Protective efficacy of Shenge San (参蛤散) on mitochondria in H9c2 cardiomyocytes

doi:10.19852/j.cnki.jtcm.2022.06.005

Abstract

Abstract:

OBJECTIVE: To investigate whether peroxisome proliferator-activated receptor γ-coactivator-1α/nuclear respiratory factor 1 (PGC-1α/NRF1) activity can protect mitochondrial function in the setting of cardiac hypertrophy and improve cardiomyocyte energy metabolism.

METHODS: Cardiac hypertrophy was modeled in H9c2 cells treated with isoproterenol (ISO) to assess the effects of Shenge San (参蛤散, SGS) on cell viability and mitochondrial membrane potential. We assessed mitochondrial complex mRNA levels and mitochondrial oxidative phosphorylation factor mRNA and protein levels.

RESULTS: Compared with the 100 μM ISO group, cell size was significantly decreased in the 0.3 mg/mL SGS and 20 μM ZLN005 (PGC-1α activator) groups (P < 0.01). Compared with the SGS (0.3) +ISO group, we observed lower phosphorylated adenosine monophosphate-activated kinase (AMPK) protein levels in the ISO and ZLN005+SGS+ISO groups (P < 0.01). Compared with the compound C group, SGS significantly increased PGC-1α expression in ISO-induced cardiac hypertrophy cells (P < 0.01), and this was inhibited by compound C pretreatment (P < 0.05). Compared with the ISO group, the mitochondrial red-green fluorescence ratio increased in the 0.3 mg/mL SGS group (P < 0.05). mRNA levels of cytochrome c oxidase subunit 1 (CO1) in the ISO and compound C groups were lower than those in control group (P < 0.01), and the mRNA levels of CO1 and ATP8 were significantly lower in the ISO and compound C groups versus control (P < 0.01). Compared with the SGS (0.3) +ISO group, ATP synthetase subunit 8 (ATP8) mRNA was significantly decreased in the ISO group (P < 0.01) and compound C+SGS+ISO group (P < 0.05). Compared with the SGS (0.3) +ISO group, NRF1 mRNA levels were significantly decreased (P < 0.05) in the ISO and compound C+SGS+ISO groups.

CONCLUSIONS: SGS can attenuate ISO-induced cardiomyocyte hypertrophy, restore the decrease in mitochondrial membrane potential, and upregulate PGC-1α/NRF1 levels. Notably, these effects can be blocked by AMPK inhibitor-compound C.

Key words: heart failure, mitochondria, energy metabolism, peroxisome proliferator-activated receptor gamma coactivator 1-alpha, nuclear respiratory factor 1, Shenge San

ZHU Lingyan, WEI Yihong, WANG Youhua, YANG Jianmei, LI Jiawei, CAO Min, ZHOU Duan. Protective efficacy of Shenge San (参蛤散) on mitochondria in H9c2 cardiomyocytes[J]. Journal of Traditional Chinese Medicine, 2022, 42(6): 892-899.

Figures/Tables 4

Figure 1 Comparison of cell size (× 20) A: control group; B: 30 μM ISO for 24 h; C: 100 μM ISO for 24 h; D: 0.3 mg/mL SGS for 1h combined with 100 μM ISO; E: 20 μM ZLN005 for 1 h combined with 100 μM ISO. ISO: isoproterenol; SGS: Shenge San.

Figure 2 Effect of SGS on MMP A1: JC-1 monomer of control group; A2: polymer of control group; A3: merge of control group; B1: JC-1 monomer of ISO (100 μM, 24 h) group; B2: polymer of ISO group; B3: merge of ISO group; C1: JC-1 monomer of ISO + SGS (0.3 mg/mL, 6 h) group; C2: polymer of ISO + SGS group; C3: merge of ISO + SGS group. ISO: isoproterenol; SGS: Shenge San; MMP: mitochondrial membrane potential.

Table 1 Comparison of mRNA expressions of five subunits in mitochondrial complex IV and V between groups

Group	n	CO1	CO2	CO3	ATP6	ATP8
CTRL	10	1.085±0.165	1.067±0.138	1.087±0.154	1.063±0.103	1.113±0.076
Compound C	10	0.703±0.086^a	0.530±0.097^b	0.622±0.110^b	0.778±0.149	0.577±0.092^bc
ISO	10	0.683±0.047^a	0.587±0.064^b	0.567±0.049^b	0.684±0.090^a	0.542±0.070^bd
SGS (0.3) + ISO	10	0.969±0.173	0.783±0.105	0.785±0.136^a	0.943±0.166	0.865±0.126^a
compound C + SGS + ISO	10	0.815±0.039	0.670±0.169^a	0.628±0.023^b	0.681±0.047^a	0.600±0.024^bc

Table 2 Comparison of mRNA expressions of PGC-1α, NRF1, and TFAM between groups

Group	n	PGC-1α	NRF1	TFAM
CTRL	10	1.17±0.20	1.09±0.11	1.11±0.19
Compound C	10	0.62±0.18^a	0.68±0.05^a	0.69±0.14^b
ISO	10	0.66±0.08^b	0.65±0.0827^ac	0.72±0.09^b
SGS (0.3) + ISO	10	0.67±0.07^b	0.93±0.16	0.89±0.10
compound C + SGS + ISO	10	0.58±0.15^a	0.65±0.06^ac	0.68±0.04^b

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