The protective mechanism of differential isomers of glycyrrhetinic acid on cisplatin-induced H9c2 cardiomyocyte injury

doi:10.11958/20220987

Abstract

Abstract:

Objective To explore the protective mechanism of 18α-glycyrrhetinic acid (GA) and 18β-GA, the differential isomers of glycyrrhetinic acid, on cisplatin (CDDP) -induced H9c2 cardiomyocyte injury. Methods CCK-8 assay was used to detect cell viability and the concentration of CDDP-induced cell injury. The safe doses of 18α-GA and 18β-GA, and the effective concentrations of 18α-GA and 18β-GA to improve the decrease of cardiomyocyte viability induced by CDDP were screened. The cells were divided into the control group, the CDDP group, the 18α-GA group and the 18β-GA group. Hoechst staining was used to detect apoptosis in each group. The level of reactive oxygen species (ROS) was detected by DCFH-DA. Mito-Tracker Red CMXRos staining was used to assess mitochondrial activity. The expressions of C-Caspase3, B-lymphocytoma gene-2 (Bcl-2), Bcl-2 associated X protein (Bax) and cytochrome C (Cyt-c) were detected by Western blot assay in each group. Results The results of CCK-8 assay showed that CDDP at 20 μmol/L significantly decreased the viability of H9c2 cardiomyocytes (P<0.01). When the concentrations of 18α-GA and 18β-GA were less than 100 μmol/L, there were no significant effects on cardiomyocytes, and 50 and 100 μmol/L could improve the decrease of cardiomyocyte viability caused by CDDP (P<0.01). Compared with the control group, the apoptosis was obvious in the CDDP group, the ROS level was increased, the mitochondrial foot length and mitochondria with reticular structure were decreased, the protein expression levels of C-Caspase3 and Cyt-c were increased, and the protein expression of Bcl-2/Bax was decreased (P<0.05). Compared with the CDDP group, cell apoptosis was improved in the 18α-GA group and the 18β-GA group, ROS level decreased, mitochondrial foot length and mitochondria with reticular structure increased, protein expression levels of C-Caspase3 and Cyt-c decreased, and protein expression levels of Bcl-2/Bax increased (P<0.05). Conclusion 18α-GA and 18β-GA can reduce the apoptosis of H9c2 cardiomyocytes induced by CDDP by inhibiting ROS level and protecting mitochondrial function.

Key words: cisplatin, cardiotoxicity, glycyrrhetinic acid, reactive oxygen, mitochondria, species differential isomers

CLC Number:

R285.5

Figures/Tables 9

Fig.1 Effects of CDDP on viability of H9c2 cardiomyocytes

Fig.2 Effects of 18α-GA and 18β-GA on the viability of H9c2 cells

Fig.3 Effects of low, medium and high doses of 18α-GA and 18β-GA on cisplatin-induced myocardial cell viability

Fig.4 Effects of 18α-GA and 18β-GA on CDDP-induced apoptosis of cardiomyocytes (Hoechst staining, ×400)

Fig.5 Effects of 18α-GA and 18β-GA on ROS content in cardiomyocytes induced by cisplatin

Fig.6 Effects of 18α-GA and 18β-GA on mitochondrial activity of cardiomyocytes induced by CDDP (×400)

Tab.1 Comparison of mitochondrial foot length and number of mitochondria with reticular structure between the four groups

组别	线粒体足长度	具有网状结构的线粒体数量
对照组	1.00±0.20	0.94（0.75，1.22）
CDDP组	0.53±0.03^a	0.38（0.38，0.47）^a
18α-GA组	0.79±0.07^b	0.75（0.75，0.84）^b
18β-GA组	0.87±0.21^b	0.75（0.75，0.75）^b
F或H	34.748^＊＊	13.320^＊＊

Tab.2 Comparison of protein expression levels of C-Caspase3, Bcl-2/Bax and Cyt-c between the four groups

组别	C-Caspase3	Bcl-2/Bax	Cyt-c
对照组	1.06±0.40	1.00±0.15	0.97±0.41
CDDP组	2.15±0.40^a	0.50±0.04^a	2.66±0.22^a
18α-GA组	1.30±0.28^b	0.97±0.18^b	1.11±0.25^b
18β-GA组	1.30±0.35^b	0.88±0.09^b	0.57±0.04^b
F	10.321^＊＊	9.768^＊＊	36.039^＊＊

Fig.7 Effects of 18α-GA and 18β-GA on CDDP-induced mitochondrial-related apoptostic protein expression in cardiomyocytes

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