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Effects of ginsenoside Rg2 on the ultraviolet B-induced DNA damage responses in HaCaT cells

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Abstract

Our previous study demonstrated the increase in the repair of UVB damage by mRg2, a mixture of ginsenosides containing 60% Rg2 in NIH3T3 cells. In the present study, the effects of purified Rg2 on the repair and apoptosis in ultraviolet B (UVB)-exposed HaCaT cells were investigated on gene expression levels. When cells were exposed to UVB and post-incubated in normal medium for 24 h, the cell viability decreased to about 50% of that in nontreated control. When Rg2 was post-incubated, however, the UVB-induced cytotoxicity was significantly prevented in an Rg2 concentration- and time-dependent manner. The apoptotic nuclear fragmentation resulting from UVB exposure was also significantly protected by the Rg2 post-incubation. Microarray analysis showed that the genes stimulated by the Rg2-alone treatment include those involved in p53 signaling pathway such as GADD45α, GADD45β, and cell communication genes. RT-PCR analysis showed that the Rg2-alone treatment slightly upregulated the p53 and GADD45 transcript and protein levels by about 1.5-fold as compared with the nontreated control. The mRNA levels of p53 and GADD45 in cells exposed to UVB and post-incubated with Rg2 for 24 h decreased in an Rg2 concentration-dependent manner as compared with that post-incubated in normal medium. However, the mRNA level of the UVB-exposed cells post-incubated with 5 μM retinol was essentially the same as that post-incubated in normal medium. Time course experiment showed that the mRNA levels of p53 and GADD45 in UVB-exposed cells were upregulated by post-incubation with 50 μM Rg2 until 6 and 9 h, respectively, and then gradually decreased until 24 h. By Western blot analysis, it was also revealed that the Rg2 post-incubation decreases the expression of p53, phospho-p53, GADD45, and ATM in UVB-exposed cells. Time course analysis also indicated that these decreased expressions were due to the earlier upregulation of p53 and GADD45 proteins. When UVB-exposed cells were post-incubated with Rg2 for 24 h after UVB exposure, the level of remaining cyclobutane pyrimidine dimers decreased in both Rg2 concentration- and time-dependent manner. All these results suggest that Rg2 protects cells against UVB-induced genotoxicity by increasing DNA repair, in possible association with modulation of protein levels involved in p53 signaling pathway.

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Acknowledgement

The present study was supported by SMBA (2008-JB-SMBA). Dae Hyun Shin, Hack Soo Kim, and Bo Hyeon Kim were supported by Somang Cosmetics, Korea.

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Authors and Affiliations

  1. Division of Biological Science, Institute of Natural Sciences, Wonkwang University, Iksan, Chonbuk, 570-749, South Korea

    Se Eun Ha, Hyung Do Kim, Sun Mi Shim & Jong Kun Park

  2. Somang Cosmetics R&D Center, Incheon, South Korea

    Dae Hyun Shin, Hack Soo Kim & Bo Hyeon Kim

  3. Department of Biotechnology, Chosun University, Kwangju, South Korea

    Jung Sup Lee

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  1. Se Eun Ha
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Correspondence to Jong Kun Park.

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Ha, S.E., Shin, D.H., Kim, H.D. et al. Effects of ginsenoside Rg2 on the ultraviolet B-induced DNA damage responses in HaCaT cells. Naunyn-Schmied Arch Pharmacol 382, 89–101 (2010). https://doi.org/10.1007/s00210-010-0522-9

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