This study was to investigate the effect of peoniflorin on the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream signal molecules in the hippocampus of Alzheimer's disease (AD) rats for exploring the mechanism of peoniflorin protecting hippocampal neurons. AD model rats were established by bilateral intrahippocampal injection of beta-amyloid(1-42) (Abeta(1-42)) and divided randomly into 3 groups: AD model group, peoniflorin low-dose (15 mg x kg(-1)) group and peoniflorin high-dose (30 mg x kg(-1)) group. The vehicle control rats were given bilateral intrahippocampal injection of solvent with the same volume. After peoniflorin or saline was administered (ip) once daily for 14 days, the hippocampuses of all animals were taken out for measuring the expressions of Nrf2, heme oxygenase-1 (HO-1) and gamma-glutamylcysteine synthethase (gamma-GCS) mRNA by reverse transcription PCR, determining the contents of glutathione (GSH), malondialdehyde (MDA) and carbonyl protein (CP) using colorimetric method, and for assaying the expressions of neuronal apoptosis inhibitory protein (NAIP) and Caspase-3 by immunohistochemical staining method. The results showed that peoniflorin markedly increased the expressions of Nrf2, HO-1 and gamma-GCS mRNA, enhanced the level of GSH and decreased the contents of MDA and CP in the hippocampus, as compared with the model group. Peoniflorin also improved the NAIP expression and reduced the Caspase-3 expression in the hippocampus neurons. In conclusion, peoniflorin protects against the Abeta(1-42)-mediated oxidative stress and hippocampal neuron injury in AD rats by activating the Nrf2/ARE pathway.
Alzheimer Disease ; chemically induced ; metabolism ; physiopathology ; Amyloid beta-Peptides ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Caspase 3 ; metabolism ; Glucosides ; pharmacology ; Glutamate-Cysteine Ligase ; genetics ; metabolism ; Glutathione ; metabolism ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; Hippocampus ; metabolism ; Male ; Malondialdehyde ; metabolism ; Monoterpenes ; pharmacology ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Neuronal Apoptosis-Inhibitory Protein ; metabolism ; Neurons ; metabolism ; Oxidative Stress ; drug effects ; Peptide Fragments ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

BACKGROUND/AIMS: Renal hypoxia is involved in the pathogenesis of diabetic nephropathy. Pentoxifyllin (PTX), a nonselective phosphodiesterase inhibitor, is used to attenuate peripheral vascular diseases. To determine whether PTX can improve renal hypoxia, we investigated its effect in the streptozocin (STZ)-induced diabetic kidney. METHODS: PTX (40 mg/kg, PO) was administered to STZ-induced diabetic rats for 8 weeks. To determine tissue hypoxia, we examined hypoxic inducible factor-1alpha (HIF-1alpha), heme oxygenase-1 (HO-1), vascular endothelial growth factor (VEGF), and glucose transporter-1 (GLUT-1) levels. We also tested the effect of PTX on HIF-1alpha in renal tubule cells. RESULTS: PTX reduced the increased protein creatinine ratio in diabetic rats at 8 weeks. HIF-1alpha, VEGF, and GLUT-1 mRNA expression increased significantly, and the expression of HO-1 also tended to increase in diabetic rats. PTX significantly decreased mRNA expression of HIF-1alpha and VEGF at 4 and 8 weeks, and decreased HO-1 and GLUT-1 at 4 weeks. The expression of HIF-1alpha protein was significantly increased at 4 and 8 weeks in tubules in the diabetic rat kidney. PTX tended to decrease HIF-1alpha protein expression at 8 weeks. To examine whether PTX had a direct effect on renal tubules, normal rat kidney cells were stimulated with CoCl2 (100 microM), which enhanced HIF-1alpha mRNA and protein levels under low glucose conditions (5.5 mM). Their expressions were similar even after high glucose (30 mM) treatment. PTX had no effect on HIF-1alpha expression. CONCLUSIONS: PTX attenuates tubular hypoxia in the diabetic kidney.
Animals ; Anoxia/*drug therapy/enzymology/etiology/genetics ; Cell Line ; Cobalt/pharmacology ; Diabetes Mellitus, Experimental/*complications ; Diabetic Nephropathies/*drug therapy/enzymology/etiology/genetics ; Disease Models, Animal ; Gene Expression Regulation/drug effects ; Glucose/metabolism ; Glucose Transporter Type 1/genetics ; Heme Oxygenase (Decyclizing)/genetics/metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics/metabolism ; Kidney Tubules/*drug effects/enzymology ; Male ; Pentoxifylline/*pharmacology ; Phosphodiesterase Inhibitors/*pharmacology ; RNA, Messenger/metabolism ; Rats ; Rats, Sprague-Dawley ; Streptozocin ; Time Factors ; Vascular Endothelial Growth Factor A/genetics

Animals ; Animals, Newborn ; Cells, Cultured ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; Ischemic Preconditioning, Myocardial ; methods ; Myocardial Ischemia ; physiopathology ; Myocardial Reperfusion Injury ; prevention & control ; Myocytes, Cardiac ; cytology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar

AIMTo investigate the possible role of rate-limiting enzyme of heme metabolism and globin in the development of the low hemoglobin (Hb), red blood (cell) count (RBC) and hematocrit (Hct) after long-term exercise, and effect of nutrition supplement on sports anemia.

METHODSMale Wistar rats were randomly assigned to three groups (n = 10): control (C), exercise (P) and exercise + nutrition (G). Animals in the P and G groups started treadmill running at 30 m/min, 0% grade, 1 min/time. Running time was gradually increased with 2 min/time during initial 5 weeks and final 4 weeks. In addition, running frequency was 2 times/day except initial 2 weeks. At the end of eleventh week, gene expression of 5-aminolevulinate synthase (ALAS), ferrochelatase, alpha-globin and beta-globin in bone marrow were measured with RT-PCR. Mean-while heme oxygenase 1 (HO-1) activity in liver was measured with immunohistochemical method.

RESULTSEleven weeks of exercise induced a significant increase in HO-1 and a significant increase in gene expression of beta-globin (P < 0.01, P < 0.05, respectively). Treatment with anti-sports anemia compound dosage led to no significant differences in rate-limiting enzyme of heme metabolism and globin in the exercised rats. The G group had a significantly higher HO-1 level in liver than the C group (P < 0.01). These finds showed that exercise was associated with no significant difference in heme synthetase and alpha-globin gene expression, and significant difference in heme catabolic enzyme and beta-globin gene expression.

CONCLUSIONThe increase of HO-1 activity in liver might be one of the causes of the lower Hb, RBC and Hct status in exercised rats.

5-Aminolevulinate Synthetase ; genetics ; metabolism ; Anemia ; etiology ; metabolism ; physiopathology ; Animals ; Dietary Supplements ; Ferrochelatase ; genetics ; metabolism ; Gene Expression Regulation, Enzymologic ; physiology ; Globins ; metabolism ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; Hydroxymethylbilane Synthase ; genetics ; metabolism ; Male ; Motor Activity ; Physical Conditioning, Animal ; adverse effects ; Random Allocation ; Rats ; Rats, Wistar

AIMTo investigate the changes of lipid peroxidation level and expression of heme oxygenase-1 of the rat liver with chronic hypoxia and hypercapnia, and the effects of Safflower injection (a compond of Chinese Traditional medicine).

METHODSThirty male SD rats weighing 180 approximately 220 g were divided into three groups (n=10): control group (N group), chronic hypoxia and hypercapnia for four weeks group(F group), and Safflower injection group (H group). SOD and MDA in liver tissue were measured by spectrophotometric method. And methods Immunohistochemical assay was used to detect the distribution of HO-1 protein. Pathological changes in liver tissues were observed in HE staining section. The mRNA expressions of HO-1 in liver were detected by semi-quantitative RT-PCR.

RESULTSThe activity of SOD of the liver in F group were significantly lower than those in N group, and the content of MDA were significantly higher. The activity of SOD of the liver in H group were significantly higher than those in F group, and the content of MDA were significantly lower. In F group there were multiple dispersed immunoreactivity cells in liver. And compared to those in F group, the immunoreactivity cells were significantly decreased in H group. HE staining revealed that there were many hepatocytes with obvious adipose degeneration. Hepatic pathological damage in H group was slighter than that in F group. The expression of HO-1 mRNA of the liver in F group were significantly higher than those in N group (P < 0.01), and those in H group were significantly lower than those in F group (P < 0.01) .

CONCLUSIONChronic hypoxia and hypercapnia increases the level of oxidative stress. Safflower injection have a protective effect, maybe because of the accommodation of the expression of HO-1 of the liver and the elimination of free radicals.

Animals ; Carthamus tinctorius ; chemistry ; Chronic Disease ; Drugs, Chinese Herbal ; pharmacology ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; Hypercapnia ; physiopathology ; Hypoxia ; physiopathology ; Lipid Peroxidation ; drug effects ; Liver ; metabolism ; pathology ; Male ; Oxidative Stress ; physiology ; Protective Agents ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

Free radical hypothesis of aging emphasized that the age-related accumulation of free radicals results in cell injury. Alzheimer's disease (AD) is the most common form of neurodegenerative disease characterized by impaired cognition and memory of the elderly. Aging is a key risk factor in AD. Substantial evidence suggests that imbalance between free radical formation and clearance promotes AD pathogenesis. The brain overcomes oxidative stress by inducing expression of a set of genes called vitagenes. The protein products of vitagenes include heat shock proteins, heme oxygenases and thioredoxin systems, which serve as endogenous lifeguard of cells. This paper is a review of the expression and function of vitagenes in aging and AD brain, as well as relevant pharmacological study.
Aging ; genetics ; metabolism ; Alzheimer Disease ; genetics ; metabolism ; Brain ; metabolism ; Heat-Shock Proteins ; genetics ; metabolism ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; Humans ; Oxidative Stress ; Thioredoxins ; genetics ; metabolism

OBJECTIVETo study the mechanism of paraquat-induced renal injury in rats.

METHODSAdult healthy Sprague-Dawley (SD) rats (female and male in half) were randomly divided into two groups, the control group and the paraquat poisoned group. The rats in the paraquat poisoned group were treated with PQ (25 mg/kg) intraperitoneally while the rats in the control group were treated with the same dose of normal saline. Its histopathological change was observed and the expression of HO-1 and the mRNA expression of HO-1 were detected by RT-PCR at 3rd h, 6th h, 12th h, on 1st d, 2nd d, 3rd d and 5th d.

RESULTS(1) In the control group, the tissue structure was clear without edema, vacuolar degeneration, cloudy swelling and necrosis. In the paraquat poisoned group, there were obvious lesions in the renal tubule of cortical part, including cellular swelling, the narrow cannula, the mesenchymal congestion and edema. These pathologic changes gradually became more severe, reached the peak on the 1st day, and did not relieve until the end of this study; there was the karyopyknosis and the cyto-architecture disappeared in some severe cases; Some glomerulus and medulla were also involved. (2) In the control group, there was no or weak expression of HO-1 and HO-1 mRNA. At the 3rd hour, the expressions of HO-1 in the paraquat poisoned group were observed in the membrane and cytoplasm of renal tubular epithelial cell of cortical part. Immunohistochemistry score (IHS) in the paraquat poisoned group was higher than that in the control group (P<0.05), except the HIS of the 5th day. At the 3rd hour, the expression of HO-1 mRNA increased, reached the peak on the 1st day, and then decreased. The expression of HO-1 mRNA was (0.53 +/- 0.21), (0.55 +/- 0.31), (0.56 +/- 0.22), (0.64 +/- 0.14) and (0.43 +/- 0.25) at the time point other than on the 3rd and 5th day. It showed statistical difference between the paraquat poisoned group and the control group from the 3rd hour to the 2nd day (P<0.05).

CONCLUSIONThe mechanism of paraquat induced-renal injury is multiple. The higher expression of HO-1 and HO-1 mRNA were involved in the procedures of paraquat-induced renal injury.

Animals ; Female ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; Kidney ; enzymology ; pathology ; Male ; Paraquat ; poisoning ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the possible protective effect and mechanism of ginsenoside Rb1 against oxidative damage and renal interstitial fibrosis on rats with unilateral ureteral obstruction (UUO).

METHODSIn total, 80 male rats were randomly divided into 4 groups, 20 in each group: the sham operated group (SOR), UUO group, UUO with ginsenoside Rb1 treatment group (treated with intraperitoneal injection of 50 mg/ kg daily) and UUO with Losartan treatment group (as the positive control, treated with 20 mg/kg by gastrogavage per day). The rats were randomly sacrificed on day 3, 7 and 14 after surgery, respectively. The histopathologic changes of renal interstitial tissues were observed with Masson staining. The mRNA of transforming growth factor beta 1 (TGF-beta 1), collagen I and fibronectin were reversed transcribed and quantified by Real-time PCR. Enzyme-linked immunosorbent assay was used to quantitatively detect TGF-beta 1 and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels. P47phox protein expression was assessed by immunohistochemistry and Western blot analysis.

RESULTSIn the UUO model, the obstructed kidney showed typical features of progressive renal tubulointerstitial fibrosis, and the levels of TGF-beta1, collagen I and fibronectin increased (P<0.05). As compared with the UUO group, ginsennoside Rb1 significantly inhibited the interstitial fibrosis including tubular injury and collagen deposition, and decreased the levels of TGF-beta1 (P<0.05). Ginsenoside Rb1 also inhibited the heme oxygenase (HO-1) and 8-OHdG, two markers of oxidative stress (P<0.05). Moreover, ginsenoside Rb1 suppressed the expression of p47phox, a subunit of nicotinamide adeninedinucleotide phosphate (NADPH) oxidase (P<0.05).

CONCLUSIONGinsenoside Rb1 can obviously inhibit renal interstitial fibrosis in rats with UUO, its mechanism possibly via against the oxidative damage and suppressing TGF-beta1 expression.

Animals ; Deoxyguanosine ; analogs & derivatives ; urine ; Drug Evaluation, Preclinical ; Fibrosis ; genetics ; metabolism ; prevention & control ; Gene Expression Regulation ; drug effects ; Ginsenosides ; therapeutic use ; Heme Oxygenase (Decyclizing) ; metabolism ; Kidney ; drug effects ; metabolism ; pathology ; Kidney Diseases ; etiology ; genetics ; pathology ; prevention & control ; Male ; Models, Biological ; NADPH Oxidases ; genetics ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Saponins ; therapeutic use ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Ureteral Obstruction ; complications ; drug therapy ; genetics ; metabolism

OBJECTIVETo cultivate human umbilical vein endothelial cells (HUVECs) in the serum of overfatigue rats with the intervention of Tongxinluo superfine powder (TXLSP). By examining the variation of the activity of JNK/c-Jun/HO-1 pathway, the possible mechanisms of vascular endothelial dysfunction under overfatigue conditions and the intervening effect of TXLSP were explored.

METHODSThe HUVECs were randomly divided into the normal control group, the model group, the SP600125 (a specific antagonist of JNK) group, the TXLSP group and the TXLSP + SP600125 group. The content of carboyhemoglobin (COHb) and the leak rate of lactic dehydrogenase (LDH) in different groups were measured. The mRNA and protein expression of JNK, c-Jun, HO-1 and the phosphorylation level of c-Jun (P-c-Jun) were detected using Western blot and PCR methods.

RESULTSCompared with the normal control group, the COHb level in supernatant was increased significantly in the model group, and the expression of HO-1, JNK, c-Jun mRNA and corresponding proteins and P-c-Jun were also increased remarkably. The increases in these parameters were significantly decreased by SP600125. TXLSP showed remarkable up-regulation on the expression of JNK, c-Jun, P-c-Jun and HO-1 mRNA and their protein expression. Compared with the SP600125 group, the expressions of JNK, c-Jun, P-c-Jun and HO-1 mRNA and its protein in the TXLSP+SP600125 group were significantly increased at different time points (P<0.05, P<0.01).

CONCLUSIONSThe vascular endothelial dysfunction under overfatigue conditions is related to the activity of the JNK/c-Jun/HO-1 pathway. One of the mechanisms of TXLSP in improving the vascular endothelial function is to adjust the activity of the JNK/c-Jun/HO-1 pathway at gene and protein levels.

Animals ; Blood Proteins ; pharmacology ; Cells, Cultured ; Cytoprotection ; drug effects ; genetics ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Endothelial Cells ; drug effects ; metabolism ; Fatigue ; blood ; metabolism ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; physiology ; Humans ; JNK Mitogen-Activated Protein Kinases ; genetics ; metabolism ; physiology ; Male ; Particle Size ; Powders ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins c-jun ; genetics ; metabolism ; physiology ; Rats ; Rats, Wistar ; Serum ; metabolism ; physiology ; Signal Transduction ; drug effects ; genetics ; physiology ; Umbilical Veins ; cytology ; drug effects ; metabolism

OBJECTIVETo study the expression of heme oxygenase (HO) in the corpus cavernosum of castrated rats and to investigate its role in androgen deficiency-mediated erectile dysfunction.

METHODSForty 10-week old SD rats were randomly divided into Groups A (2-week sham operation), B (4-week sham operation), C (2-week castrated) and D (4-week castrated). The serum testosterone level was determined, and the expressions of HO and nNOS in the corpus cavernosum of the castrated rats were detected by immunohistochemical staining and RT-PCR 2 and 4 weeks after the operation.

RESULTSCompared with the sham operation groups, the castrated rats showed a significant reduction in the level of serum testosterone, (A vs. C: [283.222 +/- 117.171] ng/dl vs. [7.117 +/- 3.700] ng/ dl; B vs. D: [289.280 +/- 87.413] ng/dl vs [48.826 +/- 19.477] ng/dl) (P < 0.01), the expressions of HO-1 and HO-2 proteins (P < 0.01) and the expressions of HO-1, HO-2 and nNOS mRNA (P < 0.01).

CONCLUSIONAndrogen can partly regulate the erectile function of the penis via the HO-CO system.

Animals ; Heme Oxygenase (Decyclizing) ; genetics ; metabolism ; Male ; Nitric Oxide Synthase Type I ; metabolism ; Orchiectomy ; Penis ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; blood