neuroprotective effect against ischemia.

Animals ; Brain ; drug effects ; metabolism ; pathology ; Brain Ischemia ; complications ; drug therapy ; pathology ; prevention & control ; Caffeic Acids ; chemistry ; pharmacology ; Chemical Fractionation ; Chromatography, High Pressure Liquid ; Erigeron ; chemistry ; Gene Expression Regulation ; drug effects ; Infarction, Middle Cerebral Artery ; complications ; pathology ; Interleukin-1beta ; genetics ; metabolism ; Microglia ; drug effects ; metabolism ; pathology ; Neuroprotective Agents ; chemistry ; pharmacology ; therapeutic use ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Plant Extracts ; pharmacology ; Quinic Acid ; analogs & derivatives ; chemistry ; pharmacology ; therapeutic use ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

OBJECTIVETo study and compare the changes of toxicity of Euphorbia pekinensis, E. kansui and E. ebracteolata before and after being prepared by vinegar.

METHODSmall intestinal accentuation of mice and peritoneal macrophage NO release experiments were assessed to investigate the changes of toxicity of the three Chinese Medicines of Euphorbia before and after being prepared.

RESULTE. pekinensis, E. kansui and E. ebracteolata and vinegar can obviously promot small intestinal accentuation and peritoneal macrophage NO release with the intensity of toxicity in the order of E. kansui > E. pekinensis > E. ebracteolata. After being prepared with vinegar, the toxicity of the three medicines decreased obviously compared to crude one.

CONCLUSIONE. pekinensis, E. kansui and E. ebracteolata can induce inflammation and accelerate enterokinesis. After being prepared with vinegar, the irritation on Euphorbia decreased obviously.

Acetic Acid ; chemistry ; Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; toxicity ; Euphorbia ; chemistry ; toxicity ; Female ; Intestine, Small ; drug effects ; metabolism ; Macrophages, Peritoneal ; drug effects ; metabolism ; Male ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred ICR ; Nitric Oxide ; analysis ; metabolism ; Plant Roots ; chemistry ; toxicity ; Toxicity Tests

To find novel antihepatitis drugs, a series of nitrate-oleanolic acid (OA) hybrids (10a, 10b, 11a-11e and 12a-12c) were designed and synthesized on the basis of previous studies using OA as lead compound, which is widely found in natural plants and liver-specific metabolism. In the present study, ten novel NO-releasing derivatives of OA were synthesized by connecting nitrate to the OA-3-OH through varying lengths of linkers containing antioxidants which were designed to increase the ability of these target compounds to scavenge free radicals. The structures of these objective compounds were determined by IR, MS, 1H NMR and elemental analysis. Their protective effects on anti-Fas mediated HepG2 cell apoptosis were in vitro evaluated by LDH assay. Compound 12a is the most potent inhibitor. Its effect on anti-Fas mediated HepG2 cell apoptosis and amount of NO-releasing in vitro are similar to those of positive control NCX-1000.
Antioxidants ; chemistry ; Apoptosis ; drug effects ; Hep G2 Cells ; Humans ; Nitrates ; chemical synthesis ; chemistry ; pharmacology ; Nitric Oxide ; metabolism ; Nitric Oxide Donors ; chemistry ; Oleanolic Acid ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship ; Ursodeoxycholic Acid ; analogs & derivatives ; pharmacology

OBJECTIVE: To investigate the advantages of nitric acid digestion method and its differences with the traditional method. METHODS: Ethanol was used to fully fix the testing sample. About 80-100 g of the testing samples were cut into pieces and digested with nitric acid. It was then centrifuged and washed to remove organic components. Smears were prepared and examined under the light microscope. RESULTS: The diatom had been identified with clear striations, counted conveniently and classified easily. CONCLUSION The improved nitric acid digestion method is not only simple with a higher successful rate of detection, but also can prevent interference from contamination. It can improve the stability of the experimental results, avoid harm to human and environment, and provide higher safety in the course of experiment.
Autopsy ; Diatoms/isolation & purification* ; Drowning ; Forensic Pathology ; Humans ; Kidney/metabolism* ; Liver/metabolism* ; Lung/metabolism* ; Nitric Acid/chemistry* ; Postmortem Changes ; Tissue Fixation/methods*

OBJECTIVETo observe the influence of intra-articular injection of sodium hyaluronate (SH) on the expression of inducible nitric oxide synthase (iNOS) in the synovium and nitric oxide (NO) content in synovial fluid of rabbits with traumatic osteoarthritis (OA).

METHODSSixteen white rabbits underwent unilateral anterior cruciate ligament transection and were randomly divided into 2 groups 5 weeks after the operation. Rabbits in the experimental group received intra-articular injection of 0.3 ml of 1% SH, once a week for 5 weeks. Animals in the control group were treated under the same conditions using physiological saline. All the animals were sacrificed at the 10th week after surgery. The mRNA expression of iNOS in the synovium was analyzed using reverse transcription-polymerase chain reaction. The content of NO in the synovial fluid was assayed.

RESULTSThe level of iNOS expression of the synovium in the experimental group was lower than that in control group (0.47+/-0.09 vs. 0.65+/-0.12, t equal to 3.45, P less than 0.01). Compared with control group, the content of NO decreased significantly in synovial fluid of SH injection group (134.11 micromolar/L +/- 12.47 micromolar/L vs. 152.17 micromolar/L +/- 15.69 micromolar/L, t equal to 2.55, P less than 0.05).

CONCLUSIONSSH significantly decreases the content of NO in the synovial fluid of rabbits with traumatic OA. SH may exert the effect on synovial fluid NO level as a result of the suppression of iNOS expression in the synovium. It may be one of the mechanisms of the therapeutic effect of SH on early traumatic OA.

Animals ; Anterior Cruciate Ligament Injuries ; Hyaluronic Acid ; pharmacology ; therapeutic use ; Nitric Oxide ; analysis ; Nitric Oxide Synthase Type II ; analysis ; Osteoarthritis ; drug therapy ; etiology ; metabolism ; Rabbits ; Random Allocation ; Synovial Fluid ; chemistry ; Synovial Membrane ; enzymology

OBJECTIVETo observe the effects of sapindoside on blood pressure, Ang II, Ald, ET in the blood plasma and NO in the serum in renovascular hypertension rat.

METHODThe 2K1C (2 kidney 1 clap) hypertensive model rats were used and drugs had been given by ig. for 5 weeks. The blood pressure was measured at the 1, 3, 7, 14, 21, 28, 35 day after drug. At the end of 5th weeks, the Ang II, Ald, ET in the blood plasma and NO in the serum were measured.

RESULTSapindoside (H, M and L) by ig. for 5 weeks could significantly lower the blood pressure, increase the levels of NO in the serum, reduce the concentration of Ang II, Ald, ET in the blood plasma.

CONCLUSIONSapindoside plays an important role in decreasing the blood pressure of renovascular hypertension rat.

Aldosterone ; blood ; Angiotensin II ; blood ; Animals ; Antihypertensive Agents ; isolation & purification ; pharmacology ; Blood Pressure ; drug effects ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Endothelins ; blood ; Female ; Hypertension, Renovascular ; blood ; physiopathology ; Male ; Nitric Oxide ; blood ; Oleanolic Acid ; analogs & derivatives ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sapindus ; chemistry ; Saponins ; isolation & purification ; pharmacology

OBJECTIVETo investigate 7 gene expression profile associated with inflammation and oxidative stress in vascular endothelium injure of rats with deficiency of vital energy or qi stagnation, and the effect of Tongxinluo on gene expression profile.

METHODThe model of vascular endothelium injury of rats with deficiency of vital energy or qi stagnation were established by using high L-methionine, with load-carrying swimming or being fastened, respectively. RT-PCR and SAGE database which is available in NCBI, were used to analyze the changes of 7 gene expression related with inflammation and oxidative stress in endothelium injure and the effect of Tongxinluo on the gene expression profile.

RESULTCompared with control group, the gene expression of inflammation related COX-1, COX-2, oxidative stress related iNOS, SOD and blood vessel vasomotion related eNOS, ECE, increased in deficiency of vital energy group (P < 0.05 or P < 0.01), and the gene expression decreased with Tongxinluo treatment (P < 0.05 or P < 0.01). The gene expression of COX-1, COX-2, iNOS and eNOS, ECE, increased (P < 0.01), but the gene expression of PCS and SOD decreased (P < 0.01), in qi stagnation group, and the disorder of gene expression improved with treatment of Tongxinluo (P < 0.01).

CONCLUSIONThe 7 gene expression related to vascular endothelium injure were not the same in rat with deficiency of vital energy or qi stagnation, and Tongxinluo could regulate the disorder of the gene expression, protecting vascular endothelium from injure.

Animals ; Aspartic Acid Endopeptidases ; genetics ; Cyclooxygenase 1 ; genetics ; Cyclooxygenase 2 ; genetics ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Endothelin-Converting Enzymes ; Endothelium, Vascular ; injuries ; physiopathology ; Gene Expression ; drug effects ; Gene Expression Profiling ; Male ; Medicine, Chinese Traditional ; Metalloendopeptidases ; genetics ; Nitric Oxide Synthase Type II ; genetics ; Nitric Oxide Synthase Type III ; genetics ; metabolism ; Qi ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Superoxide Dismutase ; genetics