The effect of smoking on endothelin-1 in patients with chronic pancreatitis.

The aim of this study is to prove the influence of tobacco smoking on the endothelin-1 (ET-1) level in the plasma and on the immunohistochemical localization in the pancreatic tissues. The blood was collected from 50 healthy individuals and 63 patients with chronic pancreatitis (CP). The ET-1 and cotinine concentrations in the plasma were estimated by ELISA. Samples of tissues of the normal pancreas and CP were verified histopathologically, and then ET-1 was localized by immunohistochemical staining using the monoclonal anti-human ET-1 antibody. The intensity of immunohistochemical reaction was calculated with the semiquantitative Digital Imaging Methodology. The study demonstrated a significant concentration of ET-1 in smoking healthy individuals and in patients with CP when compared with the nonsmoking population (P=0.003 and 0.0005, respectively). A significantly stronger immunohistochemical ET-1 reaction was observed in the tissue of smoking patients with CP than in the normal pancreatic tissue and of nonsmoking CP patients (P=0.001, 0.008, and 0.03, respectively). The presented data evidence that tobacco smoking has a direct effect on the endothelium, leading to an increased level of ET-1.

Plasma endothelin-1 levels in pancreatic inflammations.

BACKGROUND: In inflammatory processes endothelin exerts an important effect on the pancreatic blood vessels and disturbances in pancreatic microcirculation. OBJECTIVES: The aim of the study was to evaluate the usefulness of plasma endothelin-1 (ET-1) levels in determining the severity of pancreatitis, its prognosis and monitoring the course of therapy. MATERIAL AND METHODS: The investigations were carried out on 115 subjects: 85 patients with pancreatitis and 30 persons serving as a control group. All the participants' ET-1 levels were determined by means of an enzymoimmunological test (Endothelin ETA Kit, Catalog No:583151, Cayman Chemical Company, Michigan, USA) on the 1st, 3rd, 5th and 7th days of treatment. In the control group, the mean ET-1 level was 1.37 pg/ml. RESULTS: Acute pancreatitis (AP) was diagnosed in 53 patients, including 36 patients (67.9%) with severe AP. Their Ranson's scores were over 3 points. The highest mean plasma ET-1 levels were observed in 10 patients with severe AP with necrosis (6.37 +/- 1.9pg/ml). (5 to 9 points on Ranson's scale). In 26 patients with severe AP without necrosis, the mean plasma ET-1 level was 3.49 +/- 1.24 pg/ml. In 17 patients with mild AP the mean plasma ET-1 level was 3.48 +/-1.16 pg/ml (3 to 4 points on Ranson's scale). ET-1 levels normalized gradually with successful treatment. An increase in ET-1 levels between the 5th and 7th days was an unfavorable prognostic factor that was observed in all the patients who died. CONCLUSIONS: Measuring plasma ET-1 levels permits early determination of cases with severe AP. ET-1 can be used as a marker for both the progress of the disease and the efficacy of the treatment. An increase in plasma ET-1 level between the 5th and 7th days of treatment may indicate irreversible ischemic lesions in the pancreas and the development of necrotic lesions.

The effect of smoking on expression of IL-6 and antioxidants in pancreatic fluids and tissues in patients with chronic pancreatitis.

OBJECTIVES: The present study was conducted to ascertain whether cigarette smoke induces oxidative stress in the human pancreas concurrently with inflammation. MATERIALS AND METHODS: The influence of tobacco smoking on the serum level of interleukin-6 (IL-6), on the activities of glutathione peroxidase (GPx) and copper-zinc superoxide dismutase (Cu/Zn SOD) as well as on the metallothionein (MT) level in the pancreatic pseudocyst fluid and its immunohistochemical localization in tissues of non-smoking (n = 9) and smoking (n = 12) patients with diagnosed chronic pancreatitis (CP) was measured. The concentration of interleukin-6 and metallothionein was determined by means of ELISA and the radioisotopic method, respectively. The enzyme activities in the fluid were assayed by the colorimetric method. Samples of tissues of normal pancreas (n = 4) and CP (non-smoking n = 7; smoking n = 12) were verified histopathologically and then IL-6, MT and enzymes were localized by immunohistochemical staining using the monoclonal anti-human antibody. RESULTS: The concentrations of metallothionein and interleukin-6 were significantly higher in smoking patients with CP (as compared with the non-smoking population (p < 0.01; p < 0.001). Interestingly, the ratio of MT/IL-6 in smoking patients with CP was reduced in comparison to non-smoking patients (respectively: 1.1; 5.6). In smoking patients, a significant elevation of the Cu/Zn SOD and GPx activities was revealed as compared with the non-smokers (p < 0.04; p < 0.0007). These studies clearly demonstrate a moderate and strong expression of IL-6 and enzymes in acinar, islet and duct cells of smoking patients. CONCLUSIONS: These observations favor the role of the oxidative stress in the induction of pancreatitis associated with chronic cigarette smoke inhalation.

Immunohistochemical localization of interleukin-6 in human pancreatitis.

The aim of the study was to identify immunohistochemically the localization of interleukin (IL)-6 in normal pancreas and in chronic pancreatitis (CP). Samples of tissues of normal pancreas (n=5) and CP (n=16), were verified histopathologically and then IL-6 was localized by immunohistochemical staining using the monoclonal antihuman IL-6 antibody and test LSAB2-HRP to visualize IL-6/Ab complexes. In slices of the pancreas, derived from patients with CP, a much stronger immunohistochemical reaction was noticed as compared with controls specimens. IL-6 was localized in exocrine, islet cells and ducts cells of the pancreas. Interestingly, this cytokine was detected in cytoplasm and very close to nucleus. Moreover, in cases of CP with inflammatory infiltration, there were a markedly stronger IL-6 expression, than that observed in specimens without infiltrate. In conclusion, the results presented herein clearly demonstrated a moderate and strong expression of IL-6 in exocrine and endocrine cells of patients with CP. These observations provide further support for the existence of local immune-pancreatic interactions.

Blood plasma antioxidant defense in patients with pancreatitis.

OBJECTIVES: The aim of this study was to establish the main determinants of plasma antioxidant potential in different types of human pancreatitis: chronic pancreatitis (CP, n = 19), chronic pancreatitis exacerbated (CPE, n = 16), and acute pancreatitis (AP, n = 8). METHODS: We measured the plasma thiobarbituric acid reactive substances (TBARS), total peroxyl radical trapping potential (TRAP), and the levels of antioxidants such as uric acid (UA), SH groups (SH), ascorbic acid (AA), and bilirubin (BIL). RESULTS: In comparison to healthy volunteers (HV, n = 22) the average levels of TBARS in all types of pancreatitis were significantly higher (by 50%-90%), and plasma SH groups were lower (approximately 30%). In the multivariate regression models, the variability of plasma TRAP was explained in the HV group through UA only (36%), in the CP group through UA, AA, and BIL together (75%), and in CPE group through UA and AA together (81%). Plasma SH concentration did not enter the regression models in all groups. The AP group revealed "paradoxically" high concentrations of AA and TRAP and a negative, linear correlation between the level of TRAP versus UA concentration (R = -0.87). CONCLUSIONS: The multivariate regression analysis points to different structures of antioxidant defense in healthy people and those with pancreatitis: they were much the same in the CP and CPE groups but fundamentally different in the AP group. AP may activate unidentified antioxidant defenses as an adaptive response against oxidative stress.

Immunohistochemical localization of metallothionein in chronic pancreatitis.

Metallothionein (MT) is a low-molecular weight intracellular protein, rich in sulfhydryl residues, and able to bind bivalent metals. MT, like Zn, is a component of the diversified elements of antioxidant system. Recent studies have shown that reactive oxygen species play a role in the pathogenesis and development of chronic pancreatitis. The aim of the study was to identify immunohistochemically (LSAB2-HRP; DAKOCytomation) the localization of metallothionein and to determine MT expression in 9 patients with chronic pancreatitis. Our studies confirm that MT is present in exocrine and endocrine cells of patients with chronic pancreatitis and chronic pancreatitis with concomitant diabetes. They also indicate increased expression of MT, particularly in acinar cells of the pancreas. This suggests that MT is greatly involved in homeostasis of the pancreas and synthesis of pancreatic hormones.