【 Abstract 】 Objective Overexpressions of epidermal growth factor (EGF) and EGF receptor have been associated with progression and invasive phenotype of pancreatic cancer. However, the underlying molecular mechanism by which EGF worked in pancreatic cancer cells has not been completely understood. In this study, effect of EGF on the invasion and metastasis of pancreatic cancer cells and its regulatory mechanism were investigated. Methods The effects of EGF on the proliferation, adhesion and invasion of pancreatic cancer cells were detected by WST-1 proliferation assay, adhesion assay and invasive assay, respectively. The activity and expression of MMP-2 and MMP-9 were examined by zymography, Western blot and RT-PCR, respectively. The activity of NF- κ B was examined by EMSA. Results EGF could significantly promote the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion. The expressions of NF- κ B and MMP-9 were significantly increased by EGF, but EGF did not affect the activity and expression of MMP-2. Furthermore, EGF stimulated the NF- κ B binding activity. Pretreatment with NF- κ B inhibitors, pyrrolidine dithiocarbamate (PDTC), could significantly inhibit the activity of NF- κ B induced by EGF. Meanwhile, the EGF-induced expression and activity of MMP-9, as well as cell invasiveness were also inhibited by NF- κ B inhibitor. Conclusion EGF could increase the expression and promote the invasiveness of MMP-9 via the activation of NF- κ B in pancreatic cancer cells, which implies that NF- κ B inhibitant, such as PDTC, may diminish the invasiveness of pancreatic cancer cells.
ObjectiveTo investigate the possible protective effect of zerumbone on pancreatic injury in severe acute pancreatitis (SAP) rats, and to provide a theoretical basis for prevention and treatmen of severe acute pancreatitis. MethodsSeventy male Wistar rats were randomly divided into four groups:normal control group (NC group, n=10), SAP group (n=40), Zerumbone pretreatment group (ZER group, n=10), and Zerumbone drug control group (ZER-CON group, n=10). Rats of SAP group were divided into four time points of 1 h, 3 h, 6 h, and 12 h (n=10 each time point). SAP models were induced by retrograde injection of 5% sodium taurocholate (0.1 mL/100 g) in biliopancreatic duct in SAP group and ZER group. Rats were injected isotonic saline solution instead of taurocholate as a control in NC group and ZER-CON group. Zerumbone solution (10 mg/kg) was administered via femoral vein half an hour prior to establishing models in ZER group and ZER-CON group. All rats except SAP group were sacrificed at 12 h time point after the induction of SAP. The rats in SAP group were sacrificed at 1 h, 1 h, 3 h, 6 h and 12 h time point after induction of SAP. The mortality, ascites, serum amylase (AMY), phospholipase, and pathological examination of pancreas were observated or detected. The expression of nuclear factor-kappa B (NF-κB) p65 in pancreatic tissues was evaluated by immunohistochemistry assay. ResultsThere were no difference of the levels of mortality, ascites, serum AMY, phospholipase, pathological examination, and NF-κB p65 location expression of pancreas between the ZER-CON group and NC group (P>0.05). The above indexes in SAP group were significantly higher than those in NC group (P<0.05). However, those in ZER group were significantly lower than in SAP group (P<0.05), and higher than in NC group (P<0.05). ConclusionsZerumbone can reduce the mortality and ascites, effectively alleviate the enzyme, pathological injury, and NF-κB p65 location expression in pancreatic tissue following SAP. It may indicate that zerumbone can protect pancreatic injury in SAP via NF-κB pathway.
ObjectiveTo discuss the role of nuclear factor-kappa B in restenosis after angioplasty.MethodsRelated literatures of recent 5 years were reviewed.ResultsNuclear factor-kappa B could lead to hyperplasia of vascular intima which resulted from proliferation and decrease of apoptosis of vascular smooth muscle cells.ConclusionNuclear factor-kappa B plays an important role in restenosis after angioplasty.
ObjectiveTo investigate the role and mechanism of P-selectin glycoprotein ligand-1 (PSGL-1) in hydrochloric acid-induced acute lung injury (ALI) in mice.MethodsWild-type mice (WT) and PSGL-1 knockout mice (PSGL-1 -/-) were randomly subjected to normal saline (NS) or hydrochloric acid (HCl) challenged group. The mice were intratracheally instilled with NS or HCl (1 μl/g weight) into the left lung with a catheter. After 2 hours, respiratory function index enhanced pause (Penh), PaO2 and PaO2 were analyzed. The wet to dry weight ratio (W/D) of the left lung and total protein concentration in bronchoalveolar lavage fluid (BALF) were measured. The number of leukocytes in BALF was counted too. Targeted lung tissue was processed for further HE or immunohistochemistry staining. Meanwhile, the expressions of interleukin-6 (IL-6), IL-1β, nuclear factor-κB (NF-κB), IκBa and p-IκBa in lung tissue were measured.ResultsThe Penh (4.77±1.22 vs. 5.80±0.84) and PaCO2 [(63.7±3.9) mm Hg vs. (74.4±7.4) mm Hg] in the PSGL-1 knockout mice were significantly lower than those in the WT mice after HCl stimulation (P<0.05), while the PaO2 was higher than that in the WT mice [(81.0±7.1) mm Hg vs. (62.0±8.9) mm Hg, P<0.05)]. The lung W/D ratio (4.86±0.15 vs. 5.22±0.20), protein concentration [(3.71±0.64) μg/μl vs. (4.74±0.98) μg/μl] and total leukocyte count [(13.00±2.18) ×107/L vs. (49.42±3.35) ×107/L] in BALF were significantly lower in the PSGL-1 knockout mice challenged with HCl than those in the WT mice (P<0.05). Besides, the protein expressions of IL-6, IL-1β, p65 and p-IκBa in the PSGL-1 knockout mice were lower than those in the WT mice after HCl instillation, while the IκBa expression was higher than that in the WT mice (P<0.05). More numbers of neutrophils and macrophages were found in the lung of the WT mice than the PSGL-1 knockout mice challenged with HCl. However, the differences of above values between the WT mice and the PSGL-1 knockout mice instilled with NS were not found, all of which were significantly lower than the correspongding HCl group except for IκBa (P<0.05).ConclusionPSGL-1 may play important roles in the development of HCl-induced ALI via the NF-κB signaling pathway and inflammation.
Objective To observe the effects of exogenous pulmonary surfactant (PS) on ventilation-induced lung injury (VILI) in rats, and to investigate its possible mechanisms. Methods A total of 40 Wistar rats were divided into 4 groups with randomized blocks method: control group, high tidal volume (HV) group, VILI group, and PS group, with 10 rats in each group. The control group was subjected to identical surgical procedure but was never ventilated. After 30 min of mechanical ventilation (MV) with Vt 45 ml/kg, the rats in HV group were killed immediately; rats in the VILI group were continually ventilated for up to 150 min with Vt 16 ml/kg; in the PS group, 100 mg/kg of PS administered intratracheally and with the same settings as VILI group. Mean artery pressure (MAP), blood gas analysis, lung wet to dry weight ratios (W/D), thorax-lung compliance, and cell counts in bronchoalveolar lavage fluid (BALF) were determined. Nuclear factor-κB(NF-κB) activity in lungs was measured by enzyme-linked immunosorbent assay (ELISA), interleukin-8(IL-8) in serum and BALF was determined by radioimmunoassay (RIA). Pathological examination of the lung was performed. Results Injurious ventilation significantly decreased MAP and PaO2/FiO2, but increased NF-κB activity and W/D. MAP and PaO2/FiO2 improved, but NF-κB activity, IL-8 in serum and BALF, and cell counts in BALF reduced significantly in PS group compared with those in VILI group. Histological studies showed reduced pulmonary edema and atelectasis in the PS group. Conclusion PS administered intratracheally can suppress the increased activity of NF-κB induced by VILI, exogenous PS can be used to treat VILI.
Abstract: Objective To study the preventive effect of n-3 polyunsaturated fatty acids on allograft arteriosclerosis. Methods Arterial homeotransplant model were created with 480 rats which were divided into four groups. Control group, no n-3 lyunsaturated fatty acids were taken. Group A, n-3 polyunsaturated fatty acids were taken for two weeks before operation with the dose of EPA 600mg/kg. Group B, 300 mg/kg and group C 150 mg/kg were taken respectively. The recipient’s transplanted vessel was excised after 1,7,14,21and 28 days respectively. The tissue pathological variations, ultrastructure variations and expression variations of intercellular adhesion molecule-1(ICAM-1), vascular cell adhesion molecule-1（VCAM-1）, nuclear factorkappa B（NF-κB） had been observed. Results The pathological changes occurred 7 days after operation in control group and were most prominent on the 28th day, blood vessels were obstructed and the expressions of ICAM-1, VCAM1,NF-κB were markedly intensified than those of group A, B, C (Plt;0.05). The pathological variations of transplanted vessel in group A, B, C occurred later than those in control group. The nonobstruction rates in group A, B, C were better than that in control group. The expressions of ICAM-1， VCAM-1， NF-κB in control group were ber than those in group A， B， C (Plt;0.05). The expressions of ICAM-1， VCAM-1， NF-κB after 1 day or 7 days demonstrated no statistically significant change in group A， B， C （Pgt;0.05）. The preventive effect for allograft vessel atheromatosis in group A and group B was ber than that in group C after 14, 21 and 28d (Plt;0.05). There were no significant difference between group A and group B (Pgt; 0.05). Conclusion The n-3 polyunsaturated fatty acids can prevent the allograft vessel atheromatosis, the most effective dose of n-3 polyunsaturated fatty acids is 300 mg/kg.
Objective To observed the effect of IL-1β on expression of caudal-related homeobox gene 1 (CDX1) mRNA and mucoprotein 2 (MUC2) mRNA in cultured human gastric epithelial cells GES-1, and to investigate the underlying signal transduction pathways. Methods ①GES-1 cell was activated with IL-1β of different concentrations and time, the expression levels of CDX1 mRNA and MUC2 mRNA were detected by using real-time PCR. ②GES-1 cell was pretreated with PDTC, a NF-κB inhibitor, for 1 h prior to the addition of IL-1β, then the expressions of CDX1 mRNA and MUC2 mRNA were measured. Results Both CDX1 mRNA and MUC2 mRNA were not examined in GES-1 cell under normal culture conditions. But they could be induced by IL-1β with a dose-dependent manner in a concentration range (P＜0.05); 8 h after treatment with IL-1β, the peak values of the expression levels of CDX1 mRNA and MUC2 mRNA were reached (P＜0.05), then declined gradually. When pre-incubated with NF-κB inhibitor PDTC, the expression levels of CDX1 mRNA and MUC2 mRNA were significantly decreased (P＜0.05). Conclusion IL-1β significantly induces the expressions of CDX1 mRNA and MUC2 mRNA in cultured human gastric epithelial cell GES-1 through the NF-κB signal pathway, which indicates that IL-1β plays a role in the process of intestinal metaplasia.
Objective To explore effects of macrophage migration inhibitory factor (MIF) inhibitor ISO-1 on intestinal injury in acute necrotic pancreatitis in pregnancy (ANPIP) rat. Methods Twenty-four pregnant SD rats were randomly averagely divided into three groups: a sham operation (SO) group, an ANP group, and an ANP model plus ISO-1 treatment group (ISO-1 group). A rat model of ANP was induced by the retrograde injection of 5% sodium taurocholate into the biliopancreatic duct. The rats were killed and the inferior vena cava blood and the tissues of pancreas and jejunum were harvested at 12 h after the operation. The serum amylase (AMY), lipase (LIP), diamine oxidase (DAO), interleukin (IL)-1β, and IL-6 levels were measured. The pancreatic and jejunal tissues were taken for the pathological examination scoring. The immunohistochemical method was used to detect the expression of the MIF, nuclear factor (NK)-κB, or tumor necrosis factor (TNF)-α protein. Results ① Compared with the SO group, the serum AMY, LIP, DAO, IL-1β, and IL-6 levels were increased in the ANP group (P<0.050), which in the ISO-1 group were decreased as compared with the ANP group, the DAO, IL-1β, and IL-6 levels had significant differences (P<0.050), but the AMY and LIP levels had no significant differences (P>0.050). ② The pathological points of the pancreas and jejunum tissues were increased in the ANP group as compared with the SO group, which were significantly decreased in the ISO-1 group as compared with the ANP group (P<0.050). ③ The average integrated optical density divide by area of the NF-κB,TNF-α, and MIF were significantly increased in the ANP group as compared with the SO group, which were significantly decreased in the ISO-1 group as compared with the ANPgroup (P<0.050). Conclusions MIF inhibitor ISO-1 could protect intestinal injury in ANPIP rat. It is suggested that MIF is one of mechanisms in ANPIP with intestinal injury and might be correlated with activities of TNF-α and NF-κB.
ObjectiveTo explore the protective mechanism and effect of the resveratrol for kidney injury of obstructive jaundice. MethodsThe rats were randomly divided into three groups: sham operation group receiving laparotomy without bile duct ligation (BDL), the obstructive jaundice group with BDL, and the obstructive jaundice + resveratrol group given resveratrol following BDL. The levels of total bilirubin (TBIL), direct bilirubin (DBIL), blood urea nitrogen (BUN), and creatinine (Cr) in the serum were tested. The superoxide dismutase (SOD) activity, malondialdehyde (MDA) content, glutathione (GSH) level in the renal tissues were detected. The expressions of the silent information regulator 1 (SIRT1) and nuclear factor-κB (NF-κB) proteins were tested by Western blot. The expression of SIRT1 mRNA was detected by RT-PCR and the renal cell apoptosis was examined by TUNEL staining. Results①Compared with the sham operation group, the levels of serum TBIL, DBIL, BUN and Cr were significantly higher (P < 0.05); the activity of SOD and level of GSH, and the expressions of SIRT1 mRNA and SIRT1 protein in the renal tissues were signi-ficantly lower (P < 0.05); the content of MDA, the expression of NF-κB protein, and the rate of cell apoptosis in the renal tissues were significantly higher (P < 0.05) in the obstructive jaundice group.②Compared with the obstructive jaundice group, the levels of serum TBIL, DBIL, BUN and Cr were significantly lower (P < 0.05); the activity of SOD and level of GSH, and the expressions of SIRT1 mRNA and SIRT1 protein in the renal tissues were significantly higher (P < 0.05); the content of MDA, the expression of NF-κB protein, and the rate of cell apoptosis in the renal tissues were significantly lower (P < 0.05) in the obstructive jaundice+resveratrol group. ConclusionThe resveratrol could alleviate renal damage and play a beneficial role to resist inflammation, oxidation, and apoptosis by activating the SIRT1 which probably inhibits the expression of NF-κB protein and promotes the activity of SOD in cholestatic kidney injury.
Objective To study the expression of nuclear factor-κBp65 (NF-κBp65) in gastric carcinoma and its relationship with vascular endothelial growth factor (VEGF). Methods The expression of NF-κBp65 and VEGF in 56 gastric carcinomas was detected with immunohistochemistry and compared with benign tissues. Results The positive rates of NF-κBp65 and VEGF in 56 gastric carcinomas were 62.5% and 76.8% respectively,and which were higher than those of gastric mucosal atypical hyperplasia (33.3% and 44.4%) and the normal gastric mucosa(0 and 8.3%) （P＜0.05,P＜0.01）.It was found that there was relationship between the expression of NF-κBp65 and the clinical stage, invasion depth of tumor and lymph node metastasis (P＜0.05),but there was no relation to the historica type (Pgt;0.05). There was positive correlation between NF-κBp65 and VEGF expression （r=0.36,P＜0.01）. Conclusion NF-κBp65 may play an important role in the development of gastric carcinoma by up-regulate the expression of VEGF.