Mst1 silencing alleviates hypertensive myocardial injury associated with the augmentation of microvascular endothelial cell autophagy

Wang,Ling-Peng; Han,Rui-Mei; Wu,Bin; Luo,Meng-Yao; Deng,Yun-Hui; Wang,Wei; Huang,Chao; Xie,Xiang; Luo,Jian

doi:10.3892/ijmm.2022.5202

Mst1 silencing alleviates hypertensive myocardial injury associated with the augmentation of microvascular endothelial cell autophagy

Authors:
- Ling-Peng Wang
- Rui-Mei Han
- Bin Wu
- Meng-Yao Luo
- Yun-Hui Deng
- Wei Wang
- Chao Huang
- Xiang Xie
- Jian Luo
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Affiliations: Department of Cardiology, The First Affiliated Hospital, Xinjihypang Medical University, Urumqi, Xinjiang 830000, P.R. China, Department of Cardiology, Shanghai Xuhui Central Hospital, Shanghai 200031, P.R. China, Department of Geriatrics, Xinjiang Military General Hospital, Urumqi, Xinjiang 830000, P.R. China, Department of Internal Medicine, The First Affiliated Hospital, Xinjiang Medical University, Urumqi, Xinjiang 830000, P.R. China
Published online on: November 9, 2022 https://doi.org/10.3892/ijmm.2022.5202
Article Number: 146
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The activation of mammalian ste20‑like kinase1 (Mst1) is a crucial event in cardiac disease development. The inhibition of Mst1 has been recently suggested as a potential therapeutic strategy for the treatment of diabetic cardiomyopathy. However, whether silencing Mst1 also protects against hypertensive (HP) myocardial injury, or the mechanisms through which this protection is conferred are not yet fully understood. The present study aimed to explore the role of Mst1 in HP myocardial injury using in vivo and in vitro hypertension (HP) models. Angiotensin II (Ang II) was used to establish HP mouse and cardiac microvascular endothelial cell (CMEC) models. CRISPR/adenovirus vector transfection was used to silence Mst1 in these models. Using echocardiography, hematoxylin and eosin staining, Masson's trichrome staining, the enzyme‑linked immunosorbent assay detection of inflammatory factors, the enzyme immunoassay detection of oxidative stress markers, terminal deoxynucleotidyl transferase dUTP nick‑end labeling staining, scanning electron microscopy, transmission electron microscopy, as well as immunofluorescence and western blot analysis of the autophagy markers, p62, microtubule‑associated proteins 1A/1B light chain 3B and Beclin‑1, it was found that Ang II induced HP myocardial injury with impaired cardiac function, increased the expression of inflammatory factors, and elevated oxidative stress in mice. In addition, it was found that Ang II reduced autophagy, enhanced apoptosis, and disrupted endothelial integrity and mitochondrial membrane potential in cultured CMECs. The silencing of Mst1 in both in vivo and in vitro HP models attenuated the HP myocardial injury. On the whole, these findings suggest that Mst1 is a key contributor to HP myocardial injury through the regulation of cardiomyocyte autophagy.

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1	Oparil S, Acelajado MC, Bakris GL, Berlowitz DR, Cífková R, Dominiczak AF, Grassi G, Jordan J, Poulter NR, Rodgers A and Whelton PK: Hypertension. Nat Rev Dis Primers. 4:180142018. View Article : Google Scholar : PubMed/NCBI
2	McCormack T, Krause T and O'Flynn N: Management of hypertension in adults in primary care: NICE guideline. Br J Gen Pract. 62:163–164. 2012. View Article : Google Scholar : PubMed/NCBI
3	Gonzalez A, Ravassa S, Lopez B, Moreno MU, Beaumont J, José GS, Querejeta R, Bayés-Genís A and Díez J: Myocardial remodeling in hypertension. Hypertension. 72:549–558. 2018. View Article : Google Scholar : PubMed/NCBI
4	Gobé G, Browning J, Howard T, Hogg N, Winterford C and Cross R: Apoptosis occurs in endothelial cells during hypertension-induced microvascular rarefaction. J Struct Biol. 118:63–72. 1997. View Article : Google Scholar : PubMed/NCBI
5	White K, Dempsie Y, Caruso P, Wallace E, McDonald RA, Stevens H, Hatley ME, Rooij EV, Morrell NW, MacLean MR and Baker AH: Endothelial apoptosis in pulmonary hypertension is controlled by a microRNA/programmed cell death 4/caspase-3 axis. Hypertension. 64:185–194. 2014. View Article : Google Scholar : PubMed/NCBI
6	Wang ZV, Rothermel BA and Hill JA: Autophagy in hypertensive heart disease. J Biol Chem. 285:8509–8514. 2010. View Article : Google Scholar : PubMed/NCBI
7	Sciarretta S, Maejima Y, Zablocki D and Sadoshima J: The role of autophagy in the heart. Annu Rev Physiol. 80:1–26. 2018. View Article : Google Scholar
8	Nemchenko A, Chiong M, Turer A, Lavandero S and Hill JA: Autophagy as a therapeutic target in cardiovascular disease. J Mol Cell Cardiol. 51:584–593. 2011. View Article : Google Scholar : PubMed/NCBI
9	Wang F, Jia J and Rodrigues B: Autophagy, metabolic disease, and pathogenesis of heart dysfunction. Can J Cardiol. 33:850–859. 2017. View Article : Google Scholar : PubMed/NCBI
10	Savoia C, Battistoni A, Calvez V, Cesario V, Montefusco G and Filippini A: Microvascular alterations in hypertension and vascular aging. Curr Hypertens Rev. 13:16–23. 2017. View Article : Google Scholar : PubMed/NCBI
11	Wu B, Lin J, Luo J, Han D, Fan M, Guo T, Tao L, Yuan M and Yi F: Dihydromyricetin protects against diabetic cardiomyopathy in streptozotocin-induced diabetic mice. Biomed Res Int. 2017:37643702017.PubMed/NCBI
12	Zhang M, Zhang L, Hu J, Lin J, Wang T, Duan Y, Man W, Feng J, Sun L, Jia H, et al: MST1 coordinately regulates autophagy and apoptosis in diabetic cardiomyopathy in mice. Diabetologia. 59:2435–2447. 2016. View Article : Google Scholar : PubMed/NCBI
13	Maejima Y, Kyoi S, Zhai P, Liu T, Li H, Ivessa A, Sciarretta S, Re DPD, Zablocki DK, Hsu CP, et al: Mst1 inhibits autophagy by promoting the interaction between Beclin1 and Bcl-2. Nat Med. 19:1478–1488. 2013. View Article : Google Scholar : PubMed/NCBI
14	Yamamoto S, Yang G, Zablocki D, Liu J, Hong C, Kim SJ, Soler S, Odashima M, Thaisz J, Yehia G, et al: Activation of Mst1 causes dilated cardiomyopathy by stimulating apoptosis without compensatory ventricular myocyte hypertrophy. J Clin Invest. 111:1463–1474. 2003. View Article : Google Scholar : PubMed/NCBI
15	Foulquier S, Namsolleck P, Van Hagen BT, Milanova I, Post MJ, Blankesteijn WM, Rutten BP, Prickaerts J, Van Oostenbrugge RJ and Unger T: Hypertension-induced cognitive impairment: Insights from prolonged angiotensin II infusion in mice. Hypertens Res. 41:817–827. 2018. View Article : Google Scholar : PubMed/NCBI
16	Cheng Z, Zhang M, Hu J, Lin J, Feng X, Wang S, Wang T, Gao E, Wang H and Sun D: Mst1 knockout enhances cardiomyocyte autophagic flux to alleviate angiotensin II-induced cardiac injury independent of angiotensin II receptors. J Mol Cell Cardiol. 125:117–128. 2018. View Article : Google Scholar : PubMed/NCBI
17	Froese N, Kattih B, Breitbart A, Grund A, Geffers R, Molkentin JD, Kispert A, Wollert KC, Drexler H and Heineke J: GATA6 promotes angiogenic function and survival in endothelial cells by suppression of autocrine transforming growth factor beta/activin receptor-like kinase 5 signaling. J Biol Chem. 286:5680–5690. 2011. View Article : Google Scholar
18	Lu Y, Wang RH, Guo BB and Jia YP: Quercetin inhibits angiotensin II induced apoptosis via mitochondrial pathway in human umbilical vein endothelial cells. Eur Rev Med Pharmacol Sci. 20:1609–1616. 2016.PubMed/NCBI
19	He TC, Zhou S, da Costa LT, Yu J, Kinzler KW and Vogelstein B: A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci USA. 95:2509–2514. 1998. View Article : Google Scholar : PubMed/NCBI
20	Van De Vlekkert D, Machado E and d'Azzo A: Analysis of generalized fibrosis in mouse tissue sections with Masson 's Trichrome staining. Bio Protoc. 10:e36292020. View Article : Google Scholar
21	Van Steenkiste C, Trachet B, Casteleyn C, van Loo D, Hoorebeke LV, Segers P, Geerts A, Vlierberghe HV and Colle I: Vascular corrosion casting: Analyzing wall shear stress in the portal vein and vascular abnormalities in portal hypertensive and cirrhotic rodents. Lab Invest. 90:1558–1572. 2010. View Article : Google Scholar : PubMed/NCBI
22	De Biasi S, Gibellini L and Cossarizza A: Uncompensated polychromatic analysis of mitochondrial membrane potential using JC-1 and multilaser excitation. Curr Protoc Cytom. 72:7.32.1–7.32.11. 2015.
23	Arai R and Waguri S: Improved electron microscopy fixation methods for tracking autophagy-associated membranes in cultured mammalian cells. Methods Mol Biol. 1880:211–221. 2019. View Article : Google Scholar : PubMed/NCBI
24	Tomiyama H, Shiina K, Matsumoto-Nakano C, Ninomiya T, Komatsu S, Kimura K, Chikamori T and Yamashina A: The contribution of inflammation to the development of hypertension mediated by increased arterial stiffness. J Am Heart Assoc. 6:e0057292017. View Article : Google Scholar : PubMed/NCBI
25	Kuyumcu F and Aycan A: Evaluation of oxidative stress levels and antioxidant enzyme activities in burst fractures. Med Sci Monit. 24:225–234. 2018. View Article : Google Scholar : PubMed/NCBI
26	Suematsu M, Suzuki H, Delano FA and Schmid-Schonbein GW: The inflammatory aspect of the microcirculation in hypertension: Oxidative stress, leukocytes/endothelial interaction, apoptosis. Microcirculation. 9:259–276. 2002. View Article : Google Scholar : PubMed/NCBI
27	Yang Y, Wang H, Ma Z, Hu W and Sun D: Understanding the role of mammalian sterile 20-like kinase 1 (MST1) in cardiovascular disorders. J Mol Cell Cardiol. 114:141–149. 2018. View Article : Google Scholar
28	Feng X, Wang S, Yang X, Lin J, Man W, Dong Y, Zhang Y, Zhao Z, Wang H and Sun D: Mst1 knockout alleviates mitochondrial fission and mitigates left ventricular remodeling in the development of diabetic cardiomyopathy. Front Cell Dev Biol. 8:6288422020. View Article : Google Scholar
29	Marino G, Niso-Santano M, Baehrecke EH and Kroemer G: Self-consumption: The interplay of autophagy and apoptosis. Nat Rev Mol Cell Biol. 15:81–94. 2014. View Article : Google Scholar : PubMed/NCBI
30	Wilkinson DS, Jariwala JS, Anderson E, Mitra K, Meisenhelder J, Chang JT, Ideker T, Hunter T, Nizet V, Dillin A and Hansen M: Phosphorylation of LC3 by the Hippo kinases STK3/STK4 is essential for autophagy. Mol Cell. 57:55–68. 2015. View Article : Google Scholar :
31	Nieto-Torres JL, Shanahan SL, Chassefeyre R, Chaiamarit T, Zaretski S, Landeras-Bueno S, Verhelle A, Encalada SE and Hansen M: LC3B phosphorylation regulates FYCO1 binding and directional transport of autophagosomes. Curr Biol. 31:3440–3449 e3447. 2021. View Article : Google Scholar : PubMed/NCBI
32	Nieto-Torres JL, Encalada SE and Hansen M: LC3B phosphorylation: Autophagosome's ticket for a ride toward the cell nucleus. Autophagy. 17:3266–3268. 2021. View Article : Google Scholar : PubMed/NCBI
33	Yu W, Xu M, Zhang T, Zhang Q and Zou C: Mst1 promotes cardiac ischemia-reperfusion injury by inhibiting the ERK-CREB pathway and repressing FUNDC1-mediated mitophagy. J Physiol Sci. 69:113–127. 2019. View Article : Google Scholar
34	Wang T, Zhang L, Hu J, Duan Y, Zhang M, Lin J, Man W, Pan X, Jiang Z, Zhang G, et al: Mst1 participates in the atherosclerosis progression through macrophage autophagy inhibition and macrophage apoptosis enhancement. J Mol Cell Cardiol. 98:108–116. 2016. View Article : Google Scholar : PubMed/NCBI
35	Mehta PK and Griendling KK: Angiotensin II cell signaling: Physiological and pathological effects in the cardiovascular system. Am J Physiol Cell Physiol. 292:C82–C97. 2007. View Article : Google Scholar
36	Li AL, Lv JB and Gao L: MiR-181a mediates Ang II-induced myocardial hypertrophy by mediating autophagy. Eur Rev Med Pharmacol Sci. 21:5462–5470. 2017.PubMed/NCBI
37	Kishore R, Krishnamurthy P, Garikipati VN, Benedict C, Nickoloff E, Khan M, Johnson J, Gumpert AM, Koch WJ and Verma SK: Interleukin-10 inhibits chronic angiotensin II-induced pathological autophagy. J Mol Cell Cardiol. 89:203–213. 2015. View Article : Google Scholar : PubMed/NCBI
38	Zhou L, Ma B and Han X: The role of autophagy in angiotensin II-induced pathological cardiac hypertrophy. J Mol Endocrinol. 57:R143–R152. 2016. View Article : Google Scholar : PubMed/NCBI
39	Qin R, Lin D, Zhang L, Xiao F and Guo L: Mst1 deletion reduces hyperglycemia-mediated vascular dysfunction via attenuating mitochondrial fission and modulating the JNK signaling pathway. J Cell Physiol. 235:294–303. 2020. View Article : Google Scholar
40	Dong Q, Xing W, Su F, Liang X, Tian F, Gao F, Wang S and Zhang H: Tetrahydroxystilbene glycoside improves microvascular endothelial dysfunction and ameliorates obesity-associated hypertension in obese ZDF rats via inhibition of endothelial autophagy. Cell Physiol Biochem. 43:293–307. 2017. View Article : Google Scholar : PubMed/NCBI
41	Wang B, Li BW, Li HW, Li AL, Yuan XC, Wang Q and Xiu RJ: Enhanced matrix metalloproteinases-2 activates aortic endothelial hypermeability, apoptosis and vascular rarefaction in spontaneously hypertensive rat. Clin Hemorheol Microcirc. 57:325–338. 2014. View Article : Google Scholar
42	Zorova LD, Popkov VA, Plotnikov EY, Silachev DN, Pevzner IB, Jankauskas SS, Babenko VA, Zorov SD, Balakireva AV, Juhaszova M, et al: Mitochondrial membrane potential. Anal Biochem. 552:50–59. 2018. View Article : Google Scholar :
43	Yan L, Vatner DE, Kim SJ, Ge H, Masurekar M, Massover WH, Yang G, Matsui Y, Sadoshima J and Vatner SF: Autophagy in chronically ischemic myocardium. Proc Natl Acad Sci USA. 102:13807–13812. 2005. View Article : Google Scholar : PubMed/NCBI
44	Nah J, Fernandez AF, Kitsis RN, Levine B and Sadoshima J: Does autophagy mediate cardiac myocyte death during stress? Circ Res. 119:893–895. 2016. View Article : Google Scholar : PubMed/NCBI