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Ghada M.A. Mazen Biological Applications Department, Nuclear Research Center, Atomic Energy Authority, Egypt. For correspondence Email [email protected] Tel 202-155-0540340 Non-Pharmaceutical Cures for Acute Pancreatitis Induced by Cerulein in Rats Abstract Conclusion These results showed that NAC possesses significant mitigating in all studied parameters. Whereas, incorporation of NAC and ALA induced preeminent effects and can be represent as an ideal cures for acute pancreatitis. There are still no specific therapies for AP but the opinion that oxidative stress is one of the precursors of acute pancreatitis merited our attention to address the possible protective effects of one of the nutraceuticals known to have the ability to resist oxidation, namely N-acetylcysteine. NAC, a thiol sulfhydryl containing compound derived from the sulfur-containing amino acid cysteine. It is a precursor of intracellular glutathione (GSH) and thus a ROS scavenger 4. The anti-inflammatory effects of NAC have also been confirmed through observation of nuclear factor kappa B (NF-kB) inhibition 5 and reduce the expression of cytokines and chemokines as well as the pancreatic damage 6. Alpha-lipoic acid (ALA) is natural compound widely distributed in plants and animals. It is a strong antioxidant with anti-inflammatory effect due to its property of both forms oxidized (LA) and reduced (dihydrolipoic acid DHLA). Thus, ALA is called a universal antioxidant 7.A significant activity of LA and its derivatives are the capability to scavenger free radical and to chelate toxic metals as well as to increase glutathione content inside the cells 8. Experimental animals Healthy 40 adult male albino rats (150-180g) were used in the current study and obtained from the Animal House of the Nuclear Research Center, Atomic Energy Authority, Cairo, Egypt. The animals were maintained in the animal-holding room under normal conditions (12/12 h light/dark cycle, 50-70 humidity and 30C). They were fed on standard laboratory diet and water ad libitum. All rat procedures were carried out in accordance with the Ethics Committee of the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH publication No. 85-23, 1996). Animal groups and experimental protocol The rats were randomly assigned to four groups each contain ten rats and treated as follows Control group Rats received daily subcutaneous injection of normal saline (0.9NaCl) for three days. At the completion of each experimental period (24wks), the animals were anaesthetized and blood samples were collected by cardiac puncture. Sera were separated and divided into considerable aliquots to avoid the effects of repeated thawing and freezing. Preparation of pancreatic homogenates Pancreatic tissue (0.1-0.2g) was homogenized in 2ml of 0.2M Tris-HCl (pH 8.0) containing 0.05M CaCl2 in a tissue mixer. DNA, RNA and proteins were extracted from HClO4 precipitates of an aliquot of the homogenate. DNA and RNA content were measured by the method of Burton 9 and Ceriotti 10 respectively. The protein level was determined by the method of Bradford 11. Biochemical analysis RESULTS Effect of cerulein on tumor markers Table (4) shows the both body and pancreatic weight in cerulein rats group were significantly (p0.05) decreased. Moreover, DNA, RNA and protein level were significantly (p0.05) decline as compared to the control group. Conversely, treatment by NAC alone or together with ALA led to a remarkable improvement in the body and pancreatic weight as well as a considerable correction in the levels of DNA, RNA and protein dependent on the time of treatment (24 wks). Table 1 Effect of NAC alone or plus ALA on tumor markers in pancreatic rats induced by cerulein. GroupsAcute Pancreatitis RatsControlRecoveryNACNACALACEA (ng/ml)2 wks0.18 0.004 Da2.54 0.040 A a1.35 0.045 Ba1.02 0.049 C a4 wks0.19 0.004 D b3.21 0.063 A b1.01 0.044 Bb0.79 0.031 C bCA242 (U/L)2 wks 4 wks107.95 1.01 Da 109.94 1.12 Db425.39 7.43 Aa 390.58 6.88 Ab356.75 5.67 Ba 259.31 4.35 Bb238.89 5.86 Ca 148.32 1.512 CbTAP (ng/ml)2 wks 4 wks10.15 0.269 Da 9.96 0.259 Db26.85 0.929 Aa 30.88 0.991 Ab19.31 0.794 Ba 15.13 0.417 Bb16.12 0.603 Ca 11.78 0.821 Cb- A, B, C, D Means with different superscript within a row are significantly different at (P0.05). – a,b Means with different subscript between two points (24 wks) for the same parameters are significantly different at (P0.05). Table 4 Effect of NAC alone or plus ALA on some physiological parameters in pancreatic rats induced by cerulein. GroupsControlAcute Pancreatitis RatsRecoveryNACNACALABody weight (g)2 wks186.149.387 Aa 141.586.939 Da 153.977.174 Ca175.338.625 Ba 4 wks217.4811.541 Ab172.028.953 Db192.3610.251 Cb210.1411.263 BbPancreatic weight (g)2 wks0.94 0.011 Aa0.76 0.079 Da0.79 0.074 Ca0.86 0.069 Ba4 wks1.22 0.0296 Ab0.81 0.0751 Db0.85 0.066 Cb1.07 0.061 BbDNA (mg)2 wks1.163 0.021 Aa1.047 0.019 Da1.082 0.023 Ca1.155 0.026 Ba4 wks1.318 0.026 Ab1.088 0.022 Db1.117 0.026 Cb1.276 0.028 BbRNA (mg)2 wks5.612 0.074 Aa4.139 0.048 Da4.707 0.052 Ca5.258 0.063 Ba4 wks5.994 0.079 Ab4.627 0.054 Db5.101 0.061 Cb5.791 0.075 BbProtein (mg/g tissue)2 wks64.361 2.649 Aa51.921 2.012 Da55.647 2.086 Ca60.813 2.227 Ba4 wks69.027 3.004 Ab54.374 2.057 Db60.553 2.115 Cb67.616 2.794 Bb- A, B, C, D Means with different superscript within a row are significantly different at (P0.05). – a,b Means with different subscript between two points (24 wks) for the same parameters are significantly different at (P0.05). DISCUSSION AP is a complicated disorder whose detrimental effects may surpass the pancreas to multiorgan failure. The release of reactive oxygen species (ROS) and inflammatory cytokines within the pancreas are known to induce the evolution of pancreatitis with varying degrees of certainty 12. Cerulein-induced pancreatitis in animal model has been considered the most relevant model that resembles the human phenotype in many aspects. In the current work, a significant elevation occurred in the tumor marker (CEA CA242) as a result of cerulein administration in rats (Table 1). These results may be due to defect in the immune system defense or the increment of free radical production and lipid peroxidation concentration. Several authors suggested that the elevation in the activities of phospholipase A2 and cyclooxygenase-2 may lead to pronounce acute pancreatitis in human 13 and in experimental animals 14. The obtained data showed that pancreatitis rats recorded a considerable depletion in the both of body and pancreatic weights associated with a significant decline in the level of total protein and concentrations of DNA RNA. These data are in harmony with the results obtained by Sandeep Veeresh 19. Acinar cells are the protein factory of the body, consequently, in AP catabolism of proteins could increase up to 80 thus, a sharp decline in protein content was observed in pancreas 20. Moreover, free radicals when left unchecked wreak havoc on cells causing damage to membranes and DNA. Then, these free radicals have the potential for such devastating damage, considered to be a major factor in the cancer and aging processes 21. Although extensive researches in the field, there is no specific therapy against AP and treatment is mainly limited to supportive care. There exists a positive feedback between oxidative stress and cytokines which represent the most critical factors in the outcome of AP. Taken together, the main objective of the current study was to address whether NAC alone or in combination with ALA could be considered an appropriate treatment against acute pancreatitis. Y, dXiJ(x(I_TS1EZBmU/xYy5g/GMGeD3Vqq8K)fw9
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