The relationship between the weakened myocardial protective effect of sevoflurane pretreatment by high glucose factors and the HIF-1&#x03B1;/LDHA signaling pathway

doi:10.3877/cma.j.issn.2095-1221.2026.04.004

Abstract

Abstract:

Objective

To investigate whether the mechanism by which high glucose weaken the protective effect of sevoflurane pretreatment on rat cardiomyocytes is related to the HIF-1α/LDHA signaling pathway.

Methods

H9c2 cardiomyocytes were randomly divided into 8 groups using a random number table, and named control group (N+Con group), hypoxia-reoxygenation group (N+H/R group), sevoflurane preconditioning group (N+SPC group), high glucose control group (H+Con group), high glucose-hypoxia-reoxygenation group (H+H/R group), high glucose sevoflurane preconditioning group (H+SPC group), high glucose sevoflurane preconditioning + HIF-1α agonist group (H+SPC+D group), and high glucose sevoflurane preconditioning + HIF-1α agonist + LDHA inhibitor group (H+SPC+D+A group). Cell viability of cardiomyocytes was assessed using the CCK-8 assay. Lactate dehydrogenase (LDH) levels in the culture medium were measured via the microplate method. Lactate levels were determined using the WST-8 assay. ATP levels were detected by chemiluminescence assay. The levels of key glycolytic enzymes were quantified using ELISA. Apoptosis rate was evaluated by flow cytometry, and mitochondrial membrane potential was measured using a fluorescent probe. The expression levels of HIF-1α and LDHA were detected by western blotting. Comparisons among multiple groups were performed using one-way ANOVA, and pairwise comparisons between groups were conducted using Tukey's multiple comparison test.

Results

Compared with the N+SPC group, the H+SPC group showed decreases in cell viability [(70.12 ± 3.81)%vs (86.66 ± 5.48)%], lactate level [(5.36 ± 0.18) vs (6.94 ± 0.45) mmol/L], ATP level [(88.90 ± 17.33) vs (172.88 ± 26.83) μmol/L], mitochondrial membrane potential (0.55 ± 0.11 vs 0.78 ± 0.08), the activities of HK, PFK1, and PK [(72.43 ± 8.77) vs (134.23 ± 8.84) U/mL, (104.62 ± 10.65) vs (213.30 ± 12.36) U/mL, (92.87 ± 5.14) vs (251.67 ± 14.45) U/mL], the expression of HIF-1α (0.54 ± 0.01 vs 0.94 ± 0.04) and LDHA (0.69 ± 0.02 vs 1.07 ± 0.04) were decreased in the H+SPC group, while the LDH level in culture medium [(315.12 ± 25.22) vs (122.87 ± 8.03) U/L ] and apoptosis rate were increased [(21.42 ± 2.11)% vs (10.92 ± 0.69)%]. All differences were statistically significant (all P < 0.05). Compared with the H+SPC group, the cell viability [(83.72 ± 4.55)%vs (70.12 ± 3.81)%], lactate level [(6.40 ± 0.42) vs (5.36 ± 0.18) mmol/L], ATP level [(165.62 ± 24.28) vs (88.90 ± 17.33) μmol/L], mitochondrial membrane potential (0.82 ± 0.05 vs 0.55 ± 0.11), the activities of HK, PFK1, and PK [(122.34 ± 8.74 vs 72.43 ± 8.77) U/mL, (192.45 ± 20.98) vs (104.62 ± 10.65) U/mL, (208.76 ± 32.65) vs (92.87 ± 5.14) U/mL], the expression of HIF-1α (0.80 ± 0.03 vs 0.54 ± 0.01) and LDHA (0.87 ± 0.04 vs 0.69 ± 0.02) were increased in the H+SPC+D group, whereas culture medium LDH level [(154.33 ± 26.85) vs (315.12 ± 25.22) U/L ] and apoptosis rate [(15.43 ± 1.07)% vs (21.42 ± 2.11)%] were decreased. All differences were statistically significant (all P < 0.05). Compared with the H+SPC+D group, the cell viability [(68.43 ± 3.20)% vs (83.72 ± 4.55)%], lactate level [(5.03 ± 0.12) vs (6.40 ± 0.42) mmol/L], ATP level [(79.54 ± 19.22) vs (165.62 ± 24.28) μmol/L], mitochondrial membrane potential (0.57 ± 0.05 vs 0.82 ± 0.05), the activities of HK, PFK1, and PK [(66.83 ± 6.91) vs (122.34 ± 8.74) U/mL, (77.83 ± 5.94) vs (192.45 ± 20.98) U/mL, (132.80 ± 3.53) vs (208.76 ± 32.65) U/mL], the expression of HIF-1α (0.35 ± 0.02 vs 0.80 ± 0.03) and LDHA (0.45 ± 0.02 vs 0.87 ± 0.04) were decreased in the H+SPC+D+A group, while culture medium LDH level [(264.96 ± 13.46) vs (154.33 ± 26.85)U/L] and apoptosis rate [(19.83 ± 0.23)%vs (15.43 ± 1.07)%] were increased. All differences were statistically significant (all P < 0.05).

Conclusion

The mechanism by which high glucose attenuates the protective effect of sevoflurane preconditioning on rat cardiomyocytes may be related to the inhibition of HIF-1α/LDHA signaling pathway activity.

Key words: High glucose, H9c2 cells, Hypoxia/reoxygenation, Hypoxia inducible factor 1α, Lactate dehydrogenase A

Tianliang Hou, Haixia Wang, Haiping Ma, Jianrong Ye. The relationship between the weakened myocardial protective effect of sevoflurane pretreatment by high glucose factors and the HIF-1α/LDHA signaling pathway[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2026, 16(04): 218-226.

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URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2026.04.004

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Figures/Tables 6

References 21

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分组	细胞活力（%）	培养液LDH水平（U/L）	乳酸水平（mmol/L）	ATP水平（μmol/L）	凋亡率（%）	线粒体膜电位
N+Con组	100.03 ± 4.67	103.82 ± 14.08	3.75 ± 0.43	266.74 ± 26.08	5.11 ± 1.01	1.21 ± 0.02
N+H/R组	71.23 ± 4.23^a	275.96 ± 12.32^a	5.93 ± 0.19^a	86.61 ± 18.72^a	21.69 ± 0.37^a	0.54 ± 0.04^a
N+SPC组	86.66 ± 5.48^b	122.87 ± 8.03^b	6.94 ± 0.45^ab	172.88 ± 26.83^ab	10.92 ± 0.69^ab	0.78 ± 0.08^ab
H+Con组	91.15 ± 4.32^b	252.15 ± 24.19^abc	3.58 ± 0.33^bc	247.63 ± 21.45^abc	8.33 ± 0.07^abc	0.94 ± 0.02^ab
H+H/R组	65.74 ± 7.35^acd	367.52 ± 43.63^abcd	5.33 ± 0.22^abcd	38.59 ± 22.99^abcd	25.12 ± 0.46^abcd	0.46 ± 0.02^acd
H+SPC组	70.12 ± 3.81^acd	315.12 ± 25.22^abcde	5.36 ± 0.18^abcd	88.90 ± 17.33^acde	21.42 ± 2.11^acde	0.55 ± 0.11^acd
H+SPC+D组	83.72 ± 4.55^abef	154.33 ± 26.85^abdef	6.40 ± 0.42^abcdef	165.62 ± 24.28^abdef	15.43 ± 1.07^abcdef	0.82 ± 0.05^abef
H+SPC+D+A组	68.43 ± 3.20^acdg	264.96 ± 13.46^aceg	5.03 ± 0.12^abcdg	79.54 ± 19.22^acdeg	19.83 ± 0.23^acdeg	0.57 ± 0.05^acdg
F值	21.211	32.012	125.517	173.711	221.412	40.473
P值	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

分组	细胞活力（%）	培养液LDH水平（U/L）	乳酸水平（mmol/L）	ATP水平（μmol/L）	凋亡率（%）	线粒体膜电位
N+Con组	100.03 ± 4.67	103.82 ± 14.08	3.75 ± 0.43	266.74 ± 26.08	5.11 ± 1.01	1.21 ± 0.02
N+H/R组	71.23 ± 4.23^a	275.96 ± 12.32^a	5.93 ± 0.19^a	86.61 ± 18.72^a	21.69 ± 0.37^a	0.54 ± 0.04^a
N+SPC组	86.66 ± 5.48^b	122.87 ± 8.03^b	6.94 ± 0.45^ab	172.88 ± 26.83^ab	10.92 ± 0.69^ab	0.78 ± 0.08^ab
H+Con组	91.15 ± 4.32^b	252.15 ± 24.19^abc	3.58 ± 0.33^bc	247.63 ± 21.45^abc	8.33 ± 0.07^abc	0.94 ± 0.02^ab
H+H/R组	65.74 ± 7.35^acd	367.52 ± 43.63^abcd	5.33 ± 0.22^abcd	38.59 ± 22.99^abcd	25.12 ± 0.46^abcd	0.46 ± 0.02^acd
H+SPC组	70.12 ± 3.81^acd	315.12 ± 25.22^abcde	5.36 ± 0.18^abcd	88.90 ± 17.33^acde	21.42 ± 2.11^acde	0.55 ± 0.11^acd
H+SPC+D组	83.72 ± 4.55^abef	154.33 ± 26.85^abdef	6.40 ± 0.42^abcdef	165.62 ± 24.28^abdef	15.43 ± 1.07^abcdef	0.82 ± 0.05^abef
H+SPC+D+A组	68.43 ± 3.20^acdg	264.96 ± 13.46^aceg	5.03 ± 0.12^abcdg	79.54 ± 19.22^acdeg	19.83 ± 0.23^acdeg	0.57 ± 0.05^acdg
F值	21.211	32.012	125.517	173.711	221.412	40.473
P值	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

分组	HK （U/mL）	PFK1 （U/mL）	PK （U/mL）
N+Con组	48.69 ± 4.00	79.38 ± 20.17	83.38 ± 8.63
N+H/R组	93.99 ± 4.59^a	148.81 ± 17.94^a	130.75 ± 3.18^a
N+SPC组	134.23 ± 8.84^ab	213.30 ± 12.36^ab	251.67 ± 14.45^ab
H+Con组	47.11 ± 4.51^bc	84.13 ± 13.24^bc	84.12 ± 7.71^bc
H+H/R组	57.71 ± 7.92^bcd	85.79 ± 12.67^bc	91.45 ± 5.03^bc
H+SPC组	72.43 ± 8.77^abcde	104.62 ± 10.65^abc	92.87 ± 5.14^bc
H+SPC+D组	122.34 ± 8.74^abcdef	192.45 ± 20.98^abdef	208.76 ± 32.65^abcdef
H+SPC+D+A组	66.83 ± 6.91^abcdg	77.83 ± 5.94^bcfg	132.80 ± 3.53^acdefg
F值	198.708	118.502	237.397
P值	< 0.001	< 0.001	< 0.001

分组	HK （U/mL）	PFK1 （U/mL）	PK （U/mL）
N+Con组	48.69 ± 4.00	79.38 ± 20.17	83.38 ± 8.63
N+H/R组	93.99 ± 4.59^a	148.81 ± 17.94^a	130.75 ± 3.18^a
N+SPC组	134.23 ± 8.84^ab	213.30 ± 12.36^ab	251.67 ± 14.45^ab
H+Con组	47.11 ± 4.51^bc	84.13 ± 13.24^bc	84.12 ± 7.71^bc
H+H/R组	57.71 ± 7.92^bcd	85.79 ± 12.67^bc	91.45 ± 5.03^bc
H+SPC组	72.43 ± 8.77^abcde	104.62 ± 10.65^abc	92.87 ± 5.14^bc
H+SPC+D组	122.34 ± 8.74^abcdef	192.45 ± 20.98^abdef	208.76 ± 32.65^abcdef
H+SPC+D+A组	66.83 ± 6.91^abcdg	77.83 ± 5.94^bcfg	132.80 ± 3.53^acdefg
F值	198.708	118.502	237.397
P值	< 0.001	< 0.001	< 0.001

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