Effect of Baicalein on UVB-induced HaCaT Cell Light Aging and p38 Signaling Pathway

YANG Liu; WANG Ye-qiu; ZHANG Ning; LI Jian-min

doi:10.13422/j.cnki.syfjx.2017160139

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Effect of Baicalein on UVB-induced HaCaT Cell Light Aging and p38 Signaling Pathway

更新时间：2024-01-04

- Effect of Baicalein on UVB-induced HaCaT Cell Light Aging and p38 Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 16, Pages: 139-144(2017)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.2017160139
  CLC： R285
- Published：2017
- 稿件说明：
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YANG Liu, WANG Ye-qiu, ZHANG Ning, et al. Effect of Baicalein on UVB-induced HaCaT Cell Light Aging and p38 Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(16): 139-144.
DOI：

YANG Liu, WANG Ye-qiu, ZHANG Ning, et al. Effect of Baicalein on UVB-induced HaCaT Cell Light Aging and p38 Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(16): 139-144. DOI： 10.13422/j.cnki.syfjx.2017160139.

摘要

目的：研究黄芩素对中波紫外线（UVB）诱导人皮肤角质形成细胞（HaCaT）光老化及相关p38丝裂原活化蛋白激酶（p38MAPK）信号通路的影响。方法：选择1×10-11，1×10-10，1×10-9，1×10-8，1×10-7，1×10-6，1×10-5，1×10-4，1×10-3 mol·L-19种浓度黄芩素作用于HaCaT细胞，噻唑蓝（MTT）法筛选出最佳有效浓度。经预实验筛选出辐射强度为0.5 mW·cm-2的UVB，辐射时间为5 min建立光老化模型，筛选出最佳有效浓度作用于光老化模型，另设空白组MTT法检测各组细胞活性。超氧化物歧化酶（SOD），谷胱甘肽过氧化物酶（GSH-Px），过氧化氢酶（CAT）及丙二醛（MDA）试剂盒检测SOD，GSH-Px，CAT活性及MDA含量。实时定量荧光定量聚合酶连式反应（RT-PCR）及蛋白免疫印迹法（Western blot）分别检测各组细胞中mRNA及蛋白表达。结果：筛选出1×10-7，1×10-6，1×10-5 mol·L-1黄芩素为最佳有效浓度；与空白组比较，UVB组对HaCaT细胞无明显的增殖作用。与UVB组比较，1×10-7，1×10-6，1×10-5 mol·L-1黄芩素组对光老化模型无明显增殖作用。与空白组比较，UVB组SOD，GSH-Px及CAT活性明显的降低，MDA含量显著升高（P <0.01）；与UVB组比较，1×10-7，1×10-6，1×10-5 mol·L-1黄芩素组SOD，GSH，CAT活性明显升高，MDA含量明显降低（P <0.05，P <0.01）。与空白组比较，UVB组肿瘤坏死因子-α （TNF-α）mRNA及TNF-α，磷酸化p38（p-p38）蛋白表达显著升高（P <0.01）；与UVB组比较，1×10-7，1×10-6，1×10-5 mol·L-1黄芩素组TNF-α mRNA及TNF-α，p-p38蛋白表达明显降低（P <0.05，P <0.01）。结论：黄芩素对UVB诱导HaCaT细胞光老化保护作用主要是通过提高氧化酶活性以及阻断p38MAPK信号通路，抑制炎症因子产生。

Abstract

Objective: To study the effect of baicalein on ultraviolet B(UVB) induced-human skin keratinocyte (HaCaT) cell light aging and p38 mitogen activated protein kinase (p38MAPK) signaling pathways. Method: The 1×10-11

1×10-10

1×10-9

1×10-8

1×10-7

1×10-6

1×10-5

1×10-4

1×10-3 mol·L-1 baicalein were used on HaCaT cells

and the best effective concentration was determined by methyl thiazolyl tetrazolium(MTT) method. In the preliminary experiments

UVB with radiation intensity of 0.5 mW·cm-2 was selected

and with radiation time for 5 min to establish light aging models. Then the best effective concentration was used on the light aging models. MTT method was used to detect the cells activity in each group; superoxide dismutase(SOD)

glutathione peroxidase(GSH-Px)

catalase(CAT) and malondialdehyde(MDA) kits were used to detect SOD

GSH-Px

CAT activity and MDA content respectively. Real-time quantitative PCR and Western blot were used respectively to detect mRNA and protein expression levels in each group. Result: 1×10-7

1×10-6

1×10-5 mol·L-1 baicalein were selected as the best effective concentrations. As compared with the blank group

UVB had no significant proliferation effect on HaCaT cells. As compared with UVB group

1×10-7

1×10-6

and 1×10-5mol·L-1 baicalein had no obvious effect on the proliferation of light aging models. As compared with the blank group

SOD

GSH-Px and CAT activities were significantly decreased while MDA contend was increased in UVB group (P <0.01). As compared with UVB model group

1×10-7

1×10-6

1×10-5 mol·L-1 baicalein could increase SOD

GSH-Px and CAT activities

and decrease MDA contend (P <0.05

P <0.01). As compared with the blank group

the expression levels of tumor necrosis factor (TNF)-α mRNA as well as TNF-α and p38 protein were significantly increased in UVB group (P <0.01). As compared with UVB model group

1×10-7

1×10-6

1×10-5 mol·L-1 baicalein could decrease the expression levels of TNF-α mRNA as well as TNF-α and p38 protein (P <0.05

P <0.01). Conclusion: Baicalein has the protective effect on UVB-induced cell light aging

and its mechanism is mainly associated with tenhancing GSH-Px

SOD and CAT activities

blocking p38MAPK signaling pathway

and inhibiting inflammatory cytokines.

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