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. 2020 Sep 10;20(1):274.
doi: 10.1186/s12906-020-03069-1.

Schisandra chinensis bee pollen's chemical profiles and protective effect against H2O2-induced apoptosis in H9c2 cardiomyocytes

Peiying Shi  1   2 , Qianqian Geng  1   2 , Lifu Chen  1   2 , Tianyu Du  1   2   3 , Yan Lin  1 , Rongcai Lai  1 , Fei Meng  1   2   3 , Zhenhong Wu  1   2 , Xiaoqing Miao  1   2 , Hong Yao  4
Affiliations

Schisandra chinensis bee pollen's chemical profiles and protective effect against H2O2-induced apoptosis in H9c2 cardiomyocytes

Peiying Shi et al. BMC Complement Med Ther. .

Abstract

Background: Schisandra chinensis (Turcz.) Baill bee pollen extract (SCBPE) is often used as a functional food in China due to its good antioxidant property. However, its chemical compositions and effects on H9c2 cardiomyocytes against H2O2-induced cell injury still lacks of reports thus far. This study aimed to characterize the main components of SCBPE and investigate its protective effects against H2O2-induced H9c2 cardiomyocyte injury.

Methods: The main components of SCBPE were analyzed via ultraperformance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF MS/MS). The three main nucleosides in SCBPE were quantitatively analyzed via ultraperformance liquid chromatography-diode array detection. Furthermore, the potential mechanism by which SCBPE exerts protective effects against H2O2-induced H9c2 cardiomyocyte injury was explored for the first time via cell survival rate measurements; cell morphological observation; myocardial superoxide dismutase (SOD) activity and malondialdehyde (MDA) and glutathione (GSH) level determination; flow cytometry; and quantitative polymerase chain reaction.

Results: Two carbohydrates, three nucleosides, and nine quinic acid nitrogen-containing derivatives in SCBPE were identified or tentatively characterized via UPLC-QTOF MS/MS. The nine quinic acid nitrogen-containing derivatives were first reported in bee pollen. The contents of uridine, guanosine, and adenosine were 2.4945 ± 0.0185, 0.1896 ± 0.0049, and 1.8418 ± 0.0157 μg/mg, respectively. Results of in vitro experiments showed that cell survival rate, myocardial SOD activity, and GSH level significantly increased and myocardial MDA level significantly decreased in SCBPE groups compared with those in H2O2 group. Cell morphology in SCBPE groups also markedly improved compared with that in H2O2 group. Results indicated that SCBPE protected H9c2 cardiomyocytes from H2O2-induced apoptosis by downregulating the mRNA expressions of Bax, cytochrome C, and caspase-3 and upregulating the Bcl-2 mRNA expression.

Conclusions: This study is the first to report that SCBPE could protect against oxidative stress injury and apoptosis in H2O2-injured H9c2 cells. Results indicated that the nucleosides and quinic acid nitrogen-containing derivatives could be the main substances that exert protective effects against H2O2-induced H9c2 cardiomyocyte injury.

Keywords: H2O2; H9c2 cardiomyocytes; Nucleosides; Protective effect; Quinic acid nitrogen-containing derivatives; Schisandra chinensis bee pollen extract.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
UV chromatogram at 254 nm (a), TIC chromatograms in positive ion mode (b) and in negative ion mode (c) of SCBPE
Fig. 2
Fig. 2
The proposed (−)ESI-Q-TOF MS/MS fragmentation pathway of 3-caffeoyl-5-feruloylquinic acid nitrogen-containing derivative
Fig. 3
Fig. 3
The proposed (−)ESI-Q-TOF MS/MS fragmentation pathway of 3-caffeoyl-5-3′-methoxycinnamylquinic acid nitrogen-containing derivative
Fig. 4
Fig. 4
The effects of SCBPE on morphology of H9c2 myocardial cells. a, negative control group; b, H2O2 group; c, positive control (Vc) group; d, 12.5 μg/mL SCBPE; e, 25 μg/mL SCBPE; f, 50 μg/mL SCBPE
Fig. 5
Fig. 5
Effect of SCBPE on apoptosis of H9c2 cardiomyocytes induced by H2O2. C07, negative control group; C08, H2O2 group; C09, positive control (Vc) group; C10, 12.5 μg/mL SCBPE; C11, 25 μg/mL SCBPE; C12, 50 μg/mL SCBPE. * Compared with H2O2 group, p < 0.01
Fig. 6
Fig. 6
The effect of SCBPE on the mRNA expression of caspase-3, cytochrome C, Bcl-2, Bax, and Bcl-2/Bax induced by H2O2 in H9c2 myocardial cells of each group. G1, negative control group; G2, H2O2 group; G3, positive control (Vc) group; G4, 12.5 μg/mL SCBPE; G5, 25 μg/mL SCBPE; G6, 50 μg/mL SCBPE. Compared with H2O2 group, # p < 0.05, * p < 0.01
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