Excessive mitochondrial fission in myocardial ischemia/reperfusion injury: Pathophysiological relevance and potential cardioprotective modulation by plant-derived interventions and electroacupuncture

Table 1 Chemically defined plant-derived compounds associated with mitochondrial fusion-fission regulation in myocardial ischemia/reperfusion injury and related experimental models

Compound	Chemical class	Major plant source	Experimental model	Study groups	Sample size and design	Dosage, administration route, and regimen	Main findings	Evidence strength	Ref.
Ginsenoside Rg1	Triterpenoid saponin	Panax ginseng C.A. Mey (Araliaceae; Ginseng Radix et Rhizoma)	H9c2 H/R (3 hours/3 hours)	Ctrl; H/R; H/R + Rg1	3 independent experiments; R/B: NR	3.125, 6.25, 12.5 μmol/L; MEC NR; pretreated for 24 hours before H/R	Cell viability 66.86% ± 2.655% vs 54.39% ± 0.6338% (H/R; 12.5 μmol/L; mean ± SEM). ↑: MFN2, GDH activity, intracellular glutamate, cell viability, ATP, ΔΨm. ↓: Mitochondrial fragmentation, LDH, CK, ROS, Bax/Bcl-2	C; MFN2 siRNA; Pre (C): Clinical window untested	Dong et al[53]
Ginsenoside Rg1	Triterpenoid saponin	Panax ginseng C.A. Mey (Araliaceae; Ginseng Radix et Rhizoma)	HL-1 H/R (6 hours/2 hours)	Ctrl; H/R; H/R + Rg1; H/R + Rg1 + siNotch1	n = 6 (viability/LDH), 3 (protein assays); R/B: NR	200 μmol/L; MEC NR; added at hypoxia onset and maintained throughout H/R	↑: Notch1, MFN1, ΔΨm, ATP, cell viability. ↓: MFN2, LDH, cytosolic ROS	C; Notch1 siRNA; Isch (C): Clinical window untested	Zhang et al[54]
Ginsenoside Rg5	Triterpenoid saponin	Panax ginseng C.A. Mey (Araliaceae; Ginseng Radix et Rhizoma)	ICR mice, ISO-induced ischemic injury; NRVMs challenged with PA (100 μmol/L, 2 hours), with or without subsequent H/R (4 hours/1 hour)	In vivo: Sham; ISO; ISO + Rg5; TMZ + ISO. In vitro: Ctrl; PA; Rg5 + PA; H/R; PA + H/R; Rg5 + PA + H/R	In vivo: n = 4 (localization), 3-6 (TUNEL). Cells: 3 independent experiments; R/B: NR	In vivo: 50 mg/kg; MED NR; oral; administered 1 hour before ISO on 2 consecutive days. In vitro: 10 μmol/L; MEC NR; pretreated for 0.5 hours before PA	In vivo: ↑: Mitochondrial HK II, and Akt; ↓: Mitochondrial Drp1 recruitment. In vitro: PA model: ↑: P-Akt, mitochondrial HK II, p-Drp1(Ser637), ΔΨm; ↓: Mitochondrial Drp1 recruitment, mPTP opening, ROS, mitochondrial fragmentation. PA + H/R model: ↑: ATP; ↓: Cyt c release, ROS, cell death	C, E; Akt/Drp1 siRNA (cells); Pre: Post-onset untested	Yang et al[55]
Ginsenoside Rc	Triterpenoid saponin	Panax ginseng C.A. Mey (Araliaceae; Ginseng Radix et Rhizoma)	SD rat LAD I/R (2 hours/6 hours); H9c2 H/R (time NR)	In vivo: Sham; I/R; Rc + I/R. In vitro: Ctrl; H/R; Rc + H/R	In vivo: n = 5 (infarct); R: Yes; B: NR	In vivo: 10 mg/kg; MED NR; administered for 14 days before I/R; route and frequency NR. In vitro: 2.5 μmol/L, 5 μmol/L, 10 μmol/L; MEC NR; pretreated for 48 hours before H/R	In vivo: ↑: SIRT1, ETC complex III/IV activities; ↓: Infarct size. In vitro: ↑: SIRT1, ETC complex I-III protein levels, ATP, Bcl-2/Bax; ↓: PGC-1α acetylation, Cyt c	A, C; SIRT1 perturbation (cells); Pre: Post-onset untested	Huang et al[56]
Calenduloside E	Triterpenoid saponin	Aralia elata (Miq) Seem (Araliaceae; NR)	SD rat LAD I/R (30 minutes/48 hours); H9c2 H/R (6 hours/reoxygenation duration NR)	In vivo: Sham, Sham + CE, I/R, I/R + CE. In vitro: Ctrl, H/R, H/R + CE	In vivo: n = 6 (cardiac). Cells: 3 independent experiments; R: Yes (rats); B: NR	In vivo: 15 mg/kg/day; MED NR; gavage for 3 days; last dose 60 minutes before ligation. In vitro: 1 μmol/L, 2 μmol/L, 4 μmol/L, 8 μmol/L; MEC NR; pretreated for 4 hours before hypoxia	In vivo: ↑: MFN2, OPA1, p-AMPK/AMPK, ATP, EF, FS; ↓: Drp1, cTnI, infarct size, Cyt c release, mitochondrial Bax translocation. In vitro: ↑: MFN2, OPA1, p-AMPK/AMPK, ATP, cell viability; ↓: Drp1, Fis1, cleaved caspase-3, cleaved PARP, LDH	A, C; OPA1 siRNA/AMPK blockade (C); Pre: Post-onset untested	Wang et al[60]
Notoginsenoside R1	Triterpenoid saponin	Panax notoginseng (Burkill) F.H. Chen (Araliaceae; Notoginseng Radix et Rhizoma)	H9c2 H/R (6 hours/2 hours)	Ctrl; Ctrl + NGR1; H/R; H/R + NGR1	n = 8 (viability), 6 (LDH); R/B: NR	25 μmol/L; MEC NR; pretreated for 2 hours before H/R and maintained throughout H/R	↑: OPA1, MFN1/2, PGC-1α, NRF1, NRF2, ATP, ETC complexes I-V activity, cell viability. ↓: Drp1, Fis1, mPTP opening, ΔΨm depolarization, ROS, LDH, PINK1, Parkin, BNIP3	C; Assoc.; Pre (C): Clinical window untested	Xu et al[59]
Astragalosidic acid (LS-102)	Triterpenoid saponin derivative	As astragaloside IV derivative; parent compound from Astragalus membranaceus (Fisch) Bunge (Fabaceae; Astragali Radix)	SD rat LAD I/R (30 minutes/90 minutes); H9c2 H/R (12 hours/2 hours)	Sham; I/R; I/R + LS-102; positive control (verapamil). In vitro: Ctrl; H/R; H/R + LS-102	In vivo: n = 6 (infarct), 8-10 (arrhythmia); R: Yes; B: NR	In vivo: 2.5 mg/kg, 5 mg/kg, 10 mg/kg; MED NR; gavage; administered 30 minutes before LAD ligation. In vitro: 0.3125 μmol/L, 0.625 μmol/L, 1.25 μmol/L; MEC NR; pretreated for 24 hours before H/R	VT/VF incidence 10% (1/10) vs 88% (7/8) (I/R; 10 mg/kg). In vivo: ↑: P-PI3K, p-Akt; ↓: Infarct size, CK, LDH In vitro: ↑: P-Drp1(Ser637), p-PI3K, p-Akt, p-GSK-3β(Ser9), ATP, SOD, ΔΨm, Bcl-2/Bax ratio; ↓: P-Drp1(Ser616), CK, LDH, Ca2+ overload, ROS, Cyt c	A, C; PI3K/Akt blockade; Pre: Post-onset untested	Chen et al[58]
Vitexin	Flavonoid glycoside	Crataegus pinnatifida Bunge (Rosaceae; Crataegi Fructus)	SD rat Langendorff I/R (30 minutes/30 minutes); H9c2 H/R (5 hours/2 hours)	Shared (ex vivo/in vitro): Vehicle control; I/R (H/R); VT + I/R (H/R)	Ex vivo: n = 6 (cardiac); cells: n = 4 (protein); R: Yes (both); B: NR	Ex vivo: 10 μmol/L in perfusate; MED NR; perfused for 20 minutes before ischemia and during 30-minute reperfusion. In vitro: 10 μmol/L; MEC NR; pretreated for 2 hours before H/R	Ex vivo: ↑: MFN2, Bcl-2/Bax ratio; ↓: Mitochondrial Drp1, Epac1, Rap1, cytosolic Cyt c, cleaved caspase-3/9, infarct size. In vitro: ↑: MFN2, ΔΨm; ↓: Mitochondrial Drp1, ROS, Epac1, Rap1, NOX4, mitochondrial Bax, cleaved caspase-3/9	B, C; Epac agonist/inhibitor; Pre+Rep (B): Rep-only untested; Pre (C)	Yang et al[67]
Vitexin	Flavonoid glycoside	Crataegus pinnatifida Bunge (Rosaceae; Crataegi Fructus)	SD rat Langendorff I/R (30 minutes/30 minutes); H9c2 H/R (5 hours/1 hour)	Ex vivo: Ctrl; I/R; I/R + VT. In vitro: Ctrl; H/R; H/R + VT	Ex vivo: n = 5-6 (infarct), 6-7 (function); R: Yes (hearts/cells); B: NR	Ex vivo: 1 μmol/L, 3 μmol/L, 10 μmol/L in perfusate; MED NR; perfused for 20 minutes before I/R. In vitro: 1 μmol/L, 3 μmol/L, 10 μmol/L, 30 μmol/L; MEC NR; pretreated for 2 hours before H/R	Ex vivo: ↑: MFN2, COX IV, CF, Bcl-2/Bax ratio; ↓: Mitochondrial Drp1, NOX4, cytosolic Cyt c, cleaved caspase-3/9, infarct size, LDH. In vitro: ↑: MFN2, ATP, ΔΨm; ↓: Mitochondrial Drp1, LDH, cleaved caspase-3/9, ROS	B, C; Assoc; Pre: Post-onset untested	Xue et al[68]
Berberine	Isoquinoline alkaloid	Coptis chinensis Franch (Ranunculaceae; Coptidis Rhizoma)	SD rats ISO-induced injury; H9c2 CoCl2 hypoxia-mimetic injury	In vivo: Vehicle; ISO; ISO + BBR. In vitro: Ctrl; CoCl2; CoCl2 + BBR	In vivo: n = 6 (serum), 3 (protein); cells: n = 3; R: Yes (rats); B: NR	In vivo: 100 mg/kg/day; MED NR; gavage for 14 days before ISO. In vitro: 2.5 μmol/L; MEC NR; pretreated for 24 hours before CoCl2 exposure	In vivo: ↑: MFN2 (mRNA), ATP, Beclin 1, p62, p-AMPK, p-Akt, LKB1, p-Drp1/Drp1; ↓: LDH, CK. In vitro: ↑: Drp1, Beclin 1, p62, p-AMPK, p-Akt, LKB1; ↓: ROS, LDH	E; Assoc; Pre: Post-onset untested	Yang et al[71]
Gastrodin	Phenolic glycoside	Gastrodia elata Blume (Orchidaceae; Gastrodiae Rhizoma)	H9c2 H2O2 oxidative injury (400 μmol/L, 3 hours); additional I/R-like injury (9 hours/3 hours)	H2O2 model: Ctrl; H2O2; GAS + H2O2. I/R-like model: Ctrl; I/R-like; GAS + I/R-like	3 independent experiments; R/B: NR	20 μmol/L, 50 μmol/L, 100 μmol/L; MEC NR; pretreated for 4 hours before injury induction	H2O2 model: ↑: MFN2 and OPA1 mRNA; ↓: Fis1 mRNA, mitochondrial fragmentation, ROS. NRF2-siRNA blunted protection in the H2O2 model; GAS did not significantly alter Drp1 mRNA	C, E; NRF2 siRNA (E); Pre (C,E): Clinical window untested	Cheng et al[62]
Rhein	Anthracenecarboxylic acid	Rheum palmatum L (Polygonaceae; Rhei Radix et Rhizoma)	SD rat LAD-ligation AMI; H9c2 hypoxia 12 hours	Sham; AMI; AMI + rhein; AMI + mdivi-1. In vitro: Ctrl; hypoxia; hypoxia + rhein	In vivo: n = 5 (serum), 3 (TUNEL/protein); R: Yes; B: NR	In vivo: 90 mg/kg; MED NR; gavage for 8 weeks before AMI; frequency NR. In vitro: 1 μg/mL; MEC NR; pretreated for 1 hour before hypoxia	Myocardial TUNEL-positive cells ≈ 15% vs > 30% (AMI; author-reported approximations). Shared (in vivo/in vitro): ↑: OPA1, Beclin 1, PINK1, Parkin, LC3-II/LC3-I; ↓: Drp1, Bax/Bcl-2, cleaved caspase-3. In vivo only: ↓: CK, CK-MB, cTnT, LDH, infarct size	D, E; mdivi-1 comparator; Pre: Post-onset untested	Li et al[73]
Salidroside	Phenylethanoid glycoside	Rhodiola rosea L (Crassulaceae; Rhodiolae roseae Rhizoma et Radix)	SD rat LAD I/R (30 minutes/2 hours); H9c2 H/R (9 hours/6 hours)	In vivo: Sham; I/R; I/R + Sal; I/R + Sal + compound C. In vitro: Ctrl; H/R; H/R + Sal; H/R + Sal + AMPK siRNA	In vivo: n = 6 (infarct/function); R: Yes (rats/cells); B: NR	In vivo: 40 mg/kg; MED NR; administered i.v. 5 minutes before ischemia. In vitro: 10 μmol/L; MEC NR; pretreated for 30 minutes before hypoxia	In vivo: ↑: MFN2, OPA1, p-AMPK/AMPK, ATP, Bcl-2; ↓: P-Drp1(Ser616), Fis1, CHOP, infarct size, LDH, CK, cleaved caspase-3, Bax. In vitro: ↑: P-AMPK/AMPK, Bcl-2; ↓: P-Drp1(Ser616), LDH, CK, CHOP, cleaved caspase-3, Bax	A, C; AMPK blockade/siRNA (C); Pre: Post-onset untested	Tian et al[64]

Selected models, groups and findings are shown; ↑/↓ denote increases/decreases. Minimum effective dose/minimum effective concentration require dose-response support; otherwise not reported. Botanical names and medicinal parts follow authoritative databases and pharmacopoeial/European Medicines Agency monographs. Evidence strength summarizes model directness, mechanistic tests and timing relevance. Model categories: A: In vivo myocardial ischemia/reperfusion (I/R) in an intact animal; B: Isolated-heart I/R without intact systemic context; C: Cellular hypoxia/reoxygenation (R) or oxygen-glucose deprivation/R surrogates; D: Permanent myocardial ischemia/myocardial infarction (MI) without reperfusion; E: Other injury models. A/B directly model cardiac I/R; C models cellular components; D/E provide indirect evidence. Categories are not overall evidence-quality ranks. Assoc: No targeted causal test; named perturbations support the tested pathway, not necessarily fission mediation; comparators do not establish necessity. Pre: Before injury (post-onset initiation untested); Isch: During ischemia; Rep: At/after reperfusion onset. Acute-treatment relevance depends on start time, model, route and exposure; Pre + Rep does not establish Rep-only efficacy. Post-MI: After non-reperfused MI (post-infarction recovery). Cellular timing does not establish clinical windows; timing relevance does not establish clinical efficacy. Sample sizes are endpoint-specific and per group unless independent experiments are specified; cellular n retains the reported replication unit. Only explicitly reported outcome values are shown; graph estimates are excluded. ≈ denotes an approximation stated by the source authors; dispersion is retained as reported. R: Randomization; B: Reported blinding stage; yes: Reported; NR: Not reported; H/R: Hypoxia/reoxygenation; SEM: Standard error of the mean; MFN1/2: Mitofusin 1/2; GDH: Glutamate dehydrogenase; ATP: Adenosine triphosphate; ΔΨm: Mitochondrial membrane potential; LDH: Lactate dehydrogenase; CK: Creatine kinase; ROS: Reactive oxygen species; Bax/Bcl-2: Bcl-2-associated X protein to B-cell lymphoma 2; ISO: Isoproterenol; NRVMs: Neonatal rat ventricular myocytes; PA: Palmitic acid; TMZ: Trimetazidine; HK II: Hexokinase II; Akt: Protein kinase B; Drp1: Dynamin-related protein 1; mPTP: Mitochondrial permeability transition pore; Cyt c: Cytochrome c; LAD: Left anterior descending coronary artery; I/R: Ischemia/reperfusion; SIRT1: Sirtuin 1; ETC: Electron transport chain; PGC-1α: Peroxisome proliferator-activated receptor-γ coactivator 1α; CE: Calenduloside E; OPA1: Optic atrophy 1; AMPK: AMP-activated protein kinase; EF: Ejection fraction; FS: Fractional shortening; cTnI: Cardiac troponin I; PARP: Poly(adenosine diphosphate-ribose) polymerase; NGR1: Notoginsenoside R1; NRF1: Nuclear respiratory factor 1; NRF2: Nuclear factor erythroid 2-related factor 2; PINK1: Phosphatase and tensin homolog-induced kinase 1; BNIP3: B-cell lymphoma 2/adenovirus early region 1B 19-kDa-interacting protein 3; VT/VF: Ventricular tachycardia/ventricular fibrillation; PI3K: Phosphoinositide 3-kinase; GSK-3β: Glycogen synthase kinase 3β; SOD: Superoxide dismutase; Epac1: Exchange protein directly activated by cyclic adenosine monophosphate 1; NOX4: NADPH oxidase 4; BBR: Berberine; LKB1: Liver kinase B1; GAS: Gastrodin; AMI: Acute myocardial infarction; LC3-II: Light chain 3-II; CK-MB: Creatine kinase-MB; cTnT: Cardiac troponin T; Fis1: Mitochondrial fission 1 protein; CHOP: CCAAT/enhancer-binding protein homologous protein.