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Objective To investigate the effect of cytokines on pancreatic function in patients after acute pancreatitis(AP) and its mechanisms. Methods Fifty-nine patients (mild in 25 and severe in 34) after AP and 20 healthy controls were enrolled in the study. Serum levels of cytokines including hepatocyte growth factor (HGF), epidermal growth factor(EGF), basic fibroblast growth factor (bFGF), regeneration protein(Reg)-1 and Reg-4 were determined using enzyme-linked immunosorbent assay (ELISA). Fasting blood-glucose, insulin, C-peptide and fecal elastase 1 (FE1) were detected for evluation of endocrine and exocrine pancreatic function. The association of pancreatic function with clinical parameters and serum cytokines was analyzed. Results The expression of FE1 was lower in patients [(205.9±18.3) μg/g] after AP in comparison with the controls [(333.9±19.7) μg/g, P<0. 01], but levels of fasting blood-glucose, C-peptide and insulin were higher in patients group (P<0.01). Serum level of HGF was higher in patients with insufficient pancreatic exoerine [(983.76±372.65) pg/ml] than those with normal exocrine function [(263.44±110. 35) pg/ml]. Meanwhile,EGF level was higher in patients with DM after AP [(704.41±190. 37) pg/ml] than those without DM [(360. 03±48.39) pg/mh P<0.05]. There was a negatively correlation between FE1 and HGF (P ＜0. 01). The abnormal fasting blood glucose was correlated with CT grading (P<0. 05).Conclusions The patients after AP develope insufficient exocrine and endocrine function. Serum EGF and HGF may be associated with restoration of pancreatic endocrine and exocrine function.
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Objective To investigate the role of Hedgehog pathway in hepatic fibrosis and its association with activation of hepatic stellate cells. Methods Twenty male Spragur-Dawley rats were divided into control and model groups with 10 each. The animal models were induced by injection with CCl4 and fed with fat-rich diet. The rats in both groups were sacrified at the 8 week with 5 each and the liver tissues were removed for HSC-T6 culture. The deposition of collagen fiber in liver was detected with HE and Masson staining. RT-PCR was used to detect the expressions of Sonic hedgehog (Shh), smoothened (Smo), patched (Ptc), Gli-1 and α-smooth muscle action (α-SMA) mRNA in HSC-T6 and liver tissues. The influence of cyclopamine (Cyc) and lipopolysaccharide (LPS) on HSC-T6 proliferation were assayed by MTT. The expressions of Shh, Smo, Ptc, Gli-1 and α-SMA mRNA after intervention with Cyc (100μmol/L) and LPS were measured by real-time PCR. Results A lot of lipo and collagen deposited in liver of model rats. The Shh,Smo,Gli-1 and α-SMA mRNA were highly expressed in model rats than those in control group (2-△△Ct were 20.45±3.31 vs. 1, 12.78 ± 0. 53 vs. 1, 10.88 ± 2.41 vs. 1, 4.91 ± 2. 59 vs. 1, respectively, all P value <0. 05). In vitro Cyc inhibited HSC-T6 proliferation in dose dependant manner (F=636.81, P<0.01). Compared to the control group, the mRNA expressions of Smo, Ptc, Gli-1,α-SMA in HSC-T6 were significantly reduced after Cyc intervention (2△△Ct, were 0. 20±0. 11, 0. 21 ± 0. 08, 0. 28 ± 0. 05,0. 27±0.10,respectively, all P values<0.01). Conclusion The expression of members of Hedgehog pathway are increased in the progress of hepatic fibrosis, which may accelerate the hepatiee fibrosis by activating HSC.
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Objective Some experimental results indicated that cyclooxygenase 2 (COX 2) was involved in the regulatory process of balance between cell apoptosis and multiplication. The present study was designed to investigate the effect of COX 2 on the course of pancreatic adenocarcinoma cell apoptosis and its possible signal pathway. Methods Apoptosis of pancreatic cancer cells (PC 3, highly expressed COX 2) induced by selective COX 2 inhibitor, Celebrex (IC 50 , 100 ?mol), was detected by using DNA gel electrophoresis, flow cytometry and electron microscopy. Expressions of apoptotic related genes mRNA, including bcl xl, bax, Survivin, were analyzed by reverse transcription polymerase chain reaction (RT PCR). Results Substantial apoptosis was induced by the treating PC 3 cells with Celebrex, as revealed by typical ladder pattern of DNA fragments under DNA electrophoresis, increment of apoptotic apportion, and apoptotic body under electron microscopy. Apoptotic inhibitory genes, bcl xl, bax, Survivin, were expressed in PC 3 cells, and were down regulated significantly by Celebrex in bcl xl and Survivin but not for bax. Conclusion Above findings suggest that the COX 2 pathway contributes to the apoptosis of pancreatic cancer cell, which may be via signal transduction of bcl xl and Survivin genes.
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Objective To investigate the deleterious effects of enterogenous endotoxemia on the course of acute pancreatitis in mice, and its possible mechanisms. Methods Sixty five C57BL/6 mice were assigned to 4 groups randomly, including acute edematous pancreatitis (AEP), lipopolysacchride (LPS), AEP plus LPS and normal control. AEP was induced by intraperitoneal injection of cerulein with a dosage of 50 ?g/kg at hourly interval for seven times under ether anesthesia. LPS was administrated via a gastric tube with a dosage of 5 mg/kg at 6 h after the first cerulein or saline injection. Serum amylase and LDH activities were measured at 12 h, 24 h, 48 h and 5 d. Pathological alteration in pancreas was studied. Expressions of Mac 1 (CD11b/CD18), P selectin, E selectin and ICAM 1 were evaluated by using inmmunohistochemical procedures. Expressions of cytokine genes were determined by means of RT PCR and Southern blot. Myeloperoxidase (MPO) activity in pancreas was also analyzed. Results Cerulein induced a typical changes of AEP in mice, which was confirmed by pathological changes and hyperamylasemia. LPS alone didn't develop either morphological changes or biochemical alterations. However, cerulein induced AEP challenged by LPS could cause marked parenchymal necrosis and hemorrhage with significant increment of serum amylase and LDH activities. Expressions of Mac 1, P selectin, E selectin and ICAM 1 in pancreas were enhanced. Cytokine genes including TNF?,IL 1?,IL 6 and IFN? mRNA were also upregulated. MPO activity was increased. Conclusion This study suggested that enterogenous endotoxemia, which could not induce pancreatic injury per se , could induce AEP into ANP in mice. Over stimulation of neutrophils and releasing of pro inflammatory mediators might be the contributing factors.
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AIM: To investigate the effects of the dominant-negative I?B? plasmid on the expression of NF-?B and cyclooxygenase-2 (COX-2) after its being transfected into pancreatic carcinoma PC-3 cell line. METHODS: The expression of NF-?B and COX-2 in PC-3 cell line was confirmed by immunohistochemistry. The effects of dominant-negative I?B? plasmid transfection on the expression of NF-?B and COX-2 were studied by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. RESULTS: Both NF-?B and COX-2 were expressed in pancreatic carcinoma PC-3 cell line, and the expression of NF-?B and COX-2 were down-regulated in a certain time-independent manner after dominant-negative I?B? plasmid transfection. CONCLUSIONS: The NF-?B and COX-2 are expressed in pancreatic carcinoma PC-3 cell lines. The expression of NF-?B and COX-2 are inhibited by dominant-negative I?B? plasmid, while NF-?B is likely to play an important role in regulating the expression of COX-2.
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Objective To investigate the effects and mechanism of Safflower Injection on the proliferation,apoptosis,and expression of bcl-2/bax gene in hepatic stellate cell(HSC) in vitro.Methods HSC Line HSC-T6 was incubated with Safflower Injection at different concertration,cell proliferation was assessed by MTT colorimetric assay.After incubated with Safflower Injection 5,10,and 20 mg/mL,flow cytometry(FCM) was used to detect the content of DNA in HSC-T6.Morphological change of HSC-T6 was observed under microscope and agarose gel electrophoresis for DNA Ladder was used to detect apoptosis.Besides,the early stage of apoptosis was detected with Annexin V-FITC/PI double labbled assay.And real time RT-PCR was used to detect the expression of bcl-2/bax gene.Results The significant inhibition of Safflower Injection on HSC proliferation was observed in a dose-and time-dependent manner.Observed by FCM,the cell ratio in G0/G1 phase with Safflower Injection treatment(10 and 20 mg/mL) for 24 h was increased,which showed the significant difference compared with the control group(P