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Role for heat shock protein 90α in the proliferation and migration of HaCaT cells and in the deep second-degree burn wound healing in mice.

Zhang Y, Bai X, Wang Y, Li N, Li X, Han F, Su L, Hu D - PLoS ONE (2014)

Bottom Line: The hypoxic wound microenvironment promotes cell migration through a hypoxia--heat shock protein 90 alpha (Hsp90α)--low density lipoprotein receptor-related protein-1 (LRP-1) autocrine loop.Experiments performed with a human keratinocyte cell line--HaCaT also confirmed above results. 17-dimethylaminoethylamino-17demethoxygeldanamycin hydrochloride (17-DMAG), an Hsp90α inhibitor, was used to further evaluate the function of Hsp90α in wound healing.In conclusion, these results provided a rationale for the therapeutic effect of Hsp90α on the burn wound management.

View Article: PubMed Central - PubMed

Affiliation: Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shannxi, China.

ABSTRACT
Inflammation, proliferation, and tissue remodeling are essential steps for wound healing. The hypoxic wound microenvironment promotes cell migration through a hypoxia--heat shock protein 90 alpha (Hsp90α)--low density lipoprotein receptor-related protein-1 (LRP-1) autocrine loop. To elucidate the role of this autocrine loop on burn wound healing, we investigated the expression profile of Hsp90α at the edge of burn wounds and found a transient increase in both mRNA and protein levels. Experiments performed with a human keratinocyte cell line--HaCaT also confirmed above results. 17-dimethylaminoethylamino-17demethoxygeldanamycin hydrochloride (17-DMAG), an Hsp90α inhibitor, was used to further evaluate the function of Hsp90α in wound healing. Consistently, topical application of Hsp90α in the early stage of deep second-degree burn wounds led to reduced inflammation and increased tissue granulation, with a concomitant reduction in the size of the wound at each time point tested (p<0.05). Consequently, epidermal cells at the wound margin progressed more rapidly causing an expedited healing process. In conclusion, these results provided a rationale for the therapeutic effect of Hsp90α on the burn wound management.

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IHC assay showing the changes of Hsp90α immunostaining in burned mouse skin.(A) Unwound normal skin showed a few positive Hsp90α stainings. (C, E) Epidermal and dermal tissues after burn injury appeared more positive brownish stainings, indicating that Hsp90α was induced after the burn stimulation. In addition, Hsp90α level was the highest at 12 h post-treatment (C) and somewhat decreased at 48 h (E). Magnification of red boxed areas in (A), (C) and (E) was shown as (B), (D)and (F), respectively.
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pone-0103723-g003: IHC assay showing the changes of Hsp90α immunostaining in burned mouse skin.(A) Unwound normal skin showed a few positive Hsp90α stainings. (C, E) Epidermal and dermal tissues after burn injury appeared more positive brownish stainings, indicating that Hsp90α was induced after the burn stimulation. In addition, Hsp90α level was the highest at 12 h post-treatment (C) and somewhat decreased at 48 h (E). Magnification of red boxed areas in (A), (C) and (E) was shown as (B), (D)and (F), respectively.

Mentions: To determine whether a correlation existed between burn injury and Hsp90α expression, we took three approaches to assessing the change on Hsp90α expression after burn injury. Firstly, real-time PCR analysis was used to assess the transcriptional level of Hsp90α during early stages of wound healing. Results indicated that Hsp90α mRNA level in burned skin peaked at 3∼6 h and were 20 times higher than that in unburned mice (Fig. 2A, n = 3 for each time point, p<0.05). Secondly, Hsp90α protein level was increased significantly from 12 h after burn injury and maintained high till 72 h as shown in Fig. 2B. These results provided direct evidences that burn injury could stimulate wound edge area to produce more Hsp90α at both mRNA and protein levels. Previous studies on the function of Hsp90α in wound healing suggested that Hsp90α was more likely to be secreted out to the extracellular space, where it exerted important biological functions [30]. Thirdly, immunohistochemistry was performed to visualize the change on Hsp90α expression after burn treatment (Fig. 3). The burned area showed a high density of Hsp90α immunoreactive staining while the control had less Hsp90α expression. The contrast was most notable at 12 h after burns, with a slight reduction of Hsp90α level at 48 h.


Role for heat shock protein 90α in the proliferation and migration of HaCaT cells and in the deep second-degree burn wound healing in mice.

Zhang Y, Bai X, Wang Y, Li N, Li X, Han F, Su L, Hu D - PLoS ONE (2014)

IHC assay showing the changes of Hsp90α immunostaining in burned mouse skin.(A) Unwound normal skin showed a few positive Hsp90α stainings. (C, E) Epidermal and dermal tissues after burn injury appeared more positive brownish stainings, indicating that Hsp90α was induced after the burn stimulation. In addition, Hsp90α level was the highest at 12 h post-treatment (C) and somewhat decreased at 48 h (E). Magnification of red boxed areas in (A), (C) and (E) was shown as (B), (D)and (F), respectively.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4128658&req=5

pone-0103723-g003: IHC assay showing the changes of Hsp90α immunostaining in burned mouse skin.(A) Unwound normal skin showed a few positive Hsp90α stainings. (C, E) Epidermal and dermal tissues after burn injury appeared more positive brownish stainings, indicating that Hsp90α was induced after the burn stimulation. In addition, Hsp90α level was the highest at 12 h post-treatment (C) and somewhat decreased at 48 h (E). Magnification of red boxed areas in (A), (C) and (E) was shown as (B), (D)and (F), respectively.
Mentions: To determine whether a correlation existed between burn injury and Hsp90α expression, we took three approaches to assessing the change on Hsp90α expression after burn injury. Firstly, real-time PCR analysis was used to assess the transcriptional level of Hsp90α during early stages of wound healing. Results indicated that Hsp90α mRNA level in burned skin peaked at 3∼6 h and were 20 times higher than that in unburned mice (Fig. 2A, n = 3 for each time point, p<0.05). Secondly, Hsp90α protein level was increased significantly from 12 h after burn injury and maintained high till 72 h as shown in Fig. 2B. These results provided direct evidences that burn injury could stimulate wound edge area to produce more Hsp90α at both mRNA and protein levels. Previous studies on the function of Hsp90α in wound healing suggested that Hsp90α was more likely to be secreted out to the extracellular space, where it exerted important biological functions [30]. Thirdly, immunohistochemistry was performed to visualize the change on Hsp90α expression after burn treatment (Fig. 3). The burned area showed a high density of Hsp90α immunoreactive staining while the control had less Hsp90α expression. The contrast was most notable at 12 h after burns, with a slight reduction of Hsp90α level at 48 h.

Bottom Line: The hypoxic wound microenvironment promotes cell migration through a hypoxia--heat shock protein 90 alpha (Hsp90α)--low density lipoprotein receptor-related protein-1 (LRP-1) autocrine loop.Experiments performed with a human keratinocyte cell line--HaCaT also confirmed above results. 17-dimethylaminoethylamino-17demethoxygeldanamycin hydrochloride (17-DMAG), an Hsp90α inhibitor, was used to further evaluate the function of Hsp90α in wound healing.In conclusion, these results provided a rationale for the therapeutic effect of Hsp90α on the burn wound management.

View Article: PubMed Central - PubMed

Affiliation: Department of Burns and Cutaneous Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shannxi, China.

ABSTRACT
Inflammation, proliferation, and tissue remodeling are essential steps for wound healing. The hypoxic wound microenvironment promotes cell migration through a hypoxia--heat shock protein 90 alpha (Hsp90α)--low density lipoprotein receptor-related protein-1 (LRP-1) autocrine loop. To elucidate the role of this autocrine loop on burn wound healing, we investigated the expression profile of Hsp90α at the edge of burn wounds and found a transient increase in both mRNA and protein levels. Experiments performed with a human keratinocyte cell line--HaCaT also confirmed above results. 17-dimethylaminoethylamino-17demethoxygeldanamycin hydrochloride (17-DMAG), an Hsp90α inhibitor, was used to further evaluate the function of Hsp90α in wound healing. Consistently, topical application of Hsp90α in the early stage of deep second-degree burn wounds led to reduced inflammation and increased tissue granulation, with a concomitant reduction in the size of the wound at each time point tested (p<0.05). Consequently, epidermal cells at the wound margin progressed more rapidly causing an expedited healing process. In conclusion, these results provided a rationale for the therapeutic effect of Hsp90α on the burn wound management.

Show MeSH
Related in: MedlinePlus