脂肪来源干细胞调控皮肤光老化的细胞因子分析
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摘要
目的通过对蛋白芯片研究以及文献复习探索脂肪来源干细胞(adipose-derived stem cells,ADSCs)分泌的多种细胞因子中,参与调控缓解光老化的关键性细胞因子。方法体外培养小鼠ADSCs及皮肤成纤维细胞,提取胞浆蛋白,ELISA测定胞浆蛋白中肝细胞生长因子(hepatocyte growth factor,HGF)的表达量;利用蛋白芯片技术筛选出表达差异的细胞因子。文献复习将筛选出的细胞因子作用于小鼠皮肤光老化模型,如能缓解光老化表型,则为ADSCs发挥抗老化作用的关键细胞因子。结果 GSM-CAA-4000蛋白芯片能够检测的200种细胞因子/蛋白中,ADSCs及成纤维细胞裂解液中细胞因子/蛋白水平存在显著差异的细胞因子/蛋白有53种。这53种细胞因子/蛋白与对照组成纤维细胞相比,ADSCs分泌占优势的有38种。其中HGF以及血小板衍生生长因子(platelet-derived growth factor,PDFG)对光老化模型具有拮抗作用。结论筛选出的差异性细胞因子以HGF及PDGF为代表,在拮抗光老化过程中作为关键因子发挥重要的作用。
Objective Through the research of protein antibody array and review of related literature, to explore the key cytokines involved in the regulation of photoaging which secreted by adipose-derived stem cells(ADSCs). Methods ADSCs and skin fibroblasts(FBs) of mice were cultured in vitro, then cytoplasm proteins were extracted. The expression of hepatocyte growth factor(HGF) in cytoplasm protein was detected by ELISA. The differentially expressed cytokine factors were screened through protein antibody array technology. The mice skin photoaging model was applied to cytokines screened through literature review. If these cytokines can alleviate the photoaging phenotype, it will be a key cytokine for anti-aging effects of ADSCs. Results Among the 200 kinds of cytokines/proteins detected by GSM-CAA-4000 protein antibody array, there were 53 kinds of cytokines/proteins with significant differences in cytokine/protein levels in ADSCs and FBs lysates. The 53 cytokines/proteins were dominant in the secretion of ADSCs compared to the control fibroblasts. Among them, HGF and platelet-derived growth factor(PDGF) have antagonistic effects on the photoaging model. Conclusion The screened differential cytokines, such as HGF and PDGF, play an important role in the process of antagonizing photoaging.
Objective Through the research of protein antibody array and review of related literature, to explore the key cytokines involved in the regulation of photoaging which secreted by adipose-derived stem cells(ADSCs). Methods ADSCs and skin fibroblasts(FBs) of mice were cultured in vitro, then cytoplasm proteins were extracted. The expression of hepatocyte growth factor(HGF) in cytoplasm protein was detected by ELISA. The differentially expressed cytokine factors were screened through protein antibody array technology. The mice skin photoaging model was applied to cytokines screened through literature review. If these cytokines can alleviate the photoaging phenotype, it will be a key cytokine for anti-aging effects of ADSCs. Results Among the 200 kinds of cytokines/proteins detected by GSM-CAA-4000 protein antibody array, there were 53 kinds of cytokines/proteins with significant differences in cytokine/protein levels in ADSCs and FBs lysates. The 53 cytokines/proteins were dominant in the secretion of ADSCs compared to the control fibroblasts. Among them, HGF and platelet-derived growth factor(PDGF) have antagonistic effects on the photoaging model. Conclusion The screened differential cytokines, such as HGF and PDGF, play an important role in the process of antagonizing photoaging.
引文
[1]王婷,郭澍,陈鹏,等.脂肪干细胞条件培养基对不同衰老程度成纤维细胞中波紫外线损伤的修复效应[J].中华医学美学美容杂志, 2018,24(2):79-83.
[2] LEE S H, JIN S Y, SONG J S, et al. Paracrine effects of adiposederived stem cells on keratinocytes and dermal fibro-blasts[J]. Ann Dermatol, 2012,24(2):136-143.
[3] YANG J A, CHUNG H M, WON C H, et al. Potential application of adipose-derived stem cells and their secretory factors to skin:discussion from both clinical and industrial viewpoints[J]. Expert Opin Biol Ther, 2010,10(4):495-503.
[4] HSIAO S T, ASGARI A, LOKMIC Z, et al. Comparative analysis of paracrine factor expression in human adult mesenchymal stem cells derived from bone marrow, adipose, and dermal tissue[J]. Stem Cells Dev, 2012,21(12):2189-2203.
[5] MOON K M, PARK Y H, LEE J S, et al. The effect of secretory factors of adipose-derived stem cells on human keratinocytes[J].Int J Mol Sci, 2012,13(1):1239-1257.
[6] CHEN Y T, SUN C K, LIN Y C, et al. Adipose-derived mesenchymal stem cell protects kidneys against ischemia-reperfusion injury through suppressing oxidative stress and inflammatory reaction[J].J Transl Med, 2011,9:51.
[7] KIM J H, JUNG M, KIM H S, et al. Adipose-derived stem cells as a new therapeutic modality for ageing skin[J]. Exp Dermatol, 2011,20(5):383-387.
[8] KIM W S, PARK B S, PARK S H, et al. Antiwrinkle effect of adipose-derived stem cell:activation of dermal fibroblast by secretory factors[J]. J Dermatol Sci, 2009,53(2):96-102.
[9] KIM W S, PARK B S, SUNG J H, et al. Wound healing effect of adipose-derived stem cells:a critical role of secretory factors on human dermal fibroblasts[J]. J Dermatol Sci, 2007,48(1):15-24.
[10] KIM W S, PARK B S, KIM H K, et al. Evidence supporting antioxidant action of adipose-derived stem cells:protection of human dermal fibroblasts from oxidative stress[J]. J Dermatol Sci,2008,49(2):133-142.
[11] KIM W S, PARK B S, SUNG J H. Protective role of adipose-derived stem cells and their soluble factors in photoaging[J]. Arch Dermatol Res, 2009,301(5):329-336.
[12]曾继平,毕波,陈亮,等.脂肪来源干细胞对光老化成纤维细胞的治疗作用[J].中国美容整形外科杂志, 2013,24(10):581-586.
[13] JEONG J H, FAN Y, YOU G Y, et al. Improvement of photoaged skin wrinkles with cultured human fibroblasts and adipose-derived stem cells:a comparative study[J]. J Plast Reconstr Aesthet Surg,2015,68(3):372-381.
[14]杨清建,毕波,陈亮,等.小鼠脂肪来源间充质干细胞细胞因子蛋白质组学分析[J].中国美容整形外科杂志, 2016,27(2):79-81.
[15] ZENG J P, BI B, CHEN L, et al. Repeated exposure of mouse dermal fibroblasts at a sub-cytotoxic dose of UVB leads to premature senescence:a robust model of cellular photoaging[J]. J Dermatol Sci, 2014,73(1):49-56.
[16]杨清建,毕波,贾传龙,等.肝细胞生长因子对紫外线诱导皮肤成纤维细胞产生活性氧的影响[J].老年医学与保健, 2017,23(2):118-121.
[17]任润健,方磊,贾传龙,等. PDGF-BB对UVB诱导的光老化成纤维细胞中ROS的影响[J].中国美容医学, 2018,27(5):109-112.
[18] YAMABA H, HABA M, KUNITA M, et al. Morphological change of skin fibroblasts induced by UV Irradiation is involved in photoaging[J]. Exp Dermatol, 2016,25 Suppl 3:45-51.
[19] SANADA F, TANIYAMA Y, AZUMA J, et al. Hepatocyte growth factor, but not vascular endothelial growth factor, attenuates angiotensin II-induced endothelial progenitor cell senescence[J].Hypertension, 2009,53(1):77-82.
[20] HUI L, HONG Y, JINGJING Z, et al. HGF suppresses high glucose-mediated oxidative stress in mesangial cells by activation of PKG and inhibition of PKA[J]. Free Radic Biol Med, 2010,49(3):467-473.
[21] KITTA K, DAY R M, IKEDA T, et al. Hepatocyte growth factor protects cardiac myocytes against oxidative stress-induced apoptosis[J]. Free Radic Biol Med, 2001,31(7):902-910.
[22] FLUSBERG D A, SORGER P K. Surviving apoptosis:life-death signaling in single cells[J]. Trends Cell Biol, 2015,25(8):446-458.
[23]杨清建,毕波,朱宁文,等.肝细胞生长因子对光老化成纤维细胞凋亡及周期的影响[J].中国美容整形外科杂志, 2016,28(12):740-743.
[24] Fredriksson L, Li H, Eriksson U. The PDGF family:four gene products form five dimeric isoforms[J]. Cytokine Growth Factor Rev,2004,15(4):197-204.
[25] Salabei J K, Cummins T D, Singh M, et al. PDGF-mediated autophagy regulates vascular smooth muscle cell phenotype and resistance to oxidative stress[J]. Biochem J, 2013,451(3):375-388.
[26] Zheng L, Ishii Y, Tokunaga A, et al. Neuroprotective effects of PDGF against oxidative stress and the signaling pathway involved[J].J Neurosci Res, 2010,88(6):1273-1284.
[27]贾传龙,杨清建,毕波,等.富血小板血浆促进光老化成纤维细胞分泌功能的研究[J].老年医学与保健, 2018,24(1):62-66.
[28]贾传龙,陈亮,杨清建,等.富血小板血浆对光老化皮肤成纤维细胞增殖的影响[J].中国美容整形外科杂志, 2016,28(4):234-237.
[29]贾传龙,杨清建,毕波,等.富血小板血浆对中波紫外线照射成纤维细胞活性氧的影响[J].中华医学美学美容杂志, 2018,24(1):54-57.
[30] MARTIN G, RICUCCI D, GIBBS J L, et al. Histological findings of revascularized/revitalized immature permanent molar with apical periodontitis using platelet-rich plasma[J]. J Endod, 2013,39(1):138-144.
[2] LEE S H, JIN S Y, SONG J S, et al. Paracrine effects of adiposederived stem cells on keratinocytes and dermal fibro-blasts[J]. Ann Dermatol, 2012,24(2):136-143.
[3] YANG J A, CHUNG H M, WON C H, et al. Potential application of adipose-derived stem cells and their secretory factors to skin:discussion from both clinical and industrial viewpoints[J]. Expert Opin Biol Ther, 2010,10(4):495-503.
[4] HSIAO S T, ASGARI A, LOKMIC Z, et al. Comparative analysis of paracrine factor expression in human adult mesenchymal stem cells derived from bone marrow, adipose, and dermal tissue[J]. Stem Cells Dev, 2012,21(12):2189-2203.
[5] MOON K M, PARK Y H, LEE J S, et al. The effect of secretory factors of adipose-derived stem cells on human keratinocytes[J].Int J Mol Sci, 2012,13(1):1239-1257.
[6] CHEN Y T, SUN C K, LIN Y C, et al. Adipose-derived mesenchymal stem cell protects kidneys against ischemia-reperfusion injury through suppressing oxidative stress and inflammatory reaction[J].J Transl Med, 2011,9:51.
[7] KIM J H, JUNG M, KIM H S, et al. Adipose-derived stem cells as a new therapeutic modality for ageing skin[J]. Exp Dermatol, 2011,20(5):383-387.
[8] KIM W S, PARK B S, PARK S H, et al. Antiwrinkle effect of adipose-derived stem cell:activation of dermal fibroblast by secretory factors[J]. J Dermatol Sci, 2009,53(2):96-102.
[9] KIM W S, PARK B S, SUNG J H, et al. Wound healing effect of adipose-derived stem cells:a critical role of secretory factors on human dermal fibroblasts[J]. J Dermatol Sci, 2007,48(1):15-24.
[10] KIM W S, PARK B S, KIM H K, et al. Evidence supporting antioxidant action of adipose-derived stem cells:protection of human dermal fibroblasts from oxidative stress[J]. J Dermatol Sci,2008,49(2):133-142.
[11] KIM W S, PARK B S, SUNG J H. Protective role of adipose-derived stem cells and their soluble factors in photoaging[J]. Arch Dermatol Res, 2009,301(5):329-336.
[12]曾继平,毕波,陈亮,等.脂肪来源干细胞对光老化成纤维细胞的治疗作用[J].中国美容整形外科杂志, 2013,24(10):581-586.
[13] JEONG J H, FAN Y, YOU G Y, et al. Improvement of photoaged skin wrinkles with cultured human fibroblasts and adipose-derived stem cells:a comparative study[J]. J Plast Reconstr Aesthet Surg,2015,68(3):372-381.
[14]杨清建,毕波,陈亮,等.小鼠脂肪来源间充质干细胞细胞因子蛋白质组学分析[J].中国美容整形外科杂志, 2016,27(2):79-81.
[15] ZENG J P, BI B, CHEN L, et al. Repeated exposure of mouse dermal fibroblasts at a sub-cytotoxic dose of UVB leads to premature senescence:a robust model of cellular photoaging[J]. J Dermatol Sci, 2014,73(1):49-56.
[16]杨清建,毕波,贾传龙,等.肝细胞生长因子对紫外线诱导皮肤成纤维细胞产生活性氧的影响[J].老年医学与保健, 2017,23(2):118-121.
[17]任润健,方磊,贾传龙,等. PDGF-BB对UVB诱导的光老化成纤维细胞中ROS的影响[J].中国美容医学, 2018,27(5):109-112.
[18] YAMABA H, HABA M, KUNITA M, et al. Morphological change of skin fibroblasts induced by UV Irradiation is involved in photoaging[J]. Exp Dermatol, 2016,25 Suppl 3:45-51.
[19] SANADA F, TANIYAMA Y, AZUMA J, et al. Hepatocyte growth factor, but not vascular endothelial growth factor, attenuates angiotensin II-induced endothelial progenitor cell senescence[J].Hypertension, 2009,53(1):77-82.
[20] HUI L, HONG Y, JINGJING Z, et al. HGF suppresses high glucose-mediated oxidative stress in mesangial cells by activation of PKG and inhibition of PKA[J]. Free Radic Biol Med, 2010,49(3):467-473.
[21] KITTA K, DAY R M, IKEDA T, et al. Hepatocyte growth factor protects cardiac myocytes against oxidative stress-induced apoptosis[J]. Free Radic Biol Med, 2001,31(7):902-910.
[22] FLUSBERG D A, SORGER P K. Surviving apoptosis:life-death signaling in single cells[J]. Trends Cell Biol, 2015,25(8):446-458.
[23]杨清建,毕波,朱宁文,等.肝细胞生长因子对光老化成纤维细胞凋亡及周期的影响[J].中国美容整形外科杂志, 2016,28(12):740-743.
[24] Fredriksson L, Li H, Eriksson U. The PDGF family:four gene products form five dimeric isoforms[J]. Cytokine Growth Factor Rev,2004,15(4):197-204.
[25] Salabei J K, Cummins T D, Singh M, et al. PDGF-mediated autophagy regulates vascular smooth muscle cell phenotype and resistance to oxidative stress[J]. Biochem J, 2013,451(3):375-388.
[26] Zheng L, Ishii Y, Tokunaga A, et al. Neuroprotective effects of PDGF against oxidative stress and the signaling pathway involved[J].J Neurosci Res, 2010,88(6):1273-1284.
[27]贾传龙,杨清建,毕波,等.富血小板血浆促进光老化成纤维细胞分泌功能的研究[J].老年医学与保健, 2018,24(1):62-66.
[28]贾传龙,陈亮,杨清建,等.富血小板血浆对光老化皮肤成纤维细胞增殖的影响[J].中国美容整形外科杂志, 2016,28(4):234-237.
[29]贾传龙,杨清建,毕波,等.富血小板血浆对中波紫外线照射成纤维细胞活性氧的影响[J].中华医学美学美容杂志, 2018,24(1):54-57.
[30] MARTIN G, RICUCCI D, GIBBS J L, et al. Histological findings of revascularized/revitalized immature permanent molar with apical periodontitis using platelet-rich plasma[J]. J Endod, 2013,39(1):138-144.
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