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Crude polysaccharide from an anti-UVB cell clone of Bupleurum scorzonerifolium protect HaCaT cells against UVB-induced oxidative stress

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Abstract

Bupleurum scorzonerifolium Willd has been found to have a wide range of immunopharmacologic functions. We isolated an anti-UVB B. scorzonerifolium cell clone and found elevated level of polysaccharides. In this study, we investigated the ability of crude polysaccharide (CP) from the anti-UVB B. scorzonerifolium cell clone to inhibit UVB-induced photodamage using a human skin keratinocyte cell line, HaCaT. Cells were UVB irradiated and then incubated in presence of different concentrations of CP. MTT assay showed that the CP did not induce cytotoxic effect under 10 mg/mL and after UVB irradiation, CP can inhibit UVB-induced HaCaT cell death. Decreased reactive oxygen species and lipid peroxidation and increased superoxide dismutase activity showed that CP can act as a free radical scavenger. Furthermore, CP had a strong protective ability against UVB-induced DNA damage. These effects were compared to the crude polysaccharide (CP′) from normal B. scorzonerifolium callus at concentration of 20 mg/mL. The portion of crude polysaccharide (CP) from the anti-UVB B. scorzonerifolium cell clone was more than 2.5-fold higher than crude polysaccharide (CP′) from normal B. scorzonerifolium callus. Taken together, the protective mechanisms of crude polysaccharide from the anti-UVB B. scorzonerifolium cell clone against UVB-induced photodamage occur by the inhibition of UVB-induced reactive oxygen species production, lipid peroxidation and DNA damage.

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Abbreviations

2,4-D:

2,4-dichlorophenoxyaceic acid

CM-DCF:

Chloromethyl 2′,7′-dichlorodihydrofluorescein diacetate

CM-H2DCFDA:

5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate acetyl ester

CP:

Crude polysaccharides from anti-UVB B. scorzonerifolium cell clone

CP′:

Crude polysaccharides from normal callus of B. scorzonerifolium

CP*:

Crude polysaccharides from the aerial parts of Wild B. scorzonerifolium

DMEM:

Dulbecco’s modified Eagle medium

DMSO:

Dimethylsulfoxide

FCS:

Fetal calf serum

MDA:

Malondialdehyde

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

UVB:

Ultraviolet-B

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (30760089).

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Authors and Affiliations

  1. Plant Science Institute, School of Life Sciences, Yunnan University, Kunming, 650091, China

    Jinran Dai, Haiyin Ma, Jing Fan, Yuzhong Li, Jianguang Wang, Hongmei Ni & Suiyun Chen

  2. School of Life Sciences, Shandong University, Jinan, 250100, China

    Guangmin Xia

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  1. Jinran Dai
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  2. Haiyin Ma
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  3. Jing Fan
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  4. Yuzhong Li
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  5. Jianguang Wang
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  6. Hongmei Ni
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  7. Guangmin Xia
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  8. Suiyun Chen
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Correspondence to Suiyun Chen.

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Jinran Dai and Haiyin Ma authors contribute equally to this article.

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Dai, J., Ma, H., Fan, J. et al. Crude polysaccharide from an anti-UVB cell clone of Bupleurum scorzonerifolium protect HaCaT cells against UVB-induced oxidative stress. Cytotechnology 63, 599–607 (2011). https://doi.org/10.1007/s10616-011-9381-6

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