Effects of Shengxue Pills on formaldehyde-induced cell damage in mice / 中草药

Objective: To explore the prophylactic and curative effects of Shengxue Pills on formaldehyde-induced cell damage of lung, liver, spleen, bone marrow, and peripheral blood lymphocytes in mice and the mechanisms. Methods: The mice were randomly divided into five groups: control, Shengxue Pill prophylaxis, Shengxue Pill therapy, and two model groups. ELISA was used to detect hydroxyl radicals, superoxide anion radical, and ornithine decarboxylase (ODC) activities in the lung, liver, and spleen cells. Bone marrow cells and peripheral blood lymphocyte micronucleus rate were observed with high power lens. Results: Hydroxyl radicals, superoxide anion radicals, and ODS activities in the cells of the organs in mice were investigated, and the number of bone marrow cells and peripheral blood lymphocyte micronucleus rate in mice in the prophylacitc and therapeutic groups were significantly greater than those in the model groups, respectively as well as in the normal group. Conclusion: Shengxue Pills could prevent and cure the formaldehyde-induced cell damages of the lung, liver, spleen, bone marrow, and peripheral blood lymphocytes of mice.

Antigout effect of renierol extracted from the marine sponge in the South China Sea / 中国海洋药物

Objective To investigate the inhibitive effect of renierol extracted from the marine sponge in the South China Sea on xanthine oxidase and the effect on mice hyperuricemia as well as its possible mechanisms.Methods Renierol was extracted from sponge and its antigoat effect was tested.The activity of xanthine oxidase was measured by using the generation of uric acid method and the NBT reduction induced by superoxide anions,then its possible mechanism was estimated.Results The two methods showed renierol could obviously inhibit the effect of xanthine oxidase with IC_(50)of 1.85 and 1.36?g?mL~(-1),respectively.Conclusion Renierol extracted from the marine sponge can effectively inhibit the effect of xanthine oxidase.

Effects of Nitric Oxide and Peroxynitrite Peroxynitrite on Sperm Motility and Viability / 대한비뇨기과학회지

PURPOSE: Recently, in assisted reproductive technologies(ART) programs theme Is an increasing Interest in the use of agents for the enhancement of sperm motility for assisted fertilization. In an attempt to improve the motility at the cryopreserved human semen and hence the fertilizing capacity of asthenospermic semen samples, different semen preparation techniques have been attempted and the effects of chemical stimulants as nitric oxide(NO) have been studied extensively. Superoxide anions cause lipid peroxidative damage to cell membrane phospholipids, and sperm are known to be particularly susceptible to lipid peroxidation. Such sperm with damaged membranes are impaired functionally. Recently, peroxynitrite, an anion and a potent oxidant, generated by the interaction of nitric oxide and superoxide anions has been demonstrated In macrophages and other cellular systems. Also this anion cause lipid peroxidative damage to cell membrane phospholiplds. We therefore Investigated whether NO and peroxynitrite have the roles to modulate sperm motility and to affect Its viability. MATERIALS AND METHODS: Normal human semen samples(as per World health Organization (WHO) criteria) were obtained after 3day period of abstinence by donors. The samples(n=5) were incubated with either sodium nitroprusside(SNP; 0.1, 05, 1 or 2mM) or peroxynitrite (10, 50 or 100 micrometer ) and the percent viability and motility were assessed at various time inteval up to 4hr. The human semen samples were treated with N-acetyl-L-cystein(NAC;10mM), SNP (0.5mM), phorbol myristate acetate(PMA; 100nM), of SNF plus PMA. Both superoxide and peroxynitrite release were measured directly by chemiluminometer. Percent viability and motility were assessed at 4hr of Incubation. A sample of each aliquot was placed in a Mauler chamber for videomicrography Percent motility were analyzed by using the sperm analysis imaging system. The sperm vlability was assessed by flow cytometer using LIVE/DEAD sperm viability kIt. The production of superoxide and peroxynitrite were measured by the method of chemiluminescence assay. Result : All results represent a mean +/-SEM, n=5. Treatment of human semen samples for 4hr with SNP, a NO generating agent, significantly decreased sperm motility and viability in high concentration [relative motility(% of control); 38 +/-4 and 30 +/-5, relative viability; 42 +/-4 and 30 +/-3 by 1 and 2mM of SNP]. In the presence of low concentration SNP(0.5mM), the sperm viability was not significantly affected(82 +/-3), whereas the sperm motility was affected(64 2). SNP(0.5mM) also decreased sperm motility(80 +/-2 at 2hr 64 +/-3 at 4hr, 44 +/-3 at 6hr, and 38 +/-4 at 8hr) in a time dependent manner. Since it was demonstrated that superoxide anions are one of the common source of lipid peroxidation, we investigated whether superoxide anions produced by human semens could Interact wlth NO to generate peroxynitrite. Adding N-acetyl-L-cystein(NAC) to the human semen samples partially blocked spontaneous release of superoxide, whereas PMA augmented the release of superoxide from human semen samples (control:0.9 106 0.3, NAC: 0.5 106 +/-0.4, and PMA: 2.5 106 +/-0.4photons/60min). The production of superoxide was corresponded with the production of peroxynltrite(control: 1.0 104CPM, SNP: 3.8 106CPM, SNP plus PMA. 12chi106CPM). In addition, SNP in combination with PMA(65 +/-3) markedly decreased sperm motility than that of SNP alone(77 +/-2.5) at 4hr, implying that nitric oxide might inhibit sperm motility via the formation of peroxynitrite In human semen samples. Exogenous peroxynitrite also decreased sperm motility in a dose dependent manner(10 micrometer : 64 +/-2, 50rM: 53 +/-3, and 1 0 micrometer of peroxynitrite: 23 4). CONCLUSIONS: These results suggest that NO inhibits sperm motility via the formation of peroxynltrite and further demonstrate that NO-induced inhibition of sperm motility is depended on the production of superoxide from human semens because peroxynitrite is generated by the interaction of NO and superoxide.

Inhibition of xathine oxidase by rosmarinic acid / 第二军医大学学报

Objective:To study the inhibitory effect of rosmarinic acid on xanthine oxidase.Methods: Xanthine oxidase((0.1 U/ml)) was incubated with xanthine(1 mmol/L for determining formation of uric acid;50 ?mol/L for determining superoxide anions) in the presence of 20,40 and 60 ?g/ml rosmarinic acid or allopurinol as positive control.The formation of uric acid was determined by automatic biochemical analyzer 5 min after reaction and the production of superoxide anions was measured by Nitro Blue Btetrazolium(NBT) reduction.HL-60 cells(1 ml,2?10~(5)/ml) were pretreated with xanthine(100 ?l,6 mol/L) and xanthine oxidase(100 ?l,0.1 U/ml),then rosmarinic acid(500 ?g/ml) or allopurinol(1 ?g/ml,as positive control)(AnnexinⅤ-PI kit) was added to determine the cell apoptosis rate.HL-60 cells(1 ml,2?10~(5)/ml) were also pretreated with xanthine(100 ?l,6 mol/L) and xanthine oxidase(100 ?l,0.1 U/ml),then rosmarinic acid(500 ?g/ml) or SOD(100 U/ml,as positive control)(cell cycle method) was added to determine the cell apoptosis rate.Results: Rosmarinic acid obviously inhibited the production of uric acid and superoxide anion-induced reaction in NBT assay,with their IC_(50)being 56 ?g/ml and 21 ?g/ml,respectively.The rates of apoptosis inhibition by rosmarinic acid were both over 40% by Annexin Ⅴ-PI kit and cell cycle method.Conclusion: Rosmarinic acid is a competitive inhibitor of xanthine oxidase.