Ying HE, Shengzhong LIU, Weifeng LU, Keli HUANG, Tao YU

Osthol sıçanları miyokardiyal iskemi/reperfüzyona karşı korumak için miR-30a’yı azaltarak düzenler ve otofajiyi destekler

Amaç: Bu çalışmada bir miyokardiyal iskemi/reperfüzyon hasarı modelinde osthol tedavisi sonrası miR-30a ekspresyonu ve miyokardiyal otofajideki değişiklikler araştırıldı. Çalışma planı: Otuz erkek Wistar sıçanı (ağırlık, 170-220 g, yaş 8 hafta) randomize şekilde kontrol (taklit) grubu, iskemi/ reperfüzyon (model) grubu ve iskemi/reperfüzyon + osthol tedavi sonrası (osthol) grubu olmak üzere üç gruba ayrıldı. Miyokardiyal kollajen değişiklilerini saptamak için Masson'un trikrom boyaması kullanıldı. İskemik alandaki apoptotik kardiyomiyositler terminal deoksinükleotidil transferaz aracılı dUTP (2'-deoxyuridine 5'-triphosphate) çentik-uç etiketleme testi ile in situ etiketlendi. Miyokardiyal dokuda LC3b (light chain 3 beta) ve Beclin-1 otofaji belirteçlerinin düzeyleri western blotlama ile saptandı. miR-30a ekspresyonu kantitatif ters transkripsiyon polimeraz zincir reaksiyonu ile saptandı. Bulgular: İskemi/reperfüzyon, taklit grubuna kıyasla kollajen içeriklerini anlamlı şekilde artırdı. Osthol, iskemi/reperfüzyonun artırdığı kollajen içeriklerini anlamlı şekilde engelledi. Osthol, iskemi/reperfüzyonun artırdığı miyokardiyal fibrozu, miyokardiyal şişkinliği, nekrozu ve miyokardiyal atrofiyi engelledi. Osthol, miyokardiyal hücrelerde iskemi/reperfüzyonun artırdığı apoptozu da anlamlı şekilde engelledi. Ayrıca, LC3b-I’in LC3b-II’ye dönüşümü ve Beclin-1 ekspresyonu iskemi/reperfüzyon tarafından anlamlı şekilde engellendi. Osthol tedavisi iskemi/reperfüzyon sıçanlarında LC3b-I’in LC3b-II’ye dönüşümünü ve Beclin-1 ekspresyonunu anlamlı şekilde artırdı. Son olarak, miR-30a ekspresyonu iskemi/reperfüzyon sıçanlarında anlamlı şekilde artarken osthol, miR-30a ekspresyonunu baskıladı. Sonuç: Osthol otofajiyi destekleyerek miyokardiyal iskemi/ reperfüzyon hasarını azalttı. Osthol miR-30a ekspresyonunu azaltarak düzenleyerek otofaji sırasında miyokardı korur.

Osthole down-regulates miR-30a and promotes autophagy to protect rats against myocardial ischemia/reperfusion injury

Background: This study aims to investigate the changes inmiR-30a expression and myocardial autophagy following ostholetreatment in a myocardial ischemia/reperfusion injury model.Methods: Thirty male Wistar rats (weighing 170 to 220 g, aged8 weeks) were randomly divided into three groups as control(sham) group, ischemia/reperfusion (model) group, and ischemia/reperfusion + osthole post-treatment (osthole) group. Masson’strichrome staining was used to detect myocardial collagen changes.Apoptotic cardiomyocytes in the ischemic area were labeledin situ by terminal deoxynucleotidyl transferase-mediated dUTP(2'-deoxyuridine 5'-triphosphate) nick end labeling assay. Levelsof autophagy markers light chain 3 beta (LC3b) and Beclin-1 inmyocardial tissue were detected by western blotting. Expressionof miR-30a was detected by quantitative reverse transcriptionpolymerasechain reaction.Results: Compared with the sham group, ischemia/reperfusionsignificantly increased collagen contents. Osthole significantlyinhibited the ischemia/reperfusion-increased collagen contents.Osthole inhibited the ischemia/reperfusion-increased myocardialfibrosis, myocardial swelling, necrosis, and myocardial atrophy.Osthole also significantly inhibited the ischemia/reperfusionincreasedapoptosis of myocardial cells. Moreover, the conversionof LC3b-I to LC3b-II and the Beclin-1 expression were significantlyinhibited by ischemia/reperfusion. Osthole treatment significantlyincreased the conversion of LC3b-I to LC3b-II and Beclin-1expression in ischemia/reperfusion rats. Finally, the expression ofmiR-30a was significantly increased in ischemia/reperfusion rats,while Osthole suppressed the expression of miR-30a.Conclusion: Osthole promoted autophagy, thereby alleviatingmyocardial ischemia/reperfusion injury. Osthole protects themyocardium during autophagy by down-regulating miR-30a expression.

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1. Sciarretta S, Volpe M, Sadoshima J. Mammalian target of rapamycin signaling in cardiac physiology and disease. Circ Res 2014;114:549-64.
2. Ouyang C, You J, Xie Z. The interplay between autophagy and apoptosis in the diabetic heart. J Mol Cell Cardiol 2014;71:71-80.
3. Zeng Y. Advances in mechanism and treatment strategy of cancer. Cell Mol Biol (Noisy-le-grand) 2018;64:1-3.
4. Ma X, Godar RJ, Liu H, Diwan A. Enhancing lysosome biogenesis attenuates BNIP3-induced cardiomyocyte death. Autophagy 2012;8:297-309.
5. Fukui M, Yamabe N, Choi HJ, Polireddy K, Chen Q, Zhu BT. Mechanism of Ascorbate-Induced Cell Death in Human Pancreatic Cancer Cells: Role of Bcl-2, Beclin 1 and Autophagy. Planta Med 2015;81:838-46.
6. Vacek TP, Vacek JC, Tyagi SC. Mitochondrial mitophagic mechanisms of myocardial matrix metabolism and remodelling. Arch Physiol Biochem 2012;118:31-42.
7. Petrovski G, Das S, Juhasz B, Kertesz A, Tosaki A, Das DK. Cardioprotection by endoplasmic reticulum stress-induced autophagy. Antioxid Redox Signal 2011;14:2191-200.
8. Wei C, Hu B, Shen E. Pivotal role of microRNAs in cardiac development and heart diseases. Chin J Pathophysiol 2011;27: 611-5.
9. Zhang ZR, Leung WN, Cheung HY, Chan CW. Osthole: A review on its bioactivities, pharmacological properties, and potential as alternative medicine. Evid Based Complement Alternat Med 2015;2015:919616.
10. Duan J, Yang Y, Liu H, Dou PC, Tan SY. Osthole ameliorates acute myocardial infarction in rats by decreasing the expression of inflammatory-related cytokines, diminishing MMP-2 expression and activating p-ERK. Int J Mol Med 2016;37:207-16.
11. Zhang Z, Pan C, Wang HZ, Li YX. Protective effects of osthole on intestinal ischemia-reperfusion injury in mice. Exp Clin Transplant 2014;12:246-52.
12. Wang XY, Dong WP, Bi SH, Pan ZG, Yu H, Wang XW, et al. Protective effects of osthole against myocardial ischemia/ reperfusion injury in rats. Int J Mol Med 2013;32:365-72.
13. Wu J, Yang Y, Xun N, Zeng L, Li Z, Yang W, et al. Osthole attenuates myocardial ischemia/reperfusion injury in rats by inhibiting apoptosis and inflammation. Am J Transl Res 2018;10:1109-16.
14. Peng C, Rao W, Zhang L, Gao F, Hui H, Wang K, et al. Mitofusin 2 Exerts a Protective Role in Ischemia Reperfusion Injury Through Increasing Autophagy. Cell Physiol Biochem 2018;46:2311-24.
15. Ma X, Liu H, Foyil SR, Godar RJ, Weinheimer CJ, Diwan A. Autophagy is impaired in cardiac ischemia-reperfusion injury. Autophagy 2012;8:1394-6.
16. Ohsumi Y. Molecular dissection of autophagy: two ubiquitinlike systems. Nat Rev Mol Cell Biol 2001;2:211-6.
17. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 2000;19:5720-8.
18. Vander Heide RS, Steenbergen C. Cardioprotection and myocardial reperfusion: pitfalls to clinical application. Circ Res 2013;113:464-77.
19. Kang R, Zeh HJ, Lotze MT, Tang D. The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ 2011;18:571-80.
20. Cheng Y, Zhu P, Yang J, Liu X, Dong S, Wang X, et al. Ischaemic preconditioning-regulated miR-21 protects heart against ischaemia/reperfusion injury via anti-apoptosis through its target PDCD4. Cardiovasc Res 2010;87:431-9.
21. Wazir R, Luo DY, Dai Y, Yue X, Tian Y, Wang KJ. Expression and proliferation profiles of PKC, JNK and p38MAPK in physiologically stretched human bladder smooth muscle cells. Biochem Biophys Res Commun 2013;438:479-82.
22. Zhu H, Wu H, Liu X, Li B, Chen Y, Ren X, et al. Regulation of autophagy by a beclin 1-targeted microRNA, miR-30a, in cancer cells. Autophagy 2009;5:816-23.