Ultraviolet radiation by its wavelength is divided into: UVA, UVB and UVC. Only UVA and UVB manage to penetrate the ozone layer, but due to anthropological activities, all of them are capable of interacting with human...Ultraviolet radiation by its wavelength is divided into: UVA, UVB and UVC. Only UVA and UVB manage to penetrate the ozone layer, but due to anthropological activities, all of them are capable of interacting with humans to a greater or lesser extent, and can generate adverse effects such as cellular stress when interacting with intra-and extracellular biomolecules. The skin is the first organ in contact with UV radiation, and the stress it generates can be analyzed by the expression of a bioindicator of cellular damage such as Hsp70. Therefore, the objective of the project was: to determine the effect of UVA, UVB and UVC radiation on HaCaT epithelial cells, by analyzing the expression of Hsp70. Materials and methods: HaCaT cells were cultured in vitro, which were irradiated with UVA, UVB and UVC light at different doses, to subsequently determine the degree of Hsp70 expression by Immunodetection by PAGE-SDS and Western Blot. Results: Basal expression of Hsp70 was observed in no irradiated HaCaT cells. When HaCaT cells were irradiated with UVA, UVB, UVC, an increase in this Hsp70 protein was observed. With UVA, a higher degree of expression was observed at a time of 30 minutes of irradiation. With UVB the highest expression shifted to a time of 20 minutes. With UVC, overexpression was observed after 10 minutes. Conclusion: UV radiation generates cellular stress on HaCaT cells, evaluated by the stress bioindicator Hsp70. According to the wavelength of UV radiation, those that have a shorter wavelength have a greater potential for cellular damage, such as UVC.展开更多
Background:Amarogentin(AMA)is a secoiridoid glycoside extracted from Swertia and Gentiana roots and exhibits many biological effects such as antioxidative,antiinflammatory,and antitumor activities.Atopic dermatitis(AD...Background:Amarogentin(AMA)is a secoiridoid glycoside extracted from Swertia and Gentiana roots and exhibits many biological effects such as antioxidative,antiinflammatory,and antitumor activities.Atopic dermatitis(AD)is a chronic inflammatory skin disease caused by disorders in the regulation of multiple inflammatory cytokines.No effective cure has been found for AD now.Methods:We constructed the HaCat and splenocyte model and tested the inhibitory effect of AMA on IL-4,IL-6,and IL-13 secretions using enzyme-linked immunosorbent assay(ELISA).The AD mouse model was constructed and treated with AMA,the severity of skin lesions was observed,epidermal tissue was collected,and epidermal thickness and mast cell infiltration were observed using hematoxylin and eosin and toluidine blue staining,respectively.The expression of kallikrein-related peptidase 7(KLK7)and filaggrin(FLG)was detected using immunostaining and Western blot analysis.The mRNA expression of KLK7 and FLG was detected using quantitative polymerase chain reaction(qPCR).Blood immunoglobulin E(IgE)secretion was detected.Results:AMA inhibited IL-6 secreted by tumor necrosis factor(TNF)-α-induced HaCaT cells and reduced IL-4 and IL-13 secreted by phytohemagglutinin(PHA)-induced primary cells in the mice spleen.It was found that the treatment of AMA with 2,4-din itrochlorobenzene-induced AD-like mice could promote the recovery of dermatitis,reduce the score of dermatitis severity and the scratching frequency,treat the skin lesions,reduce the epidermal thickness,decrease the infiltration of mast cells,reduce the IgE level in serum,decrease the expression levels of AD-related cytokines,increase protein and mRNA expression of FLG,and reduce the protein and mRNA expression of KLK7 in the skin tissues of AD-like mice.Conclusion:In conclusion,AMA inhibits inflammatory response at the cellular level,and AMA reduces the validation response of specific dermatitis mice,relieves pruritus,and repairs the damaged skin barrier.展开更多
文摘Ultraviolet radiation by its wavelength is divided into: UVA, UVB and UVC. Only UVA and UVB manage to penetrate the ozone layer, but due to anthropological activities, all of them are capable of interacting with humans to a greater or lesser extent, and can generate adverse effects such as cellular stress when interacting with intra-and extracellular biomolecules. The skin is the first organ in contact with UV radiation, and the stress it generates can be analyzed by the expression of a bioindicator of cellular damage such as Hsp70. Therefore, the objective of the project was: to determine the effect of UVA, UVB and UVC radiation on HaCaT epithelial cells, by analyzing the expression of Hsp70. Materials and methods: HaCaT cells were cultured in vitro, which were irradiated with UVA, UVB and UVC light at different doses, to subsequently determine the degree of Hsp70 expression by Immunodetection by PAGE-SDS and Western Blot. Results: Basal expression of Hsp70 was observed in no irradiated HaCaT cells. When HaCaT cells were irradiated with UVA, UVB, UVC, an increase in this Hsp70 protein was observed. With UVA, a higher degree of expression was observed at a time of 30 minutes of irradiation. With UVB the highest expression shifted to a time of 20 minutes. With UVC, overexpression was observed after 10 minutes. Conclusion: UV radiation generates cellular stress on HaCaT cells, evaluated by the stress bioindicator Hsp70. According to the wavelength of UV radiation, those that have a shorter wavelength have a greater potential for cellular damage, such as UVC.
基金The present study was supported by the National Natural Science Foundation of China(grant numbers:81902067 and 82078189)the Medical Scientific Research Foundation of Guangdong Province(grant number:A2019502)+2 种基金the Shenzhen Science and Technology Innovation Committee(grant numbers:JCY20180305124849781 and 20200812211704001)the SZU Top Ranking Project(grant number:86000000210)the Hospital Project of Huazhong University of Science and Technology Union Shenzhen Hospital(grant number:2021042).
文摘Background:Amarogentin(AMA)is a secoiridoid glycoside extracted from Swertia and Gentiana roots and exhibits many biological effects such as antioxidative,antiinflammatory,and antitumor activities.Atopic dermatitis(AD)is a chronic inflammatory skin disease caused by disorders in the regulation of multiple inflammatory cytokines.No effective cure has been found for AD now.Methods:We constructed the HaCat and splenocyte model and tested the inhibitory effect of AMA on IL-4,IL-6,and IL-13 secretions using enzyme-linked immunosorbent assay(ELISA).The AD mouse model was constructed and treated with AMA,the severity of skin lesions was observed,epidermal tissue was collected,and epidermal thickness and mast cell infiltration were observed using hematoxylin and eosin and toluidine blue staining,respectively.The expression of kallikrein-related peptidase 7(KLK7)and filaggrin(FLG)was detected using immunostaining and Western blot analysis.The mRNA expression of KLK7 and FLG was detected using quantitative polymerase chain reaction(qPCR).Blood immunoglobulin E(IgE)secretion was detected.Results:AMA inhibited IL-6 secreted by tumor necrosis factor(TNF)-α-induced HaCaT cells and reduced IL-4 and IL-13 secreted by phytohemagglutinin(PHA)-induced primary cells in the mice spleen.It was found that the treatment of AMA with 2,4-din itrochlorobenzene-induced AD-like mice could promote the recovery of dermatitis,reduce the score of dermatitis severity and the scratching frequency,treat the skin lesions,reduce the epidermal thickness,decrease the infiltration of mast cells,reduce the IgE level in serum,decrease the expression levels of AD-related cytokines,increase protein and mRNA expression of FLG,and reduce the protein and mRNA expression of KLK7 in the skin tissues of AD-like mice.Conclusion:In conclusion,AMA inhibits inflammatory response at the cellular level,and AMA reduces the validation response of specific dermatitis mice,relieves pruritus,and repairs the damaged skin barrier.
