OBJECTIVETo explore the synergistic protection of Danhong Injection (丹红注射液, DHI) and ischemic postconditioning on myocardial reperfusion injury in minipigs.

METHODSAcute myocardial infarction model was made by balloon occlusion in left anterior descending coronary artery (LAD) of minipigs, and then postconditioning was simulated through inflation/deflation of the angioplasty balloon. Minipigs were divided into four groups: the sham operation group (SH group), the ischemia/reperfusion group (I/R group), the ischemic postconditioning group (POC group) and DHI combined with ischemic postconditioning group (PAD group, DHI 20 mL through ear vein), six in each group. After 24-h continuous observation, myocardial infarction size was assessed by triphenyltetrazolium staining (TTC). Morphological changes of ischemic myocardium were observed by light microscopy, and cardiomyocyte ultrastructure was studied with electron microscopy. The superoxide dismutase (SOD) and malondialdehyde (MDA) activity in heart homogenates were measured by a biochemical method.

RESULTSThe myocardial infarction size was smaller in the POC group than in the I/R group (0.26 ± 0.02 vs. 0.37 ± 0.09, P<0.05), and the PAD group (0.14 ± 0.08) displayed a significantly reduced infarction size relative to the I/R group (P<0.01) and POC group (P<0.05). The damage of myocardial tissue was severe in the I/R group shown by light and electron microscopy: myocardial fibers disorder, sarcoplasmic dissolution, myofilament fracture, mitochondria swelling and even vacuolization formation and a large number of inflammatory cell infiltrations. Compared with the I/R group, reduction of reperfusion injury in the PAD group included more orderly arranged myocardial fibers, less infiltration of inflammatory cells and maintenance of mitochondrial integrity. Compared with the I/R group, the damage of myocardial tissue in the POC group was improved, but not as significant as that in the PAD group. SOD levels in the POC group and the PAD group were significantly higher than those in the I/R group (96.96 ± 13.43, 112.25 ± 22.75 vs. 76.32 ± 10.63, P<0.05), and MDA was significantly lower in the POC group and the PAD group compared to the I/R group (1.27 ± 0.19, 1.09 ± 0.21 vs. 1.47 ± 0.16, P<0.05).

CONCLUSIONDHI and ischemic postconditioning show a synergistic cardioprotection on myocardial reperfusion injury in minipigs.

Animals ; Coronary Angiography ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Injections ; Ischemic Postconditioning ; Malondialdehyde ; metabolism ; Myocardial Infarction ; complications ; pathology ; Myocardial Reperfusion Injury ; complications ; drug therapy ; prevention & control ; Myocardium ; enzymology ; pathology ; ultrastructure ; Superoxide Dismutase ; metabolism ; Swine ; Swine, Miniature

OBJECTIVETo evaluate the anti-apoptotic effect of aldehyde dehydrogenase 2 (ALDH2) on myocardial ischemia/reperfusion (I/R) injury in diabetic rats.

METHODSNormal male SD rats were divided into normal, diabetes and ethanol (the agonist of ALDH2) + diabetes groups. In the latter two groups, diabetes was induced by an intraperitoneal injection of 55 mg/kg STZ. Four weeks after the modeling, myocardial I/R was mimicked ex vivo, and lactate dehydrogenase (LDH) content in the coronary flow was determined. The activities of caspase-3 and ALDH2 were evaluated, and the expressions of Bcl-2 and Bax mRNA in the left anterior myocardium were detected using RT-PCR.

RESULTSIn diabetic group, LDH release and caspase-3 activity were increased, while ALDH2 activity and Bcl-2/Bax mRNA expression were decreased as compared to those in normal control group. Compared with the diabetic group, ALDH2 agonist ethanol significantly reduced LDH release and caspase-3 activity, increased ALDH2 activity and Bcl-2/Bax mRNA expression.

CONCLUSIONIn diabetic rats, enhanced ALDH2 expression can offer mycardial protection possibly in relation to suppress cell apoptosis.

Aldehyde Dehydrogenase ; metabolism ; Aldehyde Dehydrogenase, Mitochondrial ; Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Diabetes Mellitus, Experimental ; complications ; enzymology ; Ethanol ; pharmacology ; Male ; Mitochondrial Proteins ; agonists ; metabolism ; Myocardial Ischemia ; enzymology ; etiology ; Myocardial Reperfusion Injury ; enzymology ; pathology ; prevention & control ; Myocardium ; enzymology ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

Intestinal ischemia-reperfusion (I/R) is an important event in the pathogenesis of multiple organ dysfunction syndrome (MODS). The aim of this study is to determine the effects of ginsenoside Rb1 on liver injury induced by intestinal I/R in rats. Adult male Wistar rats were randomly divided into four groups: (1) a control, sham-operated group (sham group); (2) an intestinal I/R group subjected to 1 h intestinal ischemia and 2 h reperfusion (I/R group); (3) a group treated with 20 mg/kg ginsenoside Rb1 before reperfusion (Rb1-20 group); and (4) a group treated with 40 mg/kg ginsenoside Rb1 before reperfusion (Rb1-40 group). Liver and intestinal histology was observed. Aspartate aminotransferase (AST), alanine aminotransferase (ALT) level in serum and malondialdehyde (MDA) level in intestinal tissues were measured. Myeloperoxidase (MPO), TNF-alpha, MDA level and immunohistochemical expression of NF-kappa B and intracellular adhesion molecale-1 (ICAM-1) in liver tissues was assayed. In addition, a western blot analysis of liver NF-kappa B expression was performed. Results indicated intestinal I/R induced intestinal and liver injury, which was characterized by increase of AST and ALT in serum, MDA level in intestine, MPO, TNF-alpha and MDA level and ICAM-1 and NF-kappa B expression in the liver tissues. Ginsenoside Rb1 (20, 40 mg/kg) ameliorated liver injury, decreased MPO, TNF-alpha and MDA level, NF-kappa B and ICAM-1 expression in liver tissues. In conclusion, ginsenoside Rb1 ablated liver injury induced by intestinal I/R by inhibiting NF-kappaB activation.
Alanine Transaminase/blood ; Animals ; Aspartate Aminotransferases/blood ; Biological Markers/metabolism ; Ginsenosides/*pharmacology ; Intercellular Adhesion Molecule-1/metabolism ; Intestines/*blood supply/metabolism/pathology ; Ischemia/metabolism/pathology ; Liver/enzymology/pathology ; Liver Diseases/etiology/*pathology ; Male ; Malondialdehyde/metabolism ; NF-kappa B/*antagonists & inhibitors/metabolism ; Peroxidase/metabolism ; Rats ; Rats, Wistar ; Reperfusion Injury/complications/*pathology ; Tumor Necrosis Factor-alpha/metabolism

AIMTo study the function of c-Jun N-terminal kinase 3 (JNK3) in the process of ischemic/reperfused heart injury and the mechanism underlying the protective action of magnesium lithospermate B (MTB), a bioactive compound isolated from Danshen.

METHODSBy in situ hybridization, JNK3 mRNA was detected in the ventricular preparations of the Langendorff ischemic/reperfused rat heart. The inhibitory effect of MTB on the expression of JNK3 mRNA was also investigated.

RESULTSThe purple and blue hybridization signals were located in the cytoplasm of the cardiomyocytes, which were weaker in the non-perfused hearts and stronger in the hearts encountered 30 min of ischemia and 30 min of reperfusion. Image analysis showed that the expression of JNK3 mRNA in the cardiomyocytes increased after 30 min of ischemia and 30 min of reperfusion, which showed significant difference compared with that in the cardiomyocytes of the non-perfused heart and the control heart (P < 0.05). Treatment with of 0.1, 1 and 10 mumol.L-1 MTB abolished the elevation of JNK3 mRNA expression in the ischemic/reperfused heart (P < 0.05).

CONCLUSIONJNK3 may be another component in the signal transduction pathway of ischemia/reperfusion induced cardiomyocyte apoptosis. MTB may protect the heart from ischemia/reperfusion injury by reducing apoptosis through inhibition of the JNK3 activity.

Animals ; Cardiotonic Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; Male ; Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; genetics ; metabolism ; Myocardial Ischemia ; complications ; Myocardial Reperfusion Injury ; enzymology ; etiology ; pathology ; Myocytes, Cardiac ; drug effects ; enzymology ; RNA, Messenger ; biosynthesis ; drug effects ; Rats ; Rats, Sprague-Dawley

BACKGROUND/AIMS: Ischemic preconditioning (IP) decreases severity of liver necrosis and has anti-apoptotic effects in previous studies using liver regeneration in normal rats. This study assessed the effect of IP on liver regeneration after hepatic resection in cirrhotic rats. METHODS: To induce liver cirrhosis, thioacetamide (300 mg/kg) was injected intraperitoneally into Sprague-Dawley rats twice per week for 16 weeks. Animals were divided into four groups: non-clamping (NC), total clamping (TC), IP, and intermittent clamping (IC). Ischemic injury was induced by clamping the left portal pedicle including the portal vein and hepatic artery. Liver enzymes alanine transaminase (ALT) and aspartate aminotransferase (AST) were measured to assess liver damage. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining for apoptosis and proliferating cell nuclear antigen (PCNA) staining for cell replication were also performed. RESULTS: Day-1 ALT and AST were highest in IP, however, levels in NC and IC were comparably low on days 1-7. There was no significant correlation of AST or ALT with experimental groups (P=0.615 and P=0.186). On TUNEL, numbers of apoptotic cells at 100x magnification (cells/field) were 31.8+/-24.2 in NC, 69.0+/-72.3 in TC, 80.2+/-63.1 in IP, and 21.2+/-20.8 in IC (P<0.05). When regeneration capacity was assessed by PCNA staining, PCNA-positive cells (cells/field) at 400x were 3.4+/-6.0 in NC, 16.9+/-69 in TC, 17.0+/-7.8 in IP and 7.4+/-7.6 in IC (P<0.05). CONCLUSIONS: Although regeneration capacity in IP is higher than IC, the liver is vulnerable to ischemic damage in cirrhotic rats. Careful consideration is needed in applying IP in the clinical setting.
Alanine Transaminase/blood ; Animals ; Apoptosis ; Aspartate Aminotransferases/blood ; Constriction ; Hepatectomy/methods ; Hepatic Artery ; *Ischemic Preconditioning ; Liver/blood supply ; Liver Cirrhosis, Experimental/chemically induced/complications/*pathology ; *Liver Regeneration ; Proliferating Cell Nuclear Antigen/metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury/complications/enzymology/pathology ; Thioacetamide/toxicity

AIMTo explore the protective effect of propyl gallate against neuronal injury in the boundary zone of the infarction area in the rat cerebral ischemia-reperfusion model and its possible mechanism.

METHODSTransient focal ischemia induced by middle cerebral artery occlusion in the rats was established by ligation of the left internal carotid artery for 2 h. Rats were treated by propyl gallate with different doses (23.5, 47 and 94 micromol x kg(-1)) for three days before operation. Coronal brain sections were collected after 1 , 2, 4, 6, 12 and 24 h of reperfusion, neuronal injury in the boundary zone of the infarction area was evaluated by TUNEL and Nissl staining. The expression of activated Caspase-3, total SAPK/JNK, p38MAPK and their phosphorylation (Thr183/Tyr185, Thr180/Tyr182) was investigated by immunohistochemistry and Western blotting with corresponding antibodies.

RESULTSAlthough SAPK/JNK immunoreactivity did not increase at each time point in the boundary zone of the infarction area after reperfusion, p-SAPK/JNK immunoreactivity increased significantly at 1 h and then decreased gradually, and p38MAPK immunoreactivity was enhanced at each time point, peaked at 6 h. Expression of p-p38MAPK peaked at 6 h. Activated Caspase-3 immunoreactivity appeared at 6 h in the boundary zone of the infarction area and peaked at 12 h. TUNEL positive neurons were observed at 12 h and became more abundant at 24 h. The number of Nissl positive neurons decreased gradually and apoptosis ratio of neurons peaked at 24 h. Propyl gallate reduced the immunoreactivity of SAPK/JNK, p-SAPK/JNK, p38MAPK and p-p38MAPK markedly at 1 and 6 h. Propyl gallate with doses of 47 and 94 micromol x kg(-1) were more effective.

CONCLUSIONInhibition on the activation of SAPK/JNK and p38MAPK is the possible protective mechanism of propyl gallate against neuronal injury induced by cerebral ischemia-reperfusion.

Animals ; Apoptosis ; drug effects ; Brain Ischemia ; etiology ; Caspase 3 ; metabolism ; Enzyme Activation ; drug effects ; Infarction, Middle Cerebral Artery ; complications ; MAP Kinase Kinase 4 ; metabolism ; Male ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Propyl Gallate ; pharmacology ; Putamen ; pathology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; enzymology ; etiology ; pathology ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo observe the effect of ligustrazine on cell proliferation in the subventricular zone (SVZ) and dentate gyrus (DG) and nNOS expression in rat brain after cerebral ischemia-reperfusion injury.

METHODSMale SD rats were randomly divided into normal control group, sham operation group, model group and ligustrazine treatment group. The latter two groups were further divided into 5 subgroups for observation at 1, 3, 7, 14 and 21 days after reperfusion following a 2-hour middle cerebral artery occlusion (MCAO). The cells in S phase were labeled with BrdU, and immunohistochemistry was employed to detect BrdU- and nNOS-positive cells. The numbers of BrdU-positive cells in the SVZ and DG were measured. The expression of nNOS was detected by Western blotting.

RESULTSnNOS expression increased significantly in the model group as compared to the sham operation group (P<0.05), and ligustrazine treatment significantly lowered the expression level in comparison with the model group (P<0.05). Compared with the model group, a significant increase in BrdU-positive cells occurred in the SVZ of rats 1 and 3 days after igustrazine treatment (P<0.05), along with an increase of DG BrdU-positive cells.

CONCLUSIONLigustrazine significantly restrains ischemia-reperfusion injury-induced nNOS activity enhancement and promotes cell proliferation in the SVZ and DG of adult rats after ischemia-reperfusion injury.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Blotting, Western ; Brain ; blood supply ; drug effects ; enzymology ; Brain Ischemia ; complications ; Cell Proliferation ; drug effects ; Cerebral Ventricles ; blood supply ; drug effects ; pathology ; Dentate Gyrus ; blood supply ; drug effects ; pathology ; Immunohistochemistry ; Male ; Nerve Regeneration ; drug effects ; Nitric Oxide Synthase Type I ; biosynthesis ; Pyrazines ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; physiopathology