Effects of sodium ferulate on the nonenzymatic glycation and oxidation in kidneys of diabetic rats

AIM: To investigate the blocking effects of sodium ferulate (SF) on the nonenzymatic glycation and oxidation in kidneys of diabetic rats. METHODS: The diabetic rats induced by streptozotocin (STZ) were treated with SF (110 mg/kg) per day for 8 weeks. The renal weight/body weight, clearance rate of creatinine (Ccr), proteinuria, advanced glycation end products (AGEs) in renal cortex, and the malonyldialdehyde (MDA), fructosamine (FMN), antioxidant enzymes in serum and renal cortex were measured. The pathologic changes of kidneys were also observed. RESULTS: The levels of Ccr, proteinuria, FMN and AGEs in renal cortex, FMN in serum, and renal weight/body weight in diabetic control group were significantly higher than those in normal control group. In the diabetic control group, there was a highter level of MDA and a lower activity of superoxide dismutase (SOD) and catalase (CAT) in serum and renal cortex than those in normal control group. Except for FMN in renal cortex, the abnormalities were significantly ameliorated in the treatment group. The pathologic changes was significant in the diabetic control group, which was significantly ameliorated with the treatment of SF. CONCLUSIONS: SF protects the activity of antioxidant enzymes, clears away oxygen radicals and inhibits the deposit of AGEs in kindey, which may be the mechanisms of protecting effects of SF on kindey in diabetic rats.     

Effect of sodium ferulate on bcl-2 and bax gene expression of apoptotic hippocampal neurons induced by sodium ferulate

AIM: To investigate the protective effects of sodium ferulate (SF) on apoptosis in cultured hippocampal neurons induced by sodium nitroprusside (SNP), and the effect of SF on expression of bcl-2 and bax. METHODS: The primary cultured hippocampal neurons were exposed to 50 μmol SNP, a nitric oxide-donor, for 24 h after pretreatment with different concentrations of SF (10-160 μmol/mL) for 6 h. Then neuronal viability was tested by MTT assay. Fluorescent staining with Hoechst 33258 and agarose gel electrophoresis was used to analyze apoptosis. The expressions of bcl-2, bax mRNA and protein were tested by RT-PCR and Western blotting. RESULTS: Pretreatment with SF(10-160 μmol/L) for 6 h increased the survival rate of neurons. SF prevented the neuronal nuclei from shrinkage, condensation and cleavage and blocked neuronal nuclear DNA fragmentation induced by SNP. SF also increased the expressions of bcl-2 mRNA and Bcl-2 protein and decreased the expressions of bax mRNA and Bax protein. CONCLUSION: SF prevents the cultured hippocampal neurons against SNP neurotoxicity. The mechanism of protection is related to the increase in Bcl-2 level and the decrease in Bax level. As a result, the ratio of Bcl-2/Bax is changed.     

Effects of sodium ferulate on function of macrophages in the colonic tissue of colitis rats

AIM: To explore the effects of sodium ferulate (SF) on function of macrophages in colonic tissue of the colitis rats in vivo. METHODS: The immunological colitis model of rats was produced. SF was used intracolonically for 21 days. The contents of malondialdehyde (MDA), nitric oxide (NO), prostaglandin E₂ (PGE₂) and the activity of superoxide dismutase (SOD), interleukin-1 (IL-1), TNF-α, myelopexoxidase (MPO), and the expression level of NF-κB p65 in colonic tissue of the rats were detected. RESULTS: SF (200,400,800 mg/kg) decreased the elevated contents of MDA, NO, PGE₂, the activity of IL-1, TNF-α, MPO, and the expression level of NF-κB p65, while increased the reduced activity of SOD in colonic tissue of the colitis rats in a dose-depended manner. CONCLUSION: SF restrained the activity of activated colonic macrophages and relieved the colonic inflammation reaction in vivo in colitis rats, which may be related to the suppression of NF-κB activation.     

Effect of sodium ferulate on expression of BCl-2 and bax in rat lens epithelial cells

AIM: To investigate the effect of sodium ferulate on expression of apoptosis-related genes, BCl-2 and bax, in rat lens epithelial cells (LEC) injured by oxidation.METHODS: Eyes of SD rats were divided randomly into four groups: control group, hydrogen peroxide group (H₂O₂), pirenoxine sodium group (PS) and sodium ferulate group (SF). Eyes were excised and lenses were separated under operating microscope and sterilized condition. Lenses were incubated in CO₂ incubator for 24 h with 300μmol·L^-1H₂O₂ and with or rithout 5 mmol·L^-1SF. The expression of BCl-2 and Bax protein of LEC were measured and compared by tearing the LEC anterior capsule via immunohistochemical analysis.RESULTS: (1) There were BCl-2 and Bax expression in normal lenses of SD rates, BCl-2 expression was stronger than Bax. (2) BCl-2 expression decreased and Bax expression increased markedly (P<0.01),BCl-2/Bax reduced in H₂O₂ group. (3) There were up regulation of BCl-2 expression and down-regulation down of Bax expression, and BCl-2/Bax increased in SF group compared with H₂O₂ group (P<0.01). (4) The above changes were similar between PS group and SF group, but much stronger in SF group than that in PS group (P<0.01).CONCLUSION: These results show that SF regulates expression of apoptosis-related genes, BCl-2 and bax, which may be the molecular mechanism of LEC apoptosis inhibition by SF.     

Effect of sodium ferulate on heat shock protein 27 phosphorylation in vascular smooth muscle cells

AIM: To investigate the effect of sodium ferulate on phosphorylation of heat shock protein 27 (HSP27) in vascular smooth muscle cells (VSMCs) induced by angiotensin II (AngII) and platelet derived growth factor-BB (PDGF-BB).METHODS: Cultured VSMCs derived from rat thoracic aorta were used.The activity of HSP27 was evaluated by Western blotting with specific phospho-HSP27 antibody.RESULTS: The phosphorylation of HSP27 in response to AngII and PDGF-BB was suppressed by sodium ferulate in a dose-dependent manner,with maximal inhibition rates of 39.0% (P<0.05) and 56.8% (P<0.01) respectively at concentration of 10-4 mol/L.CONCLUSION: HSP27 phosphorylation induced by AngII and PDGF-BB in VSMCs may be significantly inhibited by sodium ferulate.     

Effects of ligustrazine ferulate on myocardial apoptosis and apoptosis-related protein expression in rats with myocardial ischemia-reperfusion injury

AIM:To observe the effects of ligustrazine ferulate on the apoptosis of myocardial cells in rats with myocardial ischemia-reperfusion injury, and to explore its possible mechanism. METHODS:Sixty male SD rats were randomly divided into five groups: sham-operation group, ischemia-reperfusion group, ligustrazine (4 mg/kg) group, low-dose (4 mg/kg) ligustrazine ferulate group and high-dose (8 mg/kg) ligustrazine ferulate group. The rat myocardial ischemia-reperfusion model was established by 30 min of myocardial ischemia followed by 120 min of reperfusion. Drugs were administered to the rats by jugular vein injection 10 min before reperfusion. After the reperfusion was finished, the biochemical indicators in serum and the histological indexes in myocardium were detected. RESULTS: Compared with ischemia-reperfusion group, ligustrazine ferulate lowered the serum levels of creatine kinase MB form, lactate dehydrogenase, cardiac troponin I and malondialdehyde, elevated the activity of total superoxide dismutase in serum and the expression of Bcl-2 protein in myocardium, decreased the expression of Bax protein and myocardial apoptotic index, and increased the Bcl-2/Bax ratio (all P<0.01). All the indicators in ligustrazine ferulate groups were dose-dependently superior to those in ligustrazine group (P<0.05 or P<0.01). CONCLUSION: Ligustrazine ferulate protects rats against myocardial ischemia-reperfusion injury. Its anti-apoptotic effect may be related to up-regulation of Bcl-2 and down-regulation of Bax.     

Inhibitory effects of Angelicae sinensis and sodium ferulate on acute inflammatory liver injury and expression of ICAM-1 and E-selectin in mice

AIM: To explore the effects of Angelicae sinensis preparation and sodium ferulate on inflammatory liver injury induced by lipopolysaccharide (LPS) and their molecule mechanism. METHODS: ICR mice were divided into five groups: Angelicae sinensis group, sodium ferulate group, dexamethasone group, inflammation control group and normal group. The model of inflammatory injury in mice was set up by tail vein injection with the bacillus calmette-guerin (BCG) and LPS respectively prior and posterior to administration of those tested drugs. The tested drugs (the preparations of Angelica sinensis, sodium ferulate and dexamethasone) and normal saline were given respectively to the corresponding group. The pathological observation of the liver tissues in mice was made for the intensity of inflammatory liver injuries. The immunohistochemical detection and comparison were performed for the expression of ICAM-1 and E-selectin proteins in the liver tissues in those mice. RESULTS: The intensities of liver inflammatory injuries in mice from drug-treated groups were obviously lighter than that from the inflammation control group (P<0.01). The expressions of ICAM-1 and E-selectin proteins in the liver tissues of mice from the inflammation control group were not only significantly higher than that in normal control, but also obviously higher than that from drug-treated groups (P<0.01). CONCLUSIONS: Angelica sinensis and sodium ferulate alleviate inflammatory liver injury induced by injection of LPS. Their suppressive effects on inflammatory liver injury may be associated with the decrease in the expression of ICAM-1 and E-selectin proteins.     

Effects of Taoren Honghua Jian on the hyperlipidemia without symptom

AIM: To examine the effect of traditional chinese medicine recipe, Taoren Honghua(semen persicae-flos carthami) decoction, on hyperlipidemia without symptom. METHODS: The plasma TC, TG, LDL, HDL, apolipoprotein(Apo) A, Apo Bof the patients with hyperlipidemia without symptom were measured using automatic analyzer (shimadiu CL-7200), the production of nitric oxide(NO) was detected by Greiss reaction, and SODactivity and MDAformation were examined using o-trihydroxy benzene and barbituric methotheds, respectively. RESULTS: After oral administration of Taoren Honghua decoction, the plasma levels of TC, TG, LDL, and MDAof the patients were markedly decreased, however, the plasma levels of ApoA, HDL, SODand NOwere significantly increased and almost no change was detected in the plasma level of Apo B. In control group, it was found that although the plasma level of TC, TG and LDL were decreased ( P<0.05 ) and ApoAas well as HDLwere increased, ApoB, SOD, MDA and NO production were all unchanged. CONCLUSION: The traditional chinese medicine recipe, Taoren Honghua decoction,has a significant therapeutic effect on patients with hyperlipidemia by removing blood stasis, promoting qi circulation and in turn reducing blood lipid level.     

Effects of ibrolipim against atherosclerosis in minipigs

AIM: To explore the application mechanism of NO-1886 (ibrolipim), a synthetic compound, improving dyslipidemia and inhibiting atherosclerosis in Guizhou minipigs fed with high fat/high sucrose diet. METHODS: Fifteen Chinese Guizhou minipigs were randomized into three groups with similar body weight ［(n=5 in normal control group (CD); n=5 in high fat/high sucrose group (HFSD); n=5 in high fat/high sucrose supplemented ibrolipim group （HFSD+ibrolipim)］. Blood samples were withdrawn from the eyehole sinus venosus of the animals at the end of each month after fasting overnight. The animals were sacrificed at the end of 8 months. The concentrations of cholesterol ester in plasma HDL were analyzed by HPLC. The aortic fatty streak-lesions were quantified following lipid staining with Sudan IV. Lipid droplets in liver were observed by Oil red O staining. RESULTS: Compared with CD, fasting plasma TC, TG and FFA levels of HFSD were elevated significantly. The aortic fatty streak-lesions were clearly presented in the animals’ aortas. The intima became rougher and thicker. A lot of lipoid foam cells migrated to regions of intima and smooth muscle cells, which associated with the injuries of internal elastic lamina. Extensive fat deposited in the liver were observed. Supplementing of 1.0% ibrolipim into high fat/high sucrose diet induced the decrease in plasma TG and FFA concentrations and an increase in plasma HDL-C concentration compared with HFSD. A little fat deposited in the liver were observed. CONCLUSION: ibrolipim prevents AS in high fat/high sucrose diet feeding minipigs through decreasing the plasma TG and elevating the plasma HDL-C.     

Inhibitory effect of sodium ferulate on Aβ25-35-induced p38 MAPK activation

AIM: To investigate effect of sodium ferulate on Aβ_25-35-mediated signaling pathway. METHODS:The isolated peritoneal macrophages from mice were cultured. p38 MAPK protein kinase in nuclear extracts was analyzed by Western blotting. The concentration of TNF-α and NO in supernatant were measured by ELISA and Griess reaction technique. The expression of iNOS protein was detected by immunochemical technique. RESULTS:Aβ_25-35 significantly increased the concentrations of TNF-α and NO in supernatant, expression of iNOS in macrophages and p38 MAPK protein kinase in nuclear extracts, which were blocked by sodium ferulate. CONCLUSION:Sodium ferulate inhibits p38 MAPK activation triggered by Aβ_25-35.     

Effects of Retinervus luffae fructus on serum lipid level in experimental hyperlipidemia rats

AIM: To investigate effects of Retinervus luffae fructus (RLF) on level of serum lipid and body weight in hyperlipidemia rats. METHODS: Thirty-two male SD rats were randomly divided into four groups: control group (A), hyperlipidemia group (B), hyperlipidemia + RLF group (C), RLF group (D). Both group A and C were fed normal diet every day, while group B and group D fed high fat diet. Meanwhile, group C and D were administered with RLF solution at the dose of 10 mL/kg, respectively for 14 days, while group A and B were administered with drinking water. RESULTS: (1) At the end of experiment, a significant reduction was found in the levels of serum total cholesterol (TC) and triglyceride (TG) of group C animals treated with RLF solution; (2) The levels of serum TC of group D was progressively decreased compared to the level of serum TC at the beginning of experiment; (3) The level of high-density lipoprotein cholesterol (HDL-C) of group C remained unaltered 8d after treatment with RLF solution; (4) The body weight in group C was obviously lower than that in group B. CONCLUSION: RLF had an obvious hypolipidemic effect on hyperlipidemia rats. It can inhibit the decrease in the HDL-C and the increase of body weight in rats.     

Effects of Sini decoction on atherosclerosis and ceramide content of aorta in rabbits

AIM:To observe the effects of Sini decoction on atherosclerosis(AS) and ceramide content of aorta in rabbits. METHODS:28 rabbits were randomly divided into three groups. Control group was fed with a normal diet; High cholesterol group was fed 1% cholesterol and 5% fat diet; Sini decotion+ high cholesterol group was fed 1% cholesterol and 5% fat diet plus Sini decotion (4.2 g·kg^-1·d^-1). At the end of study, the plaque area were measured, the atorta ceramide and cell apoptosis were also detected. RESULTS:Sini decotion diminished lipid plaque area on the aortic endothelium, reduced the levels of aorta ceramide and the apoptosis index. CONCLUSION:Sini decoction has an inhibitory effect on AS, the mechanism may be that Sini decotion reduces concentration of ceramide in aorta.     

Effects of pioglitazone on myocardial energy metabolism and hemodynamics in rats with myocardial infarction

AIM: To observe the effects of pioglitazone on myocardial energy metabolism and hemodynamics in rats with heart failure after myocardial infarction (MI). METHODS: The model of MI was established by ligation of left anterior descending artery. The 20 surviving rats were randomly divided into MI group (n=10) and pioglitazone intervention group (P group,n=10, pioglitazone 3 mg·kg^-1·d^-1 orally). The sham-operated rats (SH, n=10) served as controls. Hemodynamic parameters were measured. The ratio of left ventricular weight to body weight (LVW/BW) and the ratio of right ventricular weight to body weight (RVW/BW) were calculated after 8-week treatment. The expression of PPARγ was examined by Western blotting. Mitochondrial respiratory function was determined by Clark oxygen electrodes. The size of adenine acid pool (ATP, ADP and AMP) in mitochondria was measured by HPLC. The adenine nucleotide translocator(ANT) activity was detected by the atractyloside-inhibitor stop technique. RESULTS: Compared with SH group, the protein expression of PPARγ was significantly decreased in MI group (P<0.01). The mitochondrial respiratory activity, the transport activity of ANT and the high-energy phosphate content were decreased in MI group (P<0.01), and the hemodynamic parameters were in disorder (P<0.01). Compared with MI group, the protein expression of PPARγ in P group was significantly increased. The mitochondrial respiratory activity, the high-energy phosphate content, the transport activity of ANT were improved (P<0.01). However, the hemodynamic parameters were not significantly changed.CONCLUSION: Pioglitazone increases the protein expression of PPARγ and improves myocardial energy metabolism in the development of heart failure in the rat model of myocardial infarction, but dose not change the hemodynamic parameters significantly.     

Effects of Retinervus luffae fructus on mRNA expression of low-density lipoprotein receptor in hyperlipidemia mice

AIM: To observe the effects of Retinervus luffae fructus (RLF) on mRNA expression of low-density lipoprotein receptor (LDL-R) in hyperlipidemia mice. METHODS: Mice were fed with high fat diet to induce a hyperlipidemia model. By using xuezikang, a Chinese medicine, as a positive control, the effect of RLF on serum total cholesterol (TC) and level of low density lipoprotein cholesterol (LDL-C) in mice were observed. The liver total RNA was extracted by Trizol method. The LDL-R mRNA expression was determined by RT-PCR. RESULTS: (1) The levels of TC ［(5.71±0.82) mmol/L］ and LDL-C ［(3.99±1.12) mmol/L］ in hyperlipidemia (HPL) group were higher than those in control (P<0.01). The levels of TC ［(3.65±0.28) mmol/L］ and LDL-C ［(2.74±0.54) mmol/L］ in RLF treatment group, and the levels of TC ［(3.94±0.65) mmol/L］ and LDL-C ［(3.00±0.23) mmol/L］ in positive control (PC) group were lower than those in HPL group (P<0.01). (2) The level of hepatic LDL-R mRNA expression was lower in HPL group than that in control group (P<0.01). Compared to HPL group, significant increases in hepatic LDL-R mRNA expression in RLF treatment group and PC group (P<0.01) were observed. CONCLUSION: Retinervus Luffae Fructus exerts obviously lipid-lowering effect and enhances the hepatic LDL-R mRNA expression in experimental hyperlipidemia mice.     

Effects of Tongxinluo on activation of platelet in rabbit model of atherosclerosis

AIM: To study the effects of Tongxinluo on the activation of platelets in a rabbit model of atherosclerosis. METHODS: New Zealand rabbits were randomly divided into 7 groups: normal group, model group, the groups treated with high, medium and low doses of Tongxinluo micropowder (0.15, 0.3 and 0.6 g·kg^-1·d^-1), atorvastatin group (2.5 mg·kg^-1·d^-1), and aspirin group (12.5 mg·kg^-1·d^-1). The rabbits in normal group was fed with common diet for 12 weeks, and the rabbits in model group were fed with high-fat diet for 12 weeks to establish atherosclerosis model. The rabbits in the rest groups were treated with the corresponding drugs, at the same time to give high-fat diet. Fasting for 12 h after the last treatment, whole blood was collected to perform the blood routine test, and to measure serum and plasma levels of lipids, platelet factor 4 (PF4) and soluble CD62P (sCD62P). Flow cytometry was used to analyze platelet calcium ion concentration. Electron microscopy was used for platelet superfine observations, and light microscopy for observing the pathological changes. RESULTS: Compared with normal group, the levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), platelet counts, and mean platelet volume in model group were significantly elevated, and the levels of PF4, sCD62P and calcium were also significantly increased (P < 0.05). Compared with model group, except aspirin group, the levels of TG, TC and LDL-C in high, medium and low doses of Tongxinluo groups and atorvastatin group were effectively decreased. The platelet counts and mean platelet volume in all treatment groups were markedly decreased, and the serum levels of PF4, sCD62P and Ca²⁺ in platelet (P < 0.05) were reduced. In electron microscopic observation, the shape of platelet was regular and organelles distributed uniform in normal group. However, in model group, the shape of platelet was irregular, pseudopodia forming was obviously observed, and α particles and dense granules decreased, indicating that the platelet was activated. To a different extent, the platelet shape, increase in the number of α particles and dense granules were improved in treatment groups and the damage of the cytoplasm was attenuated. Through histopathological observation, the intimal was smooth and complete in normal group. In the model group, the intimal thickness markedly increased, foam cell aggregated, and plaque was formed. Compared with model group, the intimal thickening and the number of foam cells were significantly decreased, and plaque formation was not obvious in atorvastatin group and high dose of Tongxinluo group. The pathological damages in the other treatment groups were alleviated in different degrees. CONCLUSION: Tongxinluo significantly inhibits the activation of platelets in the process of atherosclerosis, and has important clinical value to delay the atherosclerotic thrombosis.     

Effects of sodium metabisulfite on sodium current in acutely isolated rat hippocampal CA1 neurons

AIM: To study the effects of sodium metabisulfite (SMB), sulfur dioxide (SO2) and its derivatives in vivo, sodium bisulfite and sulfite, on Na+ currents. The effects of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) against SMB were also studied in freshly dissociated hippocampal CA1 neurons in rats. METHODS: The whole-cell patch-clamp techniques were used in the experiments. RESULTS: ① SMB increased the voltage-activated Na+ currents in a concentration-and voltage-dependent manner. The amplitudes of Na+ currents was increased (22.36±3.28) % and (65.05±5.75)% (n=10) by SMB at 2 μmol/L and 20 μmol/L, respectively. ② SMB (10 μmol/L) did not affect the activation process, but changed the inactivation process significantly. Before and after application of 10 μmol/L SMB, the half-inactivation voltage was (-82.38±0.54) mV and (-69.39±0.41) mV (n=10, P<0.01). However, the slope factor remained unchanged. ③ SOD, CAT and GPx partly inhibited the incremental effect of SMB on Na+ currents. CONCLUSION: SMB increases Na+ currents and inhibits the steady-state inactivation, which contributes to the increase in the excitibility of neuronal cells. Its mechanism may involve oxidative damage caused by sulfur-and oxygen-centered free radicals in the rat hippocampal CA1 neurons.     

Effects of Chlamydia pneumoniae infection and hyperlipidemia on the expression of NF-κB and AP-1 in myocardium

AIM: To investigate the effects of Chlamydia pneumoniae infection and hyperlipidemia on the expression of NF-κB and AP-1 in the myocardium. METHODS: The indirect immunofluorescence method was used to examine wild C57BL/6J mice infected with Chlamydia pneumoniae and fed with an atherogenic diet. The expression of the subunit of NF-κB, P50, and c-Fos in the murine myocardium was observed. RESULTS: Chlamydia pneumoniae infection and hyperlipidemia induced the activation of NF-κB and AP-1 in murine myocardium. P50 and c-Fos were not detected in the controls, but there were different levels of positive expression in the experiments (P<0.01, compared to the controls). No statistical significance among the experiments was observed. CONCLUSION: In early time of Chlamydia pneumoniae infection and hyperlipidemia, the inflammatory pathway is already activated in the myocardium.