Ginsenoside Rb1 Ameliorates Autophagy of Hypoxia Cardiomyocytes from Neonatal Rats via AMP-Activated Protein Kinase Pathway
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摘要
Objective: To investigate whether ginsenoside-Rb1(Gs-Rb1) improves the CoCl2-induced autophagy of cardiomyocytes via upregulation of adenosine 5'-monophosphate-activated protein kinase(AMPK) pathway. Methods: Ventricles from 1-to 3-day-old Wistar rats were sequentially digested, separated and incubated in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum for 3 days followed by synchronization. Neonatal rat cardiomyocytes were randomly divided into 7 groups: control group(normal level oxygen), hypoxia group(500 μmol/L CoCl_2), Gs-Rb1 group(200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2), Ara A group(500 μmol/L Ara A + 500 μmol/L CoCl_2), Ara A+ Gs-Rb1 group(500 μmol/L Ara A + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2), AICAR group [1 mmol/L 5-aminoimidazole-4-carboxamide ribonucleotide(AICAR) + 500 μmol/L CoCl_2], and AICAR+Gs-Rb1 group(1 mmol/L AICAR + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2). Cel s were treated for 12 h and cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay and cardiac troponin I(cTnI) levels were detected by enzyme-linked immunosorbent assay(ELISA). AMPK activity was assessed by 2',7'-dichlorofluorescein diacetate(DCFH-DA) ELISA assay. The protein expressions of Atg4 B, Atg5, Atg6, Atg7, microtubule-associated protein 1 A/1 B-light chain 3(LC3), P62, and active-cathepsin B were measured by Western blot. Results: Gs-Rb1 significantly improved the cell viability of hypoxia cardiomyocytes(P<0.01). However, the viability of hypoxia-treated cardiomyocytes was significantly inhibited by Ara A(P<0.01). Gs-Rb1 increased the AMPK activity of hypoxia-treated cardiomyocytes. The AMPK activity of hypoxia-treated cadiomyocytes was inhibited by Ara A(P<0.01) and was not affected by AICAR(P=0.983). Gs-Rb1 up-regulated Atg4B, Atg5, Beclin-1, Atg7, LC3B Ⅱ, the LC3BⅡ/Ⅰ ratio and cathepsin B activity of hypoxia cardiomyocytes(P<0.05), each of these protein levels was significantly enhanced by Ara A(all P<0.01), but was not affected by AICAR(all P>0.05). Gs-Rb1 significantly down-regulated P62 levels of hypoxic cardiomyocytes(P<0.05). The P62 levels of hypoxic cardiomyocytes were inhibited by Ara A(P<0.05) and were not affected by AICAR(P=0.871). Conclusion: Gs-Rb1 may improve the viability of hypoxia cardiomyocytes by ameliorating cell autophagy via the upregulation of AMPK pathway.
Objective: To investigate whether ginsenoside-Rb1(Gs-Rb1) improves the CoCl2-induced autophagy of cardiomyocytes via upregulation of adenosine 5'-monophosphate-activated protein kinase(AMPK) pathway. Methods: Ventricles from 1-to 3-day-old Wistar rats were sequentially digested, separated and incubated in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum for 3 days followed by synchronization. Neonatal rat cardiomyocytes were randomly divided into 7 groups: control group(normal level oxygen), hypoxia group(500 μmol/L CoCl_2), Gs-Rb1 group(200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2), Ara A group(500 μmol/L Ara A + 500 μmol/L CoCl_2), Ara A+ Gs-Rb1 group(500 μmol/L Ara A + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2), AICAR group [1 mmol/L 5-aminoimidazole-4-carboxamide ribonucleotide(AICAR) + 500 μmol/L CoCl_2], and AICAR+Gs-Rb1 group(1 mmol/L AICAR + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2). Cel s were treated for 12 h and cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay and cardiac troponin I(cTnI) levels were detected by enzyme-linked immunosorbent assay(ELISA). AMPK activity was assessed by 2',7'-dichlorofluorescein diacetate(DCFH-DA) ELISA assay. The protein expressions of Atg4 B, Atg5, Atg6, Atg7, microtubule-associated protein 1 A/1 B-light chain 3(LC3), P62, and active-cathepsin B were measured by Western blot. Results: Gs-Rb1 significantly improved the cell viability of hypoxia cardiomyocytes(P<0.01). However, the viability of hypoxia-treated cardiomyocytes was significantly inhibited by Ara A(P<0.01). Gs-Rb1 increased the AMPK activity of hypoxia-treated cardiomyocytes. The AMPK activity of hypoxia-treated cadiomyocytes was inhibited by Ara A(P<0.01) and was not affected by AICAR(P=0.983). Gs-Rb1 up-regulated Atg4B, Atg5, Beclin-1, Atg7, LC3B Ⅱ, the LC3BⅡ/Ⅰ ratio and cathepsin B activity of hypoxia cardiomyocytes(P<0.05), each of these protein levels was significantly enhanced by Ara A(all P<0.01), but was not affected by AICAR(all P>0.05). Gs-Rb1 significantly down-regulated P62 levels of hypoxic cardiomyocytes(P<0.05). The P62 levels of hypoxic cardiomyocytes were inhibited by Ara A(P<0.05) and were not affected by AICAR(P=0.871). Conclusion: Gs-Rb1 may improve the viability of hypoxia cardiomyocytes by ameliorating cell autophagy via the upregulation of AMPK pathway.
Objective: To investigate whether ginsenoside-Rb1(Gs-Rb1) improves the CoCl2-induced autophagy of cardiomyocytes via upregulation of adenosine 5'-monophosphate-activated protein kinase(AMPK) pathway. Methods: Ventricles from 1-to 3-day-old Wistar rats were sequentially digested, separated and incubated in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum for 3 days followed by synchronization. Neonatal rat cardiomyocytes were randomly divided into 7 groups: control group(normal level oxygen), hypoxia group(500 μmol/L CoCl_2), Gs-Rb1 group(200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2), Ara A group(500 μmol/L Ara A + 500 μmol/L CoCl_2), Ara A+ Gs-Rb1 group(500 μmol/L Ara A + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2), AICAR group [1 mmol/L 5-aminoimidazole-4-carboxamide ribonucleotide(AICAR) + 500 μmol/L CoCl_2], and AICAR+Gs-Rb1 group(1 mmol/L AICAR + 200 μmol/L Gs-Rb1 + 500 μmol/L CoCl_2). Cel s were treated for 12 h and cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide(MTT) assay and cardiac troponin I(cTnI) levels were detected by enzyme-linked immunosorbent assay(ELISA). AMPK activity was assessed by 2',7'-dichlorofluorescein diacetate(DCFH-DA) ELISA assay. The protein expressions of Atg4 B, Atg5, Atg6, Atg7, microtubule-associated protein 1 A/1 B-light chain 3(LC3), P62, and active-cathepsin B were measured by Western blot. Results: Gs-Rb1 significantly improved the cell viability of hypoxia cardiomyocytes(P<0.01). However, the viability of hypoxia-treated cardiomyocytes was significantly inhibited by Ara A(P<0.01). Gs-Rb1 increased the AMPK activity of hypoxia-treated cardiomyocytes. The AMPK activity of hypoxia-treated cadiomyocytes was inhibited by Ara A(P<0.01) and was not affected by AICAR(P=0.983). Gs-Rb1 up-regulated Atg4B, Atg5, Beclin-1, Atg7, LC3B Ⅱ, the LC3BⅡ/Ⅰ ratio and cathepsin B activity of hypoxia cardiomyocytes(P<0.05), each of these protein levels was significantly enhanced by Ara A(all P<0.01), but was not affected by AICAR(all P>0.05). Gs-Rb1 significantly down-regulated P62 levels of hypoxic cardiomyocytes(P<0.05). The P62 levels of hypoxic cardiomyocytes were inhibited by Ara A(P<0.05) and were not affected by AICAR(P=0.871). Conclusion: Gs-Rb1 may improve the viability of hypoxia cardiomyocytes by ameliorating cell autophagy via the upregulation of AMPK pathway.
引文
1.Shintani T,Klionsky DJ.Autophagy in health and disease:a double-edged sword.Science 2004;306:990-995.
2.Yan L,Vatner DE,Kim SJ,Ge H,Masurekar M,Massover WH,et al.Autophagy in chronically ischemic myocardium.Proc Natl Acad Sci U S A 2005;12:13807-13812.
3.Hamacher-Brady A,Brady NR,Gottlieb RA.Enhancing macroautophagy protects against ischemia/reperfusion injury in cardiac myocytes.J Biol Chem 2006;28:29776-29787.
4.Dosenko VE,Nagibin VS,Tumanovska LV,Moibenko AA.Protective effect of autophagy in anoxia-reoxygenation of isolated cardiomyocyte.Autophagy 2006;2:305-306.
5.Matsui Y,Takagi H,Qu X,Abdellatif M,Sakoda H,Asano T,et al.Distinct roles of autophagy in the heart during ischemia and reperfusion:roles of AMP-activated protein kinase and Beclin 1 in mediating autophagy.Circ Res 2007;100:914-922.
6.Gui L,Liu B,Lu G.Hypoxia induces autophagy in cardiomyocytes via a hypoxia-inducible factor 1-dependent mechanism.Exp Ther Med 2016;11:2233-2239.
7.Jiang QS,Huang XN,Yang GZ,Jiang XY,Zhou QX.Inhibitory effect of ginsenoside Rb1 on calcineurin signal pathway in cardiomyocyte hypertrophy induced by prostaglandin F2alpha.Acta Pharmacol Sin(Chin)2007;28:1149-1154.
8.Kong HL,Wang JP,Li ZQ,Zhao SM,Dong J,Zhang WW.Anti-hypoxic effect of ginsenoside Rb1 on neonatal rat cardiomyocytes is mediated through the specific activation of glucose transporter-4 ex vivo.Acta Pharmacol Sin(Chin)2009;30:396-403.
9.Kong HL,Li ZQ,Zhao YJ,Zhao SM,Zhu L,Li T,et al.Ginsenoside Rb1 protects cardiomyocytes against CoCl2-induced apoptosis in neonatal rats by inhibiting mitochondria permeability transition pore opening.Acta Pharmacol Sin(Chin)2010;31:687-695.
10.Kong HL,Li ZQ,Zhao SM,Yuan L,Miao ZL,Liu Y,et al.Apelin-APJ effects of ginsenoside-Rb1 depending on hypoxia-induced factor 1αin hypoxia neonatal cardiomyocytes.Chin J Integr Med 2015;21:139-146.
11.Boya P,Gonzalez-Polo RA,Casares N,Perfettini JL,Dessen P,Larochette N,et al.Inhibition of macroautophagy triggers apoptosis.Mol Cell Biol 2005;25:1025-1040.
12.Kim I,Rodriguez-Enriquez S,Lemasters JJ.Selective degradation of mitochondria by mitophagy.Arch Biochem Biophys 2007;462:245-253.
13.Levine B,Yuan J.Autophagy in cell death:an innocent convict?J Clin Invest 2005;115:2679-2688.
14.Thorburn A.Apoptosis and autophagy:regulatory connections between two supposedly different processes.Apoptosis 2008;13:1-9.
15.Maiuri MC,Zalckvar E,Kimchi A,Kroemer G.Selfeating and self-killing:crosstalk between autophagy and apoptosis.Nat Rev Mol Cell Biol 2007;8:741-752.
16.Espert L,Denizot M,Grimaldi M,Robert-Hebmann V,Gay B,Varbanov M,et al.Autophagy is involved in T cell death after binding of HIV-1 envelope proteins to CXCR4.J Clin Invest 2006;116:2161-2172.
17.Moretti L,Cha YI,Niermann KJ,Lu B.Switch between apoptosis and autophagy:radiation-induced endoplasmic reticulum stress?Cell Cycle 2007;6:793-798.
18.Boyce M,Yuan J.Cellular response to endoplasmic reticulum stress:a matter of life or death.Cell Death Differ2006;13:363-373.
19.Kimura S,Noda T,Yoshimori T.Dissection of the autophagosome maturation process by a novel reporter protein,tandem fluorescent-tagged LC3.Autophagy2007;3:452-460.
20.Xie Z,Nair U,Klionsky DJ.Atg8 controls phagophore expansion during autophagosome formation.Mol Biol Cell2008;19:3290-3298.
21.Mizushima N,Yamamoto A,Hatano M,Kobayashi Y,Kabeya Y,Suzuki K,et al.Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells.J Cell Biol 2001;152:657-668.
22.Kihara A,Kabeya Y,Ohsumi Y,Yoshimori T.Beclinphosphatidylinositol 3-kinase complex functions at the trans-Golgi network.EMBO Rep 2001;2:330-335.
23.Matsui Y,Takagi H,Qu X,Abdellatif M,Sakoda H,Asano T,et al.Distinct roles of autophagy in the heart during ischemia and reperfusion:roles of AMP-activated protein kinase and Beclin 1 in mediating autophagy.Circ Res 2007;100:914-922.
24.Valentim L,Laurence KM,Townsend PA,Carroll CJ,Soond S,Scarabelli TM,et al.Urocortin inhibits Beclin1-mediated autophagic cell death in cardiac myocytes exposed to ischaemia/reperfusion injury.J Mol Cell Cardiol2006;40:846-852.
25.Ma X,Liu H,Foyil SR,Godar RJ,Weinheimer CJ,Hill JA,et al.Impaired autophagosome clearance contributes to cardiomyocyte death in ischemia/reperfusion injury.Circulation 2012;125:3170-3181.
26.Zeng M,Wei X,Wu Z,Li W,Li B,Zhen Y,et al.NF-κB-mediated induction of autophagy in cardiac ischemia/reperfusion injury.Biochem Biophys Res Commun2013;436:180-185.
27.Feng Y,Yao Z,Klionsky DJ.How to control self-digestion:transcriptional,post-transcriptional,and post-translational regulation of autophagy.Trends Cell Biol 2015;25:354-363.
28.Nakatogawa H,Suzuki K,Kamada Y,Ohsumi Y.Dynamics and diversity in autophagy mechanisms:lessons from yeast.Nat Rev Mol Cell Biol 2009;10:458-467.
29.Shintani T,Huang WP,Stromhaug PE,Klionsky DJ.Mechanism of cargo selection in the cytoplasm to vacuole targeting pathway.Dev Cell 2002;3:825-837.
30.Xie Z,Nair U,Klionsky DJ.Atg8 controls phagophore expansion during autophagosome formation.Mol Biol Cell2008;19:3290-3298.
31.Kaufmann A,Beier V,Franquelim HG,Wollert T.Molecular mechanism of autophagic membrane-scaffold assembly and disassembly.Cell 2014;156:469-481.
32.Zhang J,He Z,Xiao W,Na Q,Wu T,Su K,et al.Overexpression of BAG3 attenuates hypoxia-induced cardiomyocyte apoptosis by inducing autophagy.Cell Physiol Biochem 2016;39:491-500.
33.Gao YH,Qian JY,Chen ZW,Fu MQ,Xu JF,Xia Y,et al.Suppression of Bim by microRNA-19a may protect cardiomyocytes against hypoxia-induced cell death via autophagy activation.Toxicol Lett 2016;257:72-83.
34.Hsieh DJ,Kuo WW,Lai YP,Shibu MA,Shen CY,Pai P,et al.17β-Estradiol and/or estrogen receptorβattenuate the autophagic and apoptotic effects induced by prolonged hypoxia through HIF-1α-mediated BNIP3 and IGFBP-3signaling blockage.Cell Physiol Biochem 2015;36:274-284.
35.Komatsu M,Kageyama S,Ichimura Y.P62/SQSTM1/A170:physiology and pathology.Pharmacol Res 2012;66:457-462.
36.Layfield R,Cavey JR,Najat D,Long J,Sheppard PW,Ralston SH,et al.p62 mutations,ubiquitin recognition and Paget's disease of bone.Bio Soc Trans 2006;34:735-737.
37.Lee JH,Yu WH,Kumar A,Lee S,Mohan PS,Peterhoff CM,et al.Lysosomal proteolysis and autophagy require presenilin 1 and are disrupted by Alzheimer-related PS1mutations.Cell 2010;141:1146-1158.
38.Moscat J,Diaz-Meco MT,Wooten MW.Signal integration and diversification through the p62 scaffold protein.Trends Biol Sci 2007;32:95-100.
39.Pankiv S,Clausen TH,Lamark T,Brech A,Bruun JA,Outzen H,et al.p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy.J Biol Chem 2007;282:24131-24145.
40.Mizushima N,Hara T.Intracellular quality control by autophagy:how does autophagy prevent neurodegeneration?Autophagy 2006;2:302-304.
41.Bj?rk?y G,Lamark T,Brech A,Outzen H,Perander M,Overvatn A,et al.p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death.J Cell Biol 2005;171:603-614.
42.Wang Q,Yang L,Hua Y,Nair S,Xu X,Ren J.AMP-activated protein kinase deficiency rescues paraquat-induced cardiac contractile dysfunction through an autophagy-dependent mechanism.Toxicol Sci 2014;142:6-20.
43.Inoki K,Zhu T,Guan KL.TSC2 mediates cellular energy response to control cell growth and survival.Cell2003;115:577-590.
44.Tripathi DN,Chowdhury R,Trudel LJ,Tee AR,Slack RS,Walker CL,et al.Reactive nitrogen species regulate autophagy through ATM-AMPK-TSC 2-mediated suppression of mTORC1.Proc Natl Acad Sci U S A2013;110:E2950-E2957.
2.Yan L,Vatner DE,Kim SJ,Ge H,Masurekar M,Massover WH,et al.Autophagy in chronically ischemic myocardium.Proc Natl Acad Sci U S A 2005;12:13807-13812.
3.Hamacher-Brady A,Brady NR,Gottlieb RA.Enhancing macroautophagy protects against ischemia/reperfusion injury in cardiac myocytes.J Biol Chem 2006;28:29776-29787.
4.Dosenko VE,Nagibin VS,Tumanovska LV,Moibenko AA.Protective effect of autophagy in anoxia-reoxygenation of isolated cardiomyocyte.Autophagy 2006;2:305-306.
5.Matsui Y,Takagi H,Qu X,Abdellatif M,Sakoda H,Asano T,et al.Distinct roles of autophagy in the heart during ischemia and reperfusion:roles of AMP-activated protein kinase and Beclin 1 in mediating autophagy.Circ Res 2007;100:914-922.
6.Gui L,Liu B,Lu G.Hypoxia induces autophagy in cardiomyocytes via a hypoxia-inducible factor 1-dependent mechanism.Exp Ther Med 2016;11:2233-2239.
7.Jiang QS,Huang XN,Yang GZ,Jiang XY,Zhou QX.Inhibitory effect of ginsenoside Rb1 on calcineurin signal pathway in cardiomyocyte hypertrophy induced by prostaglandin F2alpha.Acta Pharmacol Sin(Chin)2007;28:1149-1154.
8.Kong HL,Wang JP,Li ZQ,Zhao SM,Dong J,Zhang WW.Anti-hypoxic effect of ginsenoside Rb1 on neonatal rat cardiomyocytes is mediated through the specific activation of glucose transporter-4 ex vivo.Acta Pharmacol Sin(Chin)2009;30:396-403.
9.Kong HL,Li ZQ,Zhao YJ,Zhao SM,Zhu L,Li T,et al.Ginsenoside Rb1 protects cardiomyocytes against CoCl2-induced apoptosis in neonatal rats by inhibiting mitochondria permeability transition pore opening.Acta Pharmacol Sin(Chin)2010;31:687-695.
10.Kong HL,Li ZQ,Zhao SM,Yuan L,Miao ZL,Liu Y,et al.Apelin-APJ effects of ginsenoside-Rb1 depending on hypoxia-induced factor 1αin hypoxia neonatal cardiomyocytes.Chin J Integr Med 2015;21:139-146.
11.Boya P,Gonzalez-Polo RA,Casares N,Perfettini JL,Dessen P,Larochette N,et al.Inhibition of macroautophagy triggers apoptosis.Mol Cell Biol 2005;25:1025-1040.
12.Kim I,Rodriguez-Enriquez S,Lemasters JJ.Selective degradation of mitochondria by mitophagy.Arch Biochem Biophys 2007;462:245-253.
13.Levine B,Yuan J.Autophagy in cell death:an innocent convict?J Clin Invest 2005;115:2679-2688.
14.Thorburn A.Apoptosis and autophagy:regulatory connections between two supposedly different processes.Apoptosis 2008;13:1-9.
15.Maiuri MC,Zalckvar E,Kimchi A,Kroemer G.Selfeating and self-killing:crosstalk between autophagy and apoptosis.Nat Rev Mol Cell Biol 2007;8:741-752.
16.Espert L,Denizot M,Grimaldi M,Robert-Hebmann V,Gay B,Varbanov M,et al.Autophagy is involved in T cell death after binding of HIV-1 envelope proteins to CXCR4.J Clin Invest 2006;116:2161-2172.
17.Moretti L,Cha YI,Niermann KJ,Lu B.Switch between apoptosis and autophagy:radiation-induced endoplasmic reticulum stress?Cell Cycle 2007;6:793-798.
18.Boyce M,Yuan J.Cellular response to endoplasmic reticulum stress:a matter of life or death.Cell Death Differ2006;13:363-373.
19.Kimura S,Noda T,Yoshimori T.Dissection of the autophagosome maturation process by a novel reporter protein,tandem fluorescent-tagged LC3.Autophagy2007;3:452-460.
20.Xie Z,Nair U,Klionsky DJ.Atg8 controls phagophore expansion during autophagosome formation.Mol Biol Cell2008;19:3290-3298.
21.Mizushima N,Yamamoto A,Hatano M,Kobayashi Y,Kabeya Y,Suzuki K,et al.Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells.J Cell Biol 2001;152:657-668.
22.Kihara A,Kabeya Y,Ohsumi Y,Yoshimori T.Beclinphosphatidylinositol 3-kinase complex functions at the trans-Golgi network.EMBO Rep 2001;2:330-335.
23.Matsui Y,Takagi H,Qu X,Abdellatif M,Sakoda H,Asano T,et al.Distinct roles of autophagy in the heart during ischemia and reperfusion:roles of AMP-activated protein kinase and Beclin 1 in mediating autophagy.Circ Res 2007;100:914-922.
24.Valentim L,Laurence KM,Townsend PA,Carroll CJ,Soond S,Scarabelli TM,et al.Urocortin inhibits Beclin1-mediated autophagic cell death in cardiac myocytes exposed to ischaemia/reperfusion injury.J Mol Cell Cardiol2006;40:846-852.
25.Ma X,Liu H,Foyil SR,Godar RJ,Weinheimer CJ,Hill JA,et al.Impaired autophagosome clearance contributes to cardiomyocyte death in ischemia/reperfusion injury.Circulation 2012;125:3170-3181.
26.Zeng M,Wei X,Wu Z,Li W,Li B,Zhen Y,et al.NF-κB-mediated induction of autophagy in cardiac ischemia/reperfusion injury.Biochem Biophys Res Commun2013;436:180-185.
27.Feng Y,Yao Z,Klionsky DJ.How to control self-digestion:transcriptional,post-transcriptional,and post-translational regulation of autophagy.Trends Cell Biol 2015;25:354-363.
28.Nakatogawa H,Suzuki K,Kamada Y,Ohsumi Y.Dynamics and diversity in autophagy mechanisms:lessons from yeast.Nat Rev Mol Cell Biol 2009;10:458-467.
29.Shintani T,Huang WP,Stromhaug PE,Klionsky DJ.Mechanism of cargo selection in the cytoplasm to vacuole targeting pathway.Dev Cell 2002;3:825-837.
30.Xie Z,Nair U,Klionsky DJ.Atg8 controls phagophore expansion during autophagosome formation.Mol Biol Cell2008;19:3290-3298.
31.Kaufmann A,Beier V,Franquelim HG,Wollert T.Molecular mechanism of autophagic membrane-scaffold assembly and disassembly.Cell 2014;156:469-481.
32.Zhang J,He Z,Xiao W,Na Q,Wu T,Su K,et al.Overexpression of BAG3 attenuates hypoxia-induced cardiomyocyte apoptosis by inducing autophagy.Cell Physiol Biochem 2016;39:491-500.
33.Gao YH,Qian JY,Chen ZW,Fu MQ,Xu JF,Xia Y,et al.Suppression of Bim by microRNA-19a may protect cardiomyocytes against hypoxia-induced cell death via autophagy activation.Toxicol Lett 2016;257:72-83.
34.Hsieh DJ,Kuo WW,Lai YP,Shibu MA,Shen CY,Pai P,et al.17β-Estradiol and/or estrogen receptorβattenuate the autophagic and apoptotic effects induced by prolonged hypoxia through HIF-1α-mediated BNIP3 and IGFBP-3signaling blockage.Cell Physiol Biochem 2015;36:274-284.
35.Komatsu M,Kageyama S,Ichimura Y.P62/SQSTM1/A170:physiology and pathology.Pharmacol Res 2012;66:457-462.
36.Layfield R,Cavey JR,Najat D,Long J,Sheppard PW,Ralston SH,et al.p62 mutations,ubiquitin recognition and Paget's disease of bone.Bio Soc Trans 2006;34:735-737.
37.Lee JH,Yu WH,Kumar A,Lee S,Mohan PS,Peterhoff CM,et al.Lysosomal proteolysis and autophagy require presenilin 1 and are disrupted by Alzheimer-related PS1mutations.Cell 2010;141:1146-1158.
38.Moscat J,Diaz-Meco MT,Wooten MW.Signal integration and diversification through the p62 scaffold protein.Trends Biol Sci 2007;32:95-100.
39.Pankiv S,Clausen TH,Lamark T,Brech A,Bruun JA,Outzen H,et al.p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy.J Biol Chem 2007;282:24131-24145.
40.Mizushima N,Hara T.Intracellular quality control by autophagy:how does autophagy prevent neurodegeneration?Autophagy 2006;2:302-304.
41.Bj?rk?y G,Lamark T,Brech A,Outzen H,Perander M,Overvatn A,et al.p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death.J Cell Biol 2005;171:603-614.
42.Wang Q,Yang L,Hua Y,Nair S,Xu X,Ren J.AMP-activated protein kinase deficiency rescues paraquat-induced cardiac contractile dysfunction through an autophagy-dependent mechanism.Toxicol Sci 2014;142:6-20.
43.Inoki K,Zhu T,Guan KL.TSC2 mediates cellular energy response to control cell growth and survival.Cell2003;115:577-590.
44.Tripathi DN,Chowdhury R,Trudel LJ,Tee AR,Slack RS,Walker CL,et al.Reactive nitrogen species regulate autophagy through ATM-AMPK-TSC 2-mediated suppression of mTORC1.Proc Natl Acad Sci U S A2013;110:E2950-E2957.