HU Qingqing,ZHOU Enmin,Jin Ke, et al. Effect of melatonin on expression of α-SMA and E-cadherin in mice with bleomycin-induced pulmonary fibrosis[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2018, 48(8): 577-581.
Abstract：Objective: To investigate the effect of melatonin on expression of α-SMA and E-cadherin in mice with bleomycin induced pulmonary fibrosis and its mechanism. Methods: Mice pulmonary fibrosis model was made by intratracheal injection of bleomycin (5.0 mg/kg) and randomly divided as model group and melatonin group. After 4 weeks, the mice were sacrificed and lung tissue extracted. HE staining was used to evaluate the morphological changes and the oxidative stress kit was used to detect the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA). The levels of inflammatory cytokines IL-6 and TNF-α in lung tissues were measured by RT-PCR. The expression of α-SMA and E-cadherin in lung tissue was detected by immunohistochemistry. The expression of α-SMA and E-cadherin mRNA in lung tissue was detected by RT-PCR. Western blot was used to detect the expression of α-SMA and E-cadherin in lung tissue. Results: Compared with the model group, the SOD expression in the melatonin group was significantly increased (P<0.05), and the MDA expression was significantly decreased (P<0.05); the expression of TNF-α and IL-6 in the lung tissue was significantly decreased (P<0.05). Immunohistochemistry results showed that the expression of α-SMA protein in the lung tissue of melatonin group was significantly reduced (P<0.05), and E-cadherin protein expression was significantly increased (P<0.05); RT-PCR experiment results showed that the expression of α-SMA mRNA in the lungs of melatonin mice was significantly decreased (P<0.05), and E-cadherin mRNA expression was significantly increased (P<0.05). Western blot results showed that the expression of α-SMA protein in lung tissue of melatonin mice was significantly decreased (P<0.05), and E-cadherin protein expression was significantly increased (P<0.05). Conclusion: Melatonin can inhibit the expression of α-SMA protein, reduce the expression of E-cadherin, and delay the progression of pulmonary fibrosis by reducing the levels of oxidative stress and inflammatory factors.
[1] CHIOMA O S, DRAKE W P. Role of microbial agents in pulmonary fibrosis[J]. Yale J Biol Med, 2017, 90(2): 219-227.
[2] POVEDANO J M, MARTINEZ P, SERRANO R, et al. Therapeutic effects of telomerase in mice with pulmonary fibrosis induced by damage to the lungs and short telomeres [J]. Elife, 2018, 7. pii: e31299.
[3] REITER R J, TAN D X, ROSALES-CORRAL S, et al. The universal nature, unequal distribution and antioxidant functions of melatonin and its derivatives[J]. Mini Rev Med Chem, 2013, 13(3): 373-384.
[4] SHIN N R, PARK J W, LEE I C, et al. Melatonin suppresses fibrotic responses induced by cigarette smoke via downregulation of TGF-beta1[J]. Oncotarget, 2017, 8(56): 95692-95703.
[5] YEUNG H M, HUNG M W, LAU C F, et al. Cardioprotective effects of melatonin against myocardial injuries induced by chronic intermittent hypoxia in rats[J]. J Pineal Res, 2015, 58(1): 12-25.
[6] DAS N, MANDALA A, NAAZ S, et al. Melatonin protects against lipid-induced mitochondrial dysfunction in hepatocytes and inhibits stellate cell activation during hepatic fibrosis in mice[J]. J Pineal Res, 2017, 62: e12404.
[7] YOU X Y, XUE Q, FANG Y, et al. Preventive effects of Ecliptae Herba extract and its component, ecliptasaponin A, on bleomycin-induced pulmonary fibrosis in mice[J]. J Ethnopharmacol, 2015, 175: 172-180.
[8] MURTHA L A, SCHULIGA M J, MABOTUWANA N S,
et al. The processes and mechanisms of cardiac and pulmonary fibrosis[J]. Front Physiol, 2017, 8: 777.
[9] KAUR A, MATHAI S K, SCHWARTZ D A. Genetics in idiopathic pulmonary fibrosis pathogenesis, prognosis, and treatment[J]. Front Med (Lausanne), 2017, 4: 154.
[10] CHERESH P, KIM S J, TULASIRAM S, et al. Oxidative stress and pulmonary fibrosis[J]. Biochim Biophys Acta, 2013, 1832(7): 1028-1040.
[11] VAN LINTHOUT S, MITEVA K, TSCHOPE C. Crosstalk between fibroblasts and inflammatory cells[J]. Cardiovasc Res, 2014, 102(2): 258-269.
[12] LE T T, KARMOUTY-QUINTANA H, MELICOFF E, et al. Blockade of IL-6 Trans signaling attenuates pulmonary fi-brosis[J]. J Immunol, 2014, 193(7): 3755-3768.
[13] HOU J, MA T, CAO H, et al. TNF-alpha-induced NF-kappaB activation promotes myofibroblast differentiation of LR-MSCs and exacerbates bleomycin-induced pulmonary fibrosis[J]. J Cell Physiol, 2018, 233(3): 2409-2419.
[14] CHEN C, WANG Y Y, WANG Y X, et al. Gentiopicroside ameliorates bleomycin-induced pulmonary fibrosis in mice via inhibiting inflammatory and fibrotic process[J]. Biochem Biophys Res Commun, 2018, 495(4): 2396-2403.
[15] STEHLE J H, SAADE A, RAWASHDEH O, et al. A survey of molecular details in the human pineal gland in the light of phylogeny, structure, function and chronobiological diseases [J]. J Pineal Res, 2011, 51(1): 17-43.
[16] WANG M L, WEI C H, WANG W D, et al. Melatonin attenuates lung ischaemia-reperfusion injury via inhibition of oxidative stress and inflammation[J]. Interact Cardiovasc Thorac Surg, 2018, 26(5): 761-767.
[17] ZHAO H, WU Q Q, CAO L F, et al. Melatonin inhibits endoplasmic reticulum stress and epithelial-mesenchymal transition during bleomycin-induced pulmonary fibrosis in mice[J]. PLoS One, 2014, 9(5): e97266.
[18] KARIMFAR M H, ROSTAMI S, HAGHANI K, et al. Melatonin alleviates bleomycin-induced pulmonary fibrosis in mice[J]. J Biol Regul Homeost Agents, 2015, 29(2): 327-334.
[19] VERES-SZEKELY A, PAP D, SZIKSZ E, et al. Selective measurement of alpha smooth muscle actin: why beta-actin can not be used as a housekeeping gene when tissue fibrosis occurs[J]. BMC Mol Biol, 2017, 18(1): 12.
[20] SUNILGOWDA S N, NAGARAJAN D. Radiation-Induced Pulmonary epithelial-mesenchymal transition: a review of targeting molecular pathways and mediators[J]. Curr Drug Targets, 2018, 19(10): 1191-1204.
[21] TASHIRO J, RUBIO G A, LIMPER A H, et al. Exploring animal models that resemble idiopathic pulmonary fibrosis [J]. Front Med (Lausanne), 2017, 4: 118.
