Expression of MMP—2,TIMP—2 protein and the ratio of MMP—2／TIMP—2 in gallbladder carcinoma and their significance

AIM: To study the correlation between expression of MMP-2, TIMP-2 protein and the ratio of MMP-2/TIMP-2 and clinicalpathological parameters of patients with gallbladder carcinoma.METHODS: Carcinomas (n=45) and polypoid lesions (n=15) of the gallbladder were studied for the expression of MMP-2 and TIMP-2 protein by immunohistochemical avidin-biotin-complex method and image analysis. Clinicalpathological data of patients with gallbladder carcinoma such as histological type, grade of differentiation, level of infiltration, liver invasion and lymph node involvement, etc, were recorded.RESULTS: There was significant difference between the average level (1.123±0.108 VS 1.030±0.054, P=0.002) of MMP-2, the ratio (1.050±0.013 VS0.937±0.078, P=0.003) of MMP-2/TIMP-2 in gallbladder carcinomas and in polypoid lesions of the gallbladder. Significant difference was found between the expression of MMP-2 in early stage and advanced tumors, but there was no correlation between MMP-2 protein expression and histological type, differentiation degree, infiltration level, lymph node involvement or liver invasion. Although no difference was observed between TIMP-2 expression and histological type or differentiation degree, signific ant difference was found between TIMP-2 expression and different Nevin stage, infiltration level, local lymph node involvement or liver invasion (1.168±0.067 VS1.048±0.075, 1.170±0.062 vs 1.039±0.06g, 1.039±0.076 VS1.147±0.083, 1.048±0.074 vs 1.103±0.095, P＜0.05). MMP-2/TIMP-2 ratio did not correlate with histological type, grade of differentiation and liver invasion, but significant differences were found between MMP-2/TIMP-2 ratio and different Nevin stage, infiltration level and lymph node involvement in patients with carcinoma of gallbladder.CONCLUSION: TIMP-2 and MMP-2/TIMP-2 ratio could reflect more accurately biological characteristic of gallbladder carcinoma and MMP-2/TIMP-2 ratio might be a new significant marker in early diagnosis, in the judgment of invasion or metastasis and the estimate of prognosis in patients with gallbladder carcinomas.     

The role of endotoxin,TNF—α,and IL—6 in inducing the state of growth hormone insensitivity

AIM：Critical illnesses such as sepsis,trauma,and burns cause a growth hormone insensitivity,wehich leads to an increased negative nitrogen balance.Endotoxin is generously released into blood under these conditions and stimulates the production of proinflammatory cytokines such as TNF-α,IL-6,and IL-1 which may play a very important role in inducing the growth hormone insensitivity,The objective of this current study was to investigate the role of endotoxin,TNFα and IL-6 in inducing the growth hormone insensityvity at the receptor and post-receptor levels.METHODS:Spageu-Dawley rats were injected with endotoxin,TNF-α,and IL-6,respectively and part of rats injected with endotoxin was treated with exogenous somatotropin simultaneously,All rats were killed at different time points,The expression of IGF-I,GHR,SOCS-3 and β-actin mRNA in the liver was detected by RT-PCR and the GH levels were measured by radioimmunoassay,the levels of TNF-α and IL-6 were detected by ELISA.RESULTS:There was no significant difference in serous GH levels between experimental group and control rats after endotoxin injection,However,liver IGF-1 mRNA expression had been obviously down-regulated in endotoxeminc rats.Liver GHR mRNA expression also had a predominant downregulation after endotoxin injection,The lowest regulation of liver IGF-I mRNA expression occurred at 12h after LPS injection,being decreased by 53% compared with control rats.For GHR mRNA expression,the lowest expression occurred at 8h and had a 81% decrease.Although SOCS-3 mRNA was weakly expressed in control rats,it was strongly up-regulated after LPS injection and had a 7.84 times increase compared with control rats.Exogenous GH could enhance IGF-1 mRNA expression in control rats,but if did fail to prevent the decline in IGF-1 mRNA expression in endotoxemic rats,Endotoxin stimulated the production of TNF-α and IL-6,and the elevated IL-6 levels was shown a positive correlation with increased SOCS-3 mRNA expression.The liver GHR mRNA expression was obviously down-regulated after TNF-α iv injection and had a 40 decresase at 8 h,but the liver socs-3 mRNA expression was the 4.94 times up-regulation occurred at 40 min after IL-6 injection.CONCLUSION:The growth homone insenstivity could be induced by LPS injection,which was associated with down regulated GHR mRNA expression at receptor level and with up-regulated SOCS-3 mRNA expression at post-receptor level The in vivo biological activities of LPS were mediated by TNF-α and IL-6 indirectly,and TNF-αand IL-6 may exert their effects on.the receptor and post-receptor levels respectively.     

Effect of apoptosis on gastric adenocarcinoma cell line SGC—7901 induced by cis—9,trans—11—conjugated linoleic acid

AIM: To determine the effect of apoptosis on gastric cancer cells (SGC-7901) induced by cis-9, trans-11-conjugated linoleic acid (c9, t11-CLA) and its possible mechanism in the inhibition of cancer cells growth. METHODS: Using cell culture, flow cytometery and immunocytochemical techniques, we examined the cell growth, frequency of apoptosis and distribution of cell cycle, expression of ki67, bcl-2, Fas, and c-myc of SGC-7901 cells which were treated with various c9, t11-CLA concentrations (25,50,100 and 200 micromol x L(-1)) of c9, t11-CLA for 24 h and 48 h, with a negative control (0.1 % ethanol). RESULTS: The growth of SGC-7901 cells was inhibited by c9,t11-CLA. Eight days after treatment with various concentrations of c9,t11-CLA, as mentioned above, the inhibition rates were 5.9 %, 20.2 %,75.6 % and 82.4 %, respectively. The frequency of apoptosis on SGC-7901 cells induced by different concentrations of c9, t11-CLA (except for 25 micromol.L(-1), 24 h) was significantly greater than that in the negative control (P<0.01). To further investigate the influence of the cell cycle progression, we found that apoptosis induced by c9, t11-CLA may be involved in blocking the cell cycle of SGC-7901 cells. Immunocytochemical staining demonstrated that SGC-7901 cells preincubated in media supplemented with different c9, t11-CLA concentrations for various time periods significantly decreased the expressions of ki67 (the expression rates were 18.70-3.20 %, at 24 h and 8.10-0.20 % at 48 h, respectively), bcl-2 (4.30-0.15 % at 24 h and 8.05 %-0 at 48 h), and c-myc (4.85-2.20 % at 24 h and 4.75-0.30 % at 48 h) as compared with those in the controls (the expressions of ki67, bcl-2, and c-myc were 15.1 % at 24 h and 13.5 % at 48 h, 6.80 % at 24 h and 8.00 % at 48 h, 5.50 % at 24 h and 5.30 % at 48 h, respectively) (P<0.01), whereas the expressions of Fas were increased (0.60-2.75 %, 24 h and 0.45-5.95 %, 48 h). CONCLUSION: The growth and proliferation of SGC-7901 cells are inhibited by c9, t11-CLA via blocking the cell cycle, pathways of bcl-2-associated mitochondria with reduced expression of bcl-2 and Fas-associated death domain protein (FADD) with enhanced expression of Fas. But expression of c-myc on SGC-7901 cells is lower than that in negative control, which needs to be studied further.     

Effect of Nimesulide on proliferation and apoptosis of human hepatoma SMMC—7721 cells

AIM:Cyclooxygenase-2(COX-2)has been suggested to be associated with carcinogenesis.We sought to investigate the effect of the selective COX-2 inhibitor,Nimesulide on proliferation and apoptosis of SMMC-7721 human hepatoma cells.METHODS:This study was carried out on the culture of hepatic carcinoma SMMC-7721 cell line Various concentrations of Nimesulide(0,200μmol/L,300μmol/L,400μmol/L)were added and incubated.Cell proliferation was detected with MTT colorimetric assay,cell proliferation was detected with MTT colorimetric assay,cell apoptosis by electron microscopy,flow cytometry and TUNEL.RESULTS:Nimesulide could significantly inhibit SMMC-7721 cells proliferation dose-dependent and in a dependent manner compared with that of the controla group.The duration lowerst inhibition rate produced by Nimesulide in SMMC-7721 cells was 19.06%,the highest inhibition rate was 58.49%,After incubation with Nimesulide for 72h,the most highest apotosis rate and apoptosis index of SMMC-7721 cells comparing with those of the control were 21.20%&#177;1.62%,vs2.24%&#177;0.26%and 21.23&#177;1.78vs2.01&#177;0.23(P&lt;0.05).CONCLUSION:The selective COX-2 inhibitor,Nimesulide can inhibit the proliferation of SMMC-7721 cells and increase apoptosis rate and apoptosis index of SMMC-7721 cells,The apoptosis rate and the apoptosis index are dose-dependent.Under electron microscope SMMC-7721 cells incubated with 300 μmol and 400μmol Nimesulide show apoptotic characteristics With the clarification of the mechanism of selective COX-2 inhibitors,Thtese COX-2 selective inhibitors can become the choice of prevention and treatment of cancers.     

Effect of cis—9,trans—11—conjugated linoleic acid on cell cycle of gastric adenocarcinoma cell line（SGC—7901）

AIM: To determine the effect of cis -9, trans -11-conjugated linoleic acid (c9, t11-CLA) on the cell cycle of gastric cancer cells (SGC-7901) and its possible mechanism in inhibition cancer growth. METHODS: Using cell culture and immunocytochemical techniques, we examined the cell growth, DNA synthesis, expression of PCNA, cyclin A, B(1), D(1), p16(ink4a) and p21(cip/waf1) of SGC-7901 cells which were treated with various c9, t11-CLA concentrations (25, 50, 100 and 200 micromol.L(-1))of c 9, t 11-CLA for 24 and 48h, with a negative control (0.1% ethane). RESULTS: The cell growth and DNA synthesis of SGC-7901 cells were inhibited by c9, t11-CLA.SGC-7901 cells. Eight day after treatment with various concentrations of c9, t11-CLA mentioned above, the inhibition rates were 5.92%, 20.15%, 75.61% and 82.44%, respectively and inhibitory effect of c9, t11-CLA on DNA synthesis (except for 25 micromol.L, 24h) showed significantly less (3)H-TdR incorporation than that in the negative controls (P<0.05 and P<0.01). Immunocytochemical staining demonstrated that SGC-7901 cells preincubated in media supplemented with different c9, t11-CLA concentrations at various times significantly decreased the expressions of PCNA (the expression rates were 7.2-3.0%, 24h and 9.1-0.9% at 48h, respectively), Cyclin A (11.0-2.3%, 24h and 8.5-0.5%,48h), B(1) (4.8-1.8% at 24h and 5.5-0.6% at 48h)and D(1) (3.6-1.4% at 24h and 3.7%-0 at 48h) as compared with those in the negative controls(the expressions of PCNA, Cyclin A, B(1) and D(1) were 6.5% at 24h and 9.0% at 48h, 4.2% at 24h and 5.1% at 48h, 9.5% at 24h and 6.0% at 48h,respectively)(P<0.01), whereas the expressions of P16(ink4a) and P21(cip/waf1), cyclin-dependent kinases inhibitors(CDKI), were increased. CONCLUSION: The cell growth and proliferation of SGC-7901 cell is inhibited by c9, t11-CLA via blocking the cell cycle, with reduced expressions of cyclin A,B(1) and D(1) and enhanced expressions of CDKI(P16(ink4a) and p21(cip/waf1)).     

HCV—specific cytokine induction in monocytes of patients with different oputcomes of hepatitis C

AIM：Cytokine release by macrophages critically determines the type of immune response to an antigen Therefore.we studied hepatitis C virus (HCV0-Specific induction of interleukins-1β,-10,-12(IL-1β,il-10,IL-12),and tumor necrosis factor-α(TNF-α) in monocytes.METHODS:Intracellular cytokine expression was studied by flow cytometry in 23 patients with chronic hepatitis C,14 anti-HCV seropositives without viremia and 11 controls after stimulation of peripheral blood mononuclear cells with recombinant core,NS3,NS4 NS5a and NS5b proteins .RESULTS:Patients with HCV viremia revealed greater spontaneous exprssion of IL-1β,TNF-α,and IL-10,Furthermore,greater than twofold higher IL-10 epression was induced by the HCV antigens in chronic hepatitis C than in the other two groups (P&lt;0.05) In contrast,neither IL-12 noir TNF-α was induced preferentially.CONCLUSION:In chonic hepatitis C antigen-specific cytokine induction in monocytes is apparently shifted towards predominant IL-10 induction-not counterbalanced by antiviral type 1 cytokines,This may contribute to persistent viral replication.     

Changes of NF—kB,p53,Bcl—2 and caspase in apoptosis induced by JTE—522 in human gastric adenocarcinoma cell line AGS cells：role of reactive oxygen species

AIM:To identify whether JTE-522 can induce apoptosis in AGS cells and ROS also involved in the process,and to investigate the changes in NF-kB,p53,bcl-2 and caspase in the apoptosis process.METHODS:Cell culture,MTT,Electromicroscopy,agarose gel electrophoresis,lucigenin,western blot and electrophoretic mobility shift assay(EMSA)analysis were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanisms.RESULTS:JTE-522 inhitied the growth of AGS cells and induced the apoptosis,Lucigenin assay showed thd generation of ROS in cells under incubation with JTE-522,The increased ROS generation might Contribute to the induction of AGS cells to apoptosis.EMSA and Western blot revealed that NF-kB activity was almost completely inhihbited by preventing the degradation of IKBα,Additionally,by using Western blot we confirmed that the level of bcl-2 was decreased,whtereas p53 showed a great inmcrease following JTE-522 treatment.Their changes were in a dose-dependent manner. CONCLUSION：These findings suggest that reactive oxygen species,NF-kB,p53,bcl-2 and caspase-3 may play an important role in the induction of apoptosis,in ags cells after treatment with JTE-522.     

CCK-8 inhibits expression of TNF-α in the spleen of endotoxic shock rats and sigual transduction mechanism of p38 MAPK

AIM: To study the effect of sulfated cholecystokinin-octapeptide (CCK-8) on systemic hypotension, gene andprotein expression of TNF-α in spleen of lipopolysaccharide(LPS)-induced endotoxic shock (ES) rats, and furtherinvestigate the signal transduction mechanism of p38mitogen-activated protein kinase (MAPK).METHODS: The changes of blood pressure were observedusing physiological record instrument in four groups of rats:LPS(8 mg@kg-1, iv), CCK-8 (40μg@kg-1, iv)pretreatment10min before LPS (8 rmg@kg-1), CCK-8 (40μg@kg-1, iv) ornormal saline (control) group. The content of TNF-αinspleen was assayed 2 h after LPS administration usingELISA kit and the expression of TNF-α mRNA was examined30 min, 2 h and 6 h after LPS administration by reversetranscribed polymerasa chain reaction (RT-PCR). Activationof p38 MAPK was detected with Western blot 30 min afterLPS administration.RESULTS: CCK-8 reversed LPS-induced decrease of meanarterial pressure (MAP) in rets. The content of TNF-α inspleen was (282 ± 30) ng@ L-1 in control group, while itincreased to (941 ± 149) ng@L-1 in LPS group, P＜ 0.01.CCK-8 significantly inhibited the LPS-induced increase ofTNF-α content in spleen. lt decreased to (462 ± 87) ng@ L- 1 inCCK- 8 + LPS group, P ＜ 0.01. The expression of TNF-αmRNA 30 min and 2 h after treatment was stronger in LPSgroup, while it was lowered after CCK-8 pretreatment. Thep38 MAPK expression increased significantly in LPS group(5.84 times of control) and CCK-8 increased the activationof p38 MAPK in ES rats (10.74 times of control).CONCLUSION: CCK-8 reverses the decrease of MAP in ESrats and has inhibitory effect on the gene and proteinexpression of TNF-α in spleen, and p38 MAPK may beinvolved in its signal transduction mechanisms.     

Anti—inflammatory effect of cholecystokinin and its signal transduction mechanism in endotoxic shock rat

AIM：To study the anti-inflammatory effects of choecystokinin-octapeptide(CCK-8)on lipopolysaccharide(LPS)-induced endotoxic shock(ES)and further investigate its signal transduction pathways involving p38mitogen-activated protein kinase(MAPK)andIкα.METHODS:Eighty-four rats were divided randomly into four groups:LPS(8ng&#183;kg^-1,iv)inducedES;CCK-8(40μg&#183;kg^-1,iv)pretreatment10min before LPS(8mg&#183;kg^-1);CCK-8(40μg&#183;kg^-1,iv)ornormal saline (control)groups.The inflammatory changes of lung and spleen,phagocytic function of alveolar macrophage,quantification of inflammatory cells in bronchoalveolar lavage(BAL)were investigated in rats by using hematoxylin and eosin(HE)staining.phagocytosis of Candida albicans and differential cell counting,Nitric oxide(NO)production in serum,lung and spleen was measured with the griess reaction.The mechanism involving p38mapkandIкB-αsignal pathways was investigated by Western blot.RESULTS:Inflammatory changes of lung and spleen induced by LPS were alleviated by CCk-8,the increase of NOinduced by LPSin serum,lung and spleen was significantly inhibited and the neutrophil infiltration in BALwas significantly reduced by CCK-8,The number of neutrophils was(52&#177;10)&#215;10^6cells&#183;L^-1inCCK-8+LPS(P&lt;0.01).The phagocytic rate of CCK-8 group increased to (62.49&#177;9.94)%,compared with control group(48.16&#177;14.20)%,P&lt;0.05,The phagocytosis rate was(85.14&#177;4.64)%in LPSgroup,which reduced to(59.33&#177;3.14)%inCCK-8|LPSgroup(P&lt;0.01).The results of phagocytosis indexes showed similar changes.CCK-8may play an important role in increasing the expression of p38MAPKand decreasing the degradation of IкB-αin lung and spleen of ES rats. effects,which may be related to activation of p38MAPKand inhibition on the degradation of IкB-α.     

Effect of cholecystokinin on cytokines during endotoxic shock in rats

AIM To study the effect of cholecystokinin-octapeptide(CCK-8)on systemic hypotension and cytokine productionin lipopolysaccharide(LPS)-induced endotoxic shock(ES)rats.METHODS The changes of blood pressure were observedusing physiological record instrument in four groups ofrats:LPS(8mg·kg~(-1),iv)induced ES;CCK-8(40μg·kg~1,iv)pretreatment 10min before LPS(8mg.kg~(-1));CCK-8(40μg·kg~(-1),iv)or normal saline(control)groups.Differences in tissue and circulating specificity of theproinflammatory cytokines(TNF-α,IL-1β and IL-6)wereassayed with ELISA kits.RESULTS CCK-8 reversed LPS-induced decrease of meanartery blood pressure(MABP)in rats.Compared withcontrol,LPS elevated the serum level of IL-6 significantly(3567±687ng·L~(-1) vs 128±22ng·L~1,P<0.01),whilecontents of TNF-α and IL-1β elevated significantly(277±86ng.L~1 vs not detectable and 43±9ng·L~1 vs notdetectable,P<0.01)but less extent than IL-6.CCK-8significantly inhibited the LPS-induced increase in serumTNF-α,IL-1β and IL-6.LPS elevated spleen and lungcontent of IL-1β significantly(5184±85ng·L~1 vs 1047±21ng·L~1 and 4050±614ng·L~1 vs not detectable.P<0.01),while levels of TNF-α and IL-6 also rosesignificantly but in less extent than IL-1β.CCK-8 inhibitedthe LPS-induced increase of the cytokines in spleen andlung.In the heart,CCK-8 significantly inhibited LPS-induced increase of TNF-α(864±123ng·L~1 in CCK-8 LPS group vs 1599±227ng·L~1 in LPS group,P<0.01),and IL-1β(282±93ng·L~1 in CCK-8 LPS group vs 621±145ng·L~1 in LPS group,P<0.01).CONCLUSION CCK-8 reverses ES,which may be relatedto its inhibitory effect on the overproduction of cytokines.     

Antioxidative effect of p38 mitogen-activated protein kinase inhibitor in the kidney of hypertensive rat

OBJECTIVE: Nicotinamide adenine dinucleotide phosphate (NAD(P)H) oxidase is regulated by angiotensin II, interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha via p38 mitogen-activated protein kinase (MAPK). We hypothesized that p38 MAPK inhibitor, FR167653, may suppress NAD(P)H oxidase and its oxygen radical production and ameliorate renal damage in Dahl salt-sensitive rats with heart failure (DSHF). METHODS: DSHF rats were fed with 8% NaCl diet from 6 to 18 weeks old. Eleven-week-old DSHF rats received either vehicle or FR167653 (2 mg/kg per day) for 7 weeks and the renal NAD(P)H oxidase p47phox and nitric oxide synthase (NOS), superoxide production and renal damage were evaluated in comparison with the control Dahl salt-resistant rat fed with 8% NaCl diet. RESULTS: In the kidney of DSHF rat, phosphorylated p38 MAPK was enhanced with an increased IL-1beta and TNF-alpha production compared with control rats. Treatment with FR167653 significantly suppressed p38 MAPK, IL-1beta and TNF-alpha. Renal NAD(P)H oxidase p47phox expression and superoxide production were significantly increased in the DSHF rats and treatment with FR167653 suppressed NAD(P)H oxidase expression and reduced superoxide formation. Renal endothelial and inducible NOS were reduced in DSHF rats compared with control rats, but FR167653 increased NOS and NO production in the kidney. Proteinuria, glomerulosclerosis and interstitial macrophage migration via intercellular adhesion molecule-1 (ICAM-1) were enhanced in DSHF rat and they were ameliorated by FR167653. CONCLUSION: The inhibition of p38 MAPK by FR167653 reduced renal IL-1beta and TNF-alpha production and ameliorated renal damage in hypertensive rat via suppression of NAD(P)H oxidase and enhanced NO bioavailability.     

p38丝裂原活化蛋白激酶对急性坏死性胰腺炎大鼠低钙血症的影响

目的 探讨p38丝裂原活化蛋白激酶(p38MAPK)信号转导通路对急性坏死性胰腺炎(ANP)大鼠低钙血症和甲状旁腺激素受体1(PTHR1)表达的影响.方法 将雄性SD大鼠72只按完全随机法分为ANP组、SB203580干预(SB)组和假手术(SO)组,每组分3、6、12 h 3个时间点,每个时间点8只.以5%牛磺脱氧胆酸钠逆行胰胆管注射建立ANP模型,SB组在造模前30 min腹腔注射p38MAPK特异抑制剂SB203580 10 mg/kg体重.观察各组血清钙浓度,蛋白质印迹法(Western blotting)分析骨组织磷酸化p38MAPK(P-p38 MAPK)和TNF-α变化,实时RT-PCR检测骨组织PTHR1 mRNA表达.结果 制模后6 h,SO组、ANP组和SB组血清钙浓度分别为(2.50±0.08)mmoL/L、(2.11±0.06)mmol/L和(2.35±0.10)mmol/L;骨组织P-p38 MAPK表达量分别为0.14±0.04、0.80±0.06和0.33±0.05;骨组织TNF-α表达量分别为0、0.91±0.04和0.44±0.03;骨组织PTHR1 mRNA表达量分别为1.00±0.12、0.23±0.04和0.44±0.06.SB组骨组织P-p38 MAPK及TNF-α表达较ANP组显著降低(P＜0.01);骨组织PTHR1 mRNA表达量及血清钙浓度较ANP组显著增加(P＜0.01).结论 p38MAPK信号转导通路可介导ANP低钙血症的发生,抑制该通路可改善ANP低钙血症.     

Activated protein C,an anticoagulant polypeptide,ameliorates severe acute pancreatitis via regulation of mitogen-activated protein kinases

Background Our aim was to investigate the changes of mitogen-activated protein kinases (MAPKs) by activated protein C (APC) treatment in rats with severe acute pancreatitis (SAP), and relate them to changes in SAP severity, thus providing evidence for developing clinical therapies. Methods Sprague-Dawley rats were given an intravenous injection of saline (SAP group), APC (50 μg/kg or 10 μg/kg), or CNI1493 just before SAP induction. One group of rats underwent a sham operation (control group). Experimental samples were harvested 16 h after SAP induction. The gene expression of pancreatic MAPKs was evaluated by cDNA microarrays. The mRNA and protein/phosphorylated protein levels of p38 MAPK, extracellular signal-regulated protein kinase (ERK) 1/2, and c-Jun N-terminal kinase (JNK) and the protein levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β were determined in pancreatic tissue. The severity of disease was evaluated by pancreatic histology, the pancreatic wet/dry weight ratio, and the serum amylase level. Results In rats treated with APC (50 μg/kg) or CNI1493, the severity of pancreatitis and expression of pancreatic TNF-α and IL-1β proteins were attenuated by the decreased expression and activity of p38 MAPK and JNK (vs. the SAP group, P < 0.01). The expression and activity of ERK1/2 were increased in APC-treated rats, especially in the group treated with APC 50 μg/kg (vs. the SAP or CNI1493-treated group, P < 0.01, respectively). Conclusions Inhibition of expression of pancreatic p38 MAPK and JNK and upregulation of ERK1/2 expression by APC treatment may protect against pancreatic injury, thus ameliorating severity of the disease.     

Effect of atrial natriuretic peptide on ischemia-reperfusion injury in a porcine total hepatic vascular exclusion model

AIM： To evaluate the effect of ANP on warm I/R injury in a porcine THVE model.
METHODS： Miniature pigs （mini-pigs） weighing 16-24 kg were observed for 120 min after reperfusion following 120 min of THVE. The animals were divided into two groups. ANP （0.1 μg/kg per min） was administered to the ANP group （n = 7）, and vehicle was administered to the control group （n = 7）. Either vehicle or ANP was intravenously administered from 30 min before the THVE to the end of the experiment. Arterial blood was collected to measure AST, LDH, and TNF-α. Hepatic tissue blood flow （HTBF） was also measured. Liver specimens were harvested for p38 MAPK analysis and histological study. Those results were compared between the two groups.
RESULTS： The AST and LDH levels were lower in the ANP group than in the control group; the AST levels were significantly different between the two groups （60 min： 568.7 ± 113.3 vs 321.6 ± 60.1, P = 0.038 〈 0.05, 120 rain： 673.6± 148.2 vs 281.1±44.8, P = 0.004 〈 0.01）. No significant difference was observed in the TNF-α levels between the two groups. HTBF was higher in the ANP group, but the difference was not significant. A significantly higher level of phosphorylated p38 MAPK was observed in the ANP group compared to the control group （0min： 2.92± 1.1 vs 6.38 ±1.1,,P= 0.011 〈 0.05）.
Histological tissue damage was milder in the ANP group than in the control group.
CONCLUSION： Our results show that ANP has a protective role in I/R injury with p38 MAPK activation in a porcine THVE model.