Effect of Salvia miltiorrhiza on aflatoxin B1-induced oxidative stress in cultured rat hepatocytes

Jin Liu; Cheng Feng Yang; Bee Lan Lee; Han Ming Shen; Siau Gek Ang; Choon Nam Ong

doi:10.1080/10715769900301131

Effect of Salvia miltiorrhiza on aflatoxin B₁-induced oxidative stress in cultured rat hepatocytes

Jin Liu, Cheng Feng Yang, Bee Lan Lee, Han Ming Shen, Siau Gek Ang, Choon Nam Ong

Markey Cancer Center / Cancer Research Priority Initiative

Research output: Contribution to journal › Article › peer-review

41 Scopus citations

Abstract

Recent findings have suggested that oxidative damage might contribute to the cytotoxicity and carcinogenicity of aflatoxin B₁ (AFB₁). Salvia miltiorrhiza (Sm), a herbal plant that has been used extensively in traditional Chinese medicine for treating cardiovascular and liver diseases, is believed to have some antioxidative capabilities. In this study, the protective effect of Sm against AFB₁-induced cytotoxicity was investigated in cultured primary rat hepatocytes. AFB₁-induced cytotoxicity and lipid peroxidation (LPO) were estimated by determination of lactate dehydrogenase (LDH) leakage and thiobarbituric acid reactive substances (TBARS) formation, respectively. Intracellular reactive oxygen species (ROS) formation was measured using a fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH- DA). In addition, changes of intracellular glutathione (GSH) content were also studied. Results showed that Sm was able to suppress the LDH leakage induced by AFB₁ in a dose-dependent manner. A dose-dependent inhibitory effect of Sm on AFB₁-induced LPO was also found in hepatocytes treated with Sm. It was further observed that Sm produced an inhibitory effect on ROS formation caused by AFB₁. Concomitantly, the GSH content in Sm-treated groups increased substantially compared to those without Sm treatment. These findings suggest that Sm can inhibit the cytotoxicity of AFB₁ through decreasing ROS formation, inhibiting LPO and preventing GSH depletion. The major component of the aqueous extract of Sm was identified by using high performance liquid chromatography (HPLC), proton magnetic resonance (¹H-NMR) and mass spectrum (MS). Analytical results suggested that D(+)β3,4- dihydroxyphenol lactic acid (DA) is the main compound of the aqueous extract of Sm.

Original language	English
Pages (from-to)	559-568
Number of pages	10
Journal	Free Radical Research
Volume	31
Issue number	6
DOIs	https://doi.org/10.1080/10715769900301131
State	Published - 1999

Bibliographical note

Funding Information:
The authors would like to thank Mr. H.Y. Ong and Ms. X. Wang for their technical assistance. Liu, J. and Yang, C.E are supported by research scholarship from National University of Singapore. This work was partly supported by a grant from the National Medical Research Council of Singapore (RP3970301N).

Funding

The authors would like to thank Mr. H.Y. Ong and Ms. X. Wang for their technical assistance. Liu, J. and Yang, C.E are supported by research scholarship from National University of Singapore. This work was partly supported by a grant from the National Medical Research Council of Singapore (RP3970301N).

Funders	Funder number
National Health and Medical Research Council Clinical Trials Centre	RP3970301N
National University Hospital, Singapore

Keywords

4-dihydroxyphenol lactic acid
Aflatoxin B
Cytotoxicity
D(+)β3
Primary hepatocyte
Reactive oxygen species
Salvia miltiorrhiza

ASJC Scopus subject areas

Biochemistry

Access to Document

10.1080/10715769900301131

Cite this

@article{294fa59f968e431489263ec245011a16,

title = "Effect of Salvia miltiorrhiza on aflatoxin B1-induced oxidative stress in cultured rat hepatocytes",

abstract = "Recent findings have suggested that oxidative damage might contribute to the cytotoxicity and carcinogenicity of aflatoxin B1 (AFB1). Salvia miltiorrhiza (Sm), a herbal plant that has been used extensively in traditional Chinese medicine for treating cardiovascular and liver diseases, is believed to have some antioxidative capabilities. In this study, the protective effect of Sm against AFB1-induced cytotoxicity was investigated in cultured primary rat hepatocytes. AFB1-induced cytotoxicity and lipid peroxidation (LPO) were estimated by determination of lactate dehydrogenase (LDH) leakage and thiobarbituric acid reactive substances (TBARS) formation, respectively. Intracellular reactive oxygen species (ROS) formation was measured using a fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH- DA). In addition, changes of intracellular glutathione (GSH) content were also studied. Results showed that Sm was able to suppress the LDH leakage induced by AFB1 in a dose-dependent manner. A dose-dependent inhibitory effect of Sm on AFB1-induced LPO was also found in hepatocytes treated with Sm. It was further observed that Sm produced an inhibitory effect on ROS formation caused by AFB1. Concomitantly, the GSH content in Sm-treated groups increased substantially compared to those without Sm treatment. These findings suggest that Sm can inhibit the cytotoxicity of AFB1 through decreasing ROS formation, inhibiting LPO and preventing GSH depletion. The major component of the aqueous extract of Sm was identified by using high performance liquid chromatography (HPLC), proton magnetic resonance (1H-NMR) and mass spectrum (MS). Analytical results suggested that D(+)β3,4- dihydroxyphenol lactic acid (DA) is the main compound of the aqueous extract of Sm.",

keywords = "4-dihydroxyphenol lactic acid, Aflatoxin B, Cytotoxicity, D(+)β3, Primary hepatocyte, Reactive oxygen species, Salvia miltiorrhiza",

author = "Jin Liu and Yang, {Cheng Feng} and Lee, {Bee Lan} and Shen, {Han Ming} and Ang, {Siau Gek} and Ong, {Choon Nam}",

note = "Funding Information: The authors would like to thank Mr. H.Y. Ong and Ms. X. Wang for their technical assistance. Liu, J. and Yang, C.E are supported by research scholarship from National University of Singapore. This work was partly supported by a grant from the National Medical Research Council of Singapore (RP3970301N).",

year = "1999",

doi = "10.1080/10715769900301131",

language = "English",

volume = "31",

pages = "559--568",

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TY - JOUR

T1 - Effect of Salvia miltiorrhiza on aflatoxin B1-induced oxidative stress in cultured rat hepatocytes

AU - Liu, Jin

AU - Yang, Cheng Feng

AU - Lee, Bee Lan

AU - Shen, Han Ming

AU - Ang, Siau Gek

AU - Ong, Choon Nam

N1 - Funding Information: The authors would like to thank Mr. H.Y. Ong and Ms. X. Wang for their technical assistance. Liu, J. and Yang, C.E are supported by research scholarship from National University of Singapore. This work was partly supported by a grant from the National Medical Research Council of Singapore (RP3970301N).

PY - 1999

Y1 - 1999

N2 - Recent findings have suggested that oxidative damage might contribute to the cytotoxicity and carcinogenicity of aflatoxin B1 (AFB1). Salvia miltiorrhiza (Sm), a herbal plant that has been used extensively in traditional Chinese medicine for treating cardiovascular and liver diseases, is believed to have some antioxidative capabilities. In this study, the protective effect of Sm against AFB1-induced cytotoxicity was investigated in cultured primary rat hepatocytes. AFB1-induced cytotoxicity and lipid peroxidation (LPO) were estimated by determination of lactate dehydrogenase (LDH) leakage and thiobarbituric acid reactive substances (TBARS) formation, respectively. Intracellular reactive oxygen species (ROS) formation was measured using a fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH- DA). In addition, changes of intracellular glutathione (GSH) content were also studied. Results showed that Sm was able to suppress the LDH leakage induced by AFB1 in a dose-dependent manner. A dose-dependent inhibitory effect of Sm on AFB1-induced LPO was also found in hepatocytes treated with Sm. It was further observed that Sm produced an inhibitory effect on ROS formation caused by AFB1. Concomitantly, the GSH content in Sm-treated groups increased substantially compared to those without Sm treatment. These findings suggest that Sm can inhibit the cytotoxicity of AFB1 through decreasing ROS formation, inhibiting LPO and preventing GSH depletion. The major component of the aqueous extract of Sm was identified by using high performance liquid chromatography (HPLC), proton magnetic resonance (1H-NMR) and mass spectrum (MS). Analytical results suggested that D(+)β3,4- dihydroxyphenol lactic acid (DA) is the main compound of the aqueous extract of Sm.

AB - Recent findings have suggested that oxidative damage might contribute to the cytotoxicity and carcinogenicity of aflatoxin B1 (AFB1). Salvia miltiorrhiza (Sm), a herbal plant that has been used extensively in traditional Chinese medicine for treating cardiovascular and liver diseases, is believed to have some antioxidative capabilities. In this study, the protective effect of Sm against AFB1-induced cytotoxicity was investigated in cultured primary rat hepatocytes. AFB1-induced cytotoxicity and lipid peroxidation (LPO) were estimated by determination of lactate dehydrogenase (LDH) leakage and thiobarbituric acid reactive substances (TBARS) formation, respectively. Intracellular reactive oxygen species (ROS) formation was measured using a fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH- DA). In addition, changes of intracellular glutathione (GSH) content were also studied. Results showed that Sm was able to suppress the LDH leakage induced by AFB1 in a dose-dependent manner. A dose-dependent inhibitory effect of Sm on AFB1-induced LPO was also found in hepatocytes treated with Sm. It was further observed that Sm produced an inhibitory effect on ROS formation caused by AFB1. Concomitantly, the GSH content in Sm-treated groups increased substantially compared to those without Sm treatment. These findings suggest that Sm can inhibit the cytotoxicity of AFB1 through decreasing ROS formation, inhibiting LPO and preventing GSH depletion. The major component of the aqueous extract of Sm was identified by using high performance liquid chromatography (HPLC), proton magnetic resonance (1H-NMR) and mass spectrum (MS). Analytical results suggested that D(+)β3,4- dihydroxyphenol lactic acid (DA) is the main compound of the aqueous extract of Sm.

KW - 4-dihydroxyphenol lactic acid

KW - Aflatoxin B

KW - Cytotoxicity

KW - D(+)β3

KW - Primary hepatocyte

KW - Reactive oxygen species

KW - Salvia miltiorrhiza

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