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. 2015 Dec 4;10(12):e0143528.
doi: 10.1371/journal.pone.0143528. eCollection 2015.

Aloin Protects Skin Fibroblasts From Heat Stress-Induced Oxidative Stress Damage by Regulating the Oxidative Defense System

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Free PMC article

Aloin Protects Skin Fibroblasts From Heat Stress-Induced Oxidative Stress Damage by Regulating the Oxidative Defense System

Fu-Wei Liu et al. PLoS One. .
Free PMC article

Abstract

Oxidative stress is commonly involved in the pathogenesis of skin damage induced by environmental factors, such as heat stress. Skin fibroblasts are responsible for the connective tissue regeneration and the skin recovery from injury. Aloin, a bioactive compound in Aloe vera, has been reported to have various pharmacological activities, such as anti-inflammatory effects. The aim of this study was to investigate the protective effect of aloin against heat stress-mediated oxidative stress in human skin fibroblast Hs68 cells. Hs68 cells were first incubated at 43°C for 30 min to mimic heat stress. The study was further examined if aloin has any effect on heat stress-induced oxidative stress. We found that aloin protected Hs68 cells against heat stress-induced damage, as assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assay. Aloin protected Hs68 cells by regulating reactive oxygen species production and increasing the levels of glutathione, cytosolic and mitochondrial superoxide dismutase. Aloin also prevented the elevation of thiobarbituric acid reactive substances and the reduction of 8-OH-dG induced by heat stress. These results indicated that aloin protected human skin fibroblasts from heat stress-induced oxidative stress damage by regulating the oxidative defense system.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Effect of Aloin on cell viability of human foreskin fibroblast Hs68 cells exposed to 43°Cwith/without Aloin supplement.
(A) The optical microscopy images (magnification, ×100) of fibroblast cell and (B) diagram of curve of cell viabilities were representative of three independent experiments. Cells were exposed to heat stress with a single administration of Aloin (43°C+A150) or (43°C+A300), vehicle (43°C), or were left unheated and given the vehicle (CT). The cell viabilities were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The data are expressed as the means ± SEM of two replicates of three samples (n = 6). The asterisks indicate a significant difference (* P<0.05, ** P<0.01, *** P<0.005) compared to the control group; the crosses indicate a significant difference (++ P<0.01, +++ P<0.005) compared to the heat-treated group.
Fig 2
Fig 2. Effect of Aloin on lactate dehydrogenase (LDH) release in Hs68 cells with/without heat stress challenge.
Cells were exposed to heat stress with a single administration of Aloin (43°C+A150) or (43°C+A300), vehicle (43°C), or were left unheated and given the vehicle (CT). The LDH activities were measured by ELISA. The data are expressed as the means ± SEM of two replicates of three samples (n = 6). The asterisks indicate a significant difference (*** P<0.005) compared to the control group; the crosses indicate a significant difference (+++ P<0.005) compared to the heat-treated group.
Fig 3
Fig 3. Effect of Aloin on intracellular ROS release in Hs68 cells with/without heat stress challenge.
Cells were exposed to heat stress with a single administration of Aloin (43°C+A150) or (43°C+A300), vehicle (43°C), or were left unheated and given the vehicle (CT). Representative staining of ROS in HS68 cultures probed with DHR-123 by flow cytometry analysis (A). The data are expressed as the means ± SEM of two replicates of three samples (n = 6) (B). The asterisks indicate a significant difference (* P<0.05, *** P<0.005) compared to the control group; the crosses indicate a significant difference (++ P<0.01, +++ P<0.005) compared to the heat-treated group. Experiments were performed in duplicates with Hs68 cultures derived from 5 different donors.
Fig 4
Fig 4. Effect of Aloin on 8-OH-dG concentration in Hs68 cells with/without heat stress challenge.
Cells were exposed to heat stress with a single administration of Aloin (43°C+A150) or (43°C+A300), vehicle (43°C), or were left unheated and given the vehicle (CT). The 8-OH-dG levels were measured by ELISA. The data are expressed as the means ± SEM of two replicates of three samples (n = 6). The asterisks indicate a significant difference (* P<0.05) compared to the control group; the crosses indicate a significant difference (+ P<0.05) compared to the heat-treated group.
Fig 5
Fig 5. Effect of Aloin on total GSH levels in Hs68 cells with/without heat stress challenge.
Cells were exposed to heat stress with a single administration of Aloin (43°C+A150) or (43°C+A300), vehicle (43°C), or were left unheated and given the vehicle (CT). The GSH levels were measured by ELISA. The data are expressed as the means ± SEM of two replicates of three samples (n = 6). The asterisks indicate a significant difference (* P<0.05, ** P<0.01, *** P<0.005) compared to the control group; the crosses indicate a significant difference (+ P<0.05, ++ P<0.01, +++ P<0.005) compared to the heat-treated group.
Fig 6
Fig 6. Effect of Aloin on (A) cytosolic and (B) mitochondrial SOD activities in Hs68 cells with/without heat stress challenge.
Cells were exposed to heat stress with a single administration of Aloin (43°C+A150) or (43°C+A300), vehicle (43°C), or were left unheated and given the vehicle (CT). The cytosolic and mitochondrial SOD activities were assayed by ELISA. The data are expressed as the means ± SEM of two replicates of three samples (n = 6). The asterisks indicate a significant difference (* P<0.05, *** P<0.005) compared to the control group; the crosses indicate a significant difference (++ P<0.01, +++ P<0.005) compared to the heat-treated group.

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Grant support

This work was partially supported by grants from the National Science Council (NSC102-2314-B-182A-051-MY3) and Chang Gung Memorial Hospital (CMRPG3B1053 and CMRPG3E1551) to Huang-Ping Yu. Support was also provided by the National Science Council (NSC103-2314-B-182-046-MY2) and Chang Gung Memorial Hospital (CMRPG3E1541) to Fu-Chao Liu. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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