Content of NCAM in the brain and pancreas of rats in response to endointoxication under conditions of experimental chronic pancreatitis

  • V. A. Makarchuk SI «Institute of Gastroenterology of the National Academy of Medical Sciences of Ukraine», Dnipropetrovsk
  • G. А. Ushakova Oles Honchar Dnipropetrovsk National University
Keywords: molecules with average weight, TBA-active products, chronic pancreatitis, brain, pancreas, neuronal cell adhesion molecule


The study was undertaken to examine the influence of chronic pancreatitis on the distribution of neuronal cell adhesion molecule (NCAM) in the pancreas and various brain regions of rats under the conditions of endogenous intoxication. The study was conducted using 36 white nonlinear male rats (6 months old, 190–220 g). To develop the state of chronic pancreatitis, animals were subjected tolaparotomy under general anesthesia and prolonged occlusion of the pancreatic duct. The morphological examination of pancreatic tissue hasbeen performed to confirm the chronic pancreatitis development in animals. Biochemical evaluation of the pancreatic fibrosis has been performed by measuring plasma levels of hyaluronic acid, hydroxyproline and protein-free hydroxyproline. The intensity of free radical oxidation has been assessed by the change in the concentration of TBA-active products in plasma. The level of endotoxemia has been determinedby the content of average weight molecules in plasma. Protein fractions were extracted from the pancreas and various parts of the rat brain and the levels of soluble (sNCAM) and membrane (mNCAM) proteins were studied with the use of the competitive ELISA. Total protein in the obtained fractions was measured by the Bradford assay. Occlusion of the pancreatic duct resultedin significant atrophy of acinar tissue, fibrosis and disfunction of the pancreas along with the decreasing in the antioxidant defense of animals. The present study shows developing of endointoxication in experimentalrats, signified by considerable increase of molecules with average weight in plasma due to the activation of lipid peroxidation. It was established that, as a result of the experimental pancreas dysfunction, significant redistribution of soluble and membrane forms of NCAM took place, more especially in the cerebellum and thalamus of rats; it caused changing of cell-cell adhesion in these brain regions. Multidirectional NCAM distribution in the cerebellum and thalamus gives evidence of neuroplasticity as specific adaptation under conditions of chronic pancreatitis.


Alfyorova, V.V., Uzbekov, M.G., Misionzhnik, E.Y., Loukyanyuk, E.V., Geht, A.B., Shklovsky, V.M., Shikhov, S.N., 2011. Serologicheskie markeri endogennoj intoksikacii v kompleksnoj ocenke reabilitacionnogo potenciala bol'nih, perenesshih ishemicheskij insyl't [Serological markers of endogeneous intoxication in determining rehabilitation potential for the patients after ischemic stroke]. Soc. Klin. Psihiatriia 11(3), 54–57 (in Russian).

Bisaz, R., Conboy, L., Sandi, C., 2009. Learning under stress: A role for the neural cell adhesion molecule NCAM. Neurobiol. Learn. Mem. 91, 333–342. >> doi.:10.1016/j.nlm.2008.11.003

Bisaz, R., Schachner, M., Sandi, C., 2011. Causal evidence for the involvement of the neural cell adhesion molecule, NCAM, in chronic stress-induced cognitive impairments. Hippocampus 21, 56–71. >> doi.:0.1002/hipo.20723

Bradford, M., 1985. Rapid and sensitive methods for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.

Campodonico, P.B., Bal de Kier Joffe, E.D., Urtreger, A.J., Lauria, L.S., Lastiri, J.M., Puricelli, L.I., Todaro, L.B., 2010. The neural cell adhesion molecule is involved in the metastatic capacity in a murine model of lung cancer. Mol. Carcinog. 49, 386–397. >> doi.:10.1002/mc.20611

Catalá, A., 2009. Lipid peroxidation of membrane phospholipids generates hydroxy-alkenals and oxidized phospholipids active in physiological and/or pathological conditions. Chem. Phys. Lipids 157, 1–11. >> doi.:10.1016/j.chemphyslip.2008.09.004

Çöl, C., Dinler, K., Hasdemir, O., Büyükaşik, O., Buğdayci, G., 2010. Oxidative stress and lipid peroxidation products: Effect of pinealectomy or exogenous melatonin injections on biomarkers of tissue damage during acute pancreatitis. Hepatobiliary Pancreat. Dis. Int. 9(1), 78–82.

Ding, Z., Liu, J., Lin, R., Hou, X.H., 2012. Experimental pancreatitis results in increased blood-brain barrier permeability in rats: A potential role of MCP-1. J. Dig. Dis. 13, 179–185. >> doi.: 10.1111/j.1751-2980.2011.00568.x

Ďurackova, Z., 2010. Some current insights into oxidative stress. Physiol. Res. 59, 459–469.

Duterme, C., Mertens-Strijthagen, J., Tammi, M., Flamion, B., 2009. Two novel functions of hyaluronidase-2 (Hyal2) are formation of the glycocalyx and control of CD44-ERM interactions. J. Biol. Chem. 284(48), 33495–33508. >> doi.:10.1074/jbc.M109.044362

Eleeva, M.A-K., 2013. Sostojanie prooksidantnoj sistemi krovi i uroven' molekul srednej massi u bol'nyh s insul'tom [The condition of prooxidant blood system and medium mass molecules level in the patients under insult]. Vestnik Novih Med. Tehnologij (Elektronnij Zh.) 1 (in Russian).

El-Shabrawi Mortada, H.F., Zein El Abedin, M.Y., Omar, N., Kamal, N.M., Elmakarem, A.S., Khattab, S., El-Sayed, H.M., El-Hennawy, A., Ali Ali, S.M., 2012. Predictive accuracy of serum hyaluronic acid as a non-invasive marker of fibrosis in a cohort of multi-transfused Egyptian children with b-thalassaemia major. Arab J. Gastroenterol. 13, 45–48. >> doi.:10.1016/j.ajg.2012.06.006

Fogel, A.I., Stagi, M., Arce, K.P., Biederer, T., 2011. Lateral assembly of the immunoglobulin protein SynCAM 1 controls its adhesive function and instructs synapse formation. EMBO J. 30(23), 4728–4738. >> doi.Ж10.1038/emboj.2011.336

Gold, E.W., 1981. The quantitative spectrophotometric estimation of total sulfated glycosaminoglican levels. Biochem. Biophys. Acta 673, 408–415. >> doi.:10.1016/0304-4165(81)90472-4

Gomazkov, O.A., 2011. Starenie mozga i nejrotroficheskaja terapija [The aging of the brain and neurotrophic therapy]. Izdatel'stvo ICAR, Moscow (in Russian).

Grigsby, B., Rodriguez, H., Khan, K., 2012. Antioxidants and chronic pancreatitis: Theory of oxidative stress and trials of antioxidant therapy. Dig. Dis. Sci. 57, 835–841. >> doi.:10.1007/s10620-012-2037-3

Gubergrits, N.B., 2013. Sovremennye dostizhenija evropejskoj pancreatologii [Modern achievements of european pancreatology]. Gastroenterologija 47(1), 65–71 (in Russian).

Hagiyama, M., Ichiyanagi, N., Kimura, K.B., Murakami, Y., Ito, A., 2009. Expression of a soluble isoform of cell adhesion molecule 1 in the brain and its involvement in directional neurite outgrowth. Am. J. Pathol. 174(6), 2278–2289. >> doi.:10.2353/ajpath.2009.080743

Hinkle, C.L., Diestel, S., Lieberman, J., Maness, P.F., 2006. Metalloprotease-induced ectodomain shedding of neural cell adhesion molecule (NCAM). J. Neurobiol. 66, 1378–1395. >> doi.:10.1002/neu.20257

Hübschmann, M.V., Skladchikova, G., 2010. The role of ATP in the regulation of NCAM function. Ed. V. Berezin. Springer Science + Business Media, LLC, 663, pp. 81–89.

Kleene, R., Cassens, C., Bahring, R., Theis, T., Xiao, M.-F., Dityatev, A., Schafer-Nielsen, C., Doring, F., Wischmeyer, E., Schachner, M., 2010. Functional consequences of the interactions among the neural cell adhesion molecule NCAM, the receptor tyrosine kinase TrkB, and the inwardly rectifying K+ channel KIR3.3. J. Biol. Chem. 285(37), 28968–28979. >> doi.:10.1074/jbc.M110.114876

Kleene, R., Mzoughi, M., Joshi, G., Kalus, I., Bormann, U., Schulze, C., Xiao, M.-F., Dityatev, A., Schachner, M., 2010. NCAM-induced neurite outgrowth depends on binding of calmodulin to NCAM and on nuclear import of NCAM and fak fragments. J. Neurosci. 32(30), 10784–10798. >> doi.:10.1523/JNEUROSCI.0297-10.2010

Krotenko, N.M., Boiko, A.S., Yepanchintseva, Y.M., Ivanova, S.A., 2012. Pokazateli okislitel'nogo stressa i endogennoj intoksikacii v perifericheskoj krovi u bol'nih s ekzogenno-organicheskimi rasstrojstvami v dinamike farmakoterapii [Parameters oxidative stress and endogenous intoxication of peripheral blood in patients with exogenous organic disorders in dynamic of the pharmacotherapy]. Bjulleten' Sibirskoj Medicini 1, 179–185 (in Russian).

Loury, O.H., Rosenbrough, N.J., Farr, R.L., Randall, R.J., 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193(2), 265–275.

Madro, A., Slomka, M., Celinski, K., 2011. Can we expect progress in the treatment of fibrosis in the course of chronic pancreatitis? Adv. Med. Sci. 56, 132–137. >> doi.:10.2478/v10039-011-0023-1

Makarchuk, V.A., Ushakova, G.A., Krylova, O.O., 2013. Stan sistemi glutatіonu in krovі schurіv ta osoblivostі morfologіchnih zmin tkanini pіdshlunkovoi zalozi za umov eksperimentalnogo gostrogo ta hronichnogo pancreatitu [The state of glutathione system in the rat blood and peculiarities of morphological changes of the pancreas under experimental acute and chronic pancreatitis]. Ukr. Biohim. Zh. 85(1), 45–52 (in Ukrainian).

Mao, X., Schwend, T., Conrad, G.W., 2012. Expression and localization of neural cell adhesion molecule and polysialic acid during chick corneal development. Invest. Ophthalmol. Vis. Sci. 53(3), 1234–1243. >> doi.:10.1167/iovs.11-8834

Masamune, A., Watanabe, Т., Kikuta, K., Shimosegawa, T., 2009. Roles of pancreatic stellate cells in pancreatic Inflammation and fibrosis. Clin. Gastroenterol. Hepatol. 7, 48–54. >>doi.:10.1016/j.cgh.2009.07.038

Merinova, N.I., Kozlova, N.M., Kolsnichenko, L.S., Suslov, A.I., Leonov, Z.A., 2013. Pokazateli perekisnogo okislenija lipidov i glutationovoj antioksidantnoj zasshiti u bol'nih s obostreniem chronicheskogo pankreatita [Indicators of lipid peroxidation and glutathione antioxidant defense in patients with exacerbation of chronic pancreatitis]. Saratov J. Med. Sci. Res. 9(2), 259–262 (in Russian).

Ngo, T.T., Lenhoff, H.M., Yaklich, A., 1988. Immunofermentnij analiz [Enzyme immunoassay]. Ed. T.T. Ngo, G. Lenhoff. Mir, Moscow (in Russian).

Nielsen, J., Gotfryd, K., Li, S., Kulahin, N., Soroka, V., Rasmussen, K.K., Bock, E., Berezin, V., 2009. Role of glial cell line-derived neurotrophic factor (GDNF)–neural cell adhesion molecule (NCAM) interactions in induction of neurite outgrowth and identification of a binding site for NCAM in the heel region of GDNF. J. Neurosci. 36(29), 11360–11376. >> doi.Ж10.1523/JNEUROSCI.3239-09.2009

Nikolskaya, V., Danilchenko, U., Memetova, Z., 2013. Biohimicheskij aspekt rassmotrenija roli molekul srednej massi v organizme [Biochemical aspects of the consideration of the role of high molecular mass in the body]. Uchenie Zapiski Tavricheskogo Nac. Univ. im. V.I. Vernadskogo Serija Biologija, Himija 26(1), 139–145 (in Russian).

Nykolaychik, V.V., Moyn, V.M., Kirkovskiy, V. Mazur, L.I., Lobacheva, G.A., Bychko, T.N., 1991. Sposob opredelenija «Srednih molekul» [Method for determining «average molecules»]. Lab. Delo 10, 13–18 (in Russian).

Osadchuk, M.A., 1979. Metodi issledovanija oksiprolina v krovi i moche [Methods of hydroxyproline in blood and urine]. Lab. Delo 8, 456–458 (in Russian).

Ovsyannikova, L.M., Alekhin, S.M., Drobinska, A., 1999. Biochimichni i biofizychni metodi ocinky porushen' okisljuval'nogo gomeostazu v osib, scho zaznali radiacijnogo vplivu vnaslidok avarii na ChAES [Biochemical and biophysical methods for assessing violations of oxidative homeostasis in individuals exposed to radiation exposure due to the Chernobyl accident]. Chornobilinterinform, Kyiv (in Ukrainian).

Page, B.J., Toit, D.F., Muller, C.J.F., Mattysen, J., Lyners, R., 2000. An immunocytochemical profile of the endocrine pancreas using an occlusive duct ligation model. Journal of the Pancreas 4(1), 191–203.

Petri, A., Sabin, K., 2003. Nagljadnaja statistika v medicine [Visual statistics in medicine]. Geotar-Med, Moscow (in Russian).

Sapozhnikov, A.G., Dorosevich, A.E., 2000. Histologicheskaja i mikroskopicheskaja tehnika [The histological and microscopic technique], Izdatel'stvo SAU, Smolensk (in Russian).

Shklovsky, V.M., Alfyorova, V.V., Misionzhnik, E.Y., Krasnov, V.N., Gusev, E.I., Uzbekov, M.G., Gekht, A.B., Lukyanyuk, E.V., 2011. Znachenie sindroma endogennoj intoksikacii dlja vosstanovlenija narushennih funkcij posle ishemicheskogo insul'ta [The role of autointoxication in functional rehabilitation after ischemic stroke]. Nevrologija 63(4), 27–30 (in Russian).

Tereshenko, O.A., Botashev, A.A., Pomeshik, Y.V., Petrosyan, E.A., Sergienko, V.I., 2012. Sindrom endogennoj intoksikacii i sistemnoj vospalitel'noj reakcii pri zhelchnom peritonite, oslozhnennom abdominal'nim sepsisom [Endogenous intoxication syndrome and system inflammatory reaction in choleperitonitis, complicated by abdominal sepsis]. Vestnik Eksp. i Klin. Hirurgii 5(4), 722–726 (in Russian).

Tropina, I.V., Morozov, S.V., Dolgih, V.T., Poluektov, V.L., Ershov, A.V., 2010. Endogennaja intoksikacija u bol'nih s postnekroticheskimi kistami podzheludochnoj zhelezi [Endogenous intoxication in patients with postnecrotic pancreatic cysts]. Vestnik VolGMY 33(1), 97–98 (in Russian).

Uzbekov, M.G., Alferova, V.V., Gekht, A.B., Misionzhnik, E.Y., 2010. K voprosu o patohimicheskih izmenenijah sistemi metabolicheskogo gomeostaza v ostrom periode ishemicheskogo insyl'ta [About pathochemical changes of the system of metabolic homeostasis in acute period of ischemic stroke]. Ross. Mediko-Biologicheskij Vestnik im. I.P. Pavlova 3, 72–76 (in Russian).

Venkatasubramanian, P.N., 2012. Imaging the pancreatic ECM. Ed. P.J. Grippo, H.G. Munshi. Transworld Research Network, pp. 11–27.

Zecchini, S., Bombardelli, L., Decio, A., Bianchi, M., Mazzarol, G., Sanguineti, F., Aletti, G., Maddaluno, L., Berezin, V., Bock, E., Casadio, C., Viale, G., Colombo, N., Giavazzi, R., Cavallaro, U., 2011. The adhesion molecule NCAM promotes ovarian cancer progression via FGFR signaling. EMBO Mol. Med. 3(8), 480–494. >> doi.:10.1002/emmm.201100152

Zhang, X.-P., Hua, T., 2007. Pathogenesis of pancreatic encephalopathy in severe acute pancreatitis. Hepatobiliary Pancreat. Dis. Int. 6(2), 134–140.


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