脂肪干细胞对大鼠急性放射性皮肤损伤的干预作用
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摘要
目的:研究脂肪干细胞(adipose derived stem cells,ADSCs)在SD大鼠放射性皮肤损伤中的干预作用。方法:采用剂量为90 Gy的192Ir放射源,对SD大鼠左侧大腿内侧皮肤进行单次近距离照射,建立放射性皮肤损伤动物模型,并随机将其分为实验组和对照组(n=9)。照射后24 h,实验组大鼠照射处注射60μL(5×105个)脂肪干细胞,对照组注射等体积的PBS。每隔3～4 d观察大鼠照射处皮肤损伤及愈合情况。照射后第28天,处死所有SD大鼠,取照射处皮肤组织,同时取未照射的正常组(n=9)相应部位的皮肤组织切片,采用免疫组织化学法比较3组皮肤组织中α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达情况;HE染色观察皮肤组织病理学表现。结果:SD大鼠在受照射后出现III或IV度放射性皮肤损伤;实验组的创面评级明显低于对照组,愈合时间明显短于对照组;正常组皮肤组织中不表达α-SMA,实验组较对照组皮肤组织中的α-SMA表达增强;实验组表皮厚度低于对照组,而血管密度明显增加(均P<0.05)。结论:192Ir放射剂量为90 Gy时可以导致大鼠皮肤损伤,而ADSCs可以减轻放射性皮肤损伤,并通过增加微血管新生促进溃疡愈合。
Objective: To investigate the effect of adipose-derived stem cells(ADSCs) on radiation-induced skin injury in SD rats.Methods: Radioactive particles 192 Ir were used to irradiate the left medial th igh skin of SD rats, and the irradiation dose was at 90 Gy. Then, the rats were randomly allocated into a control group and a treatment group(each n=9). After the irradiation, the control group was injected with 60 μL PBS and the treatment group was injected with 60 μL ADSCs in irradiated skin. The progress of skin damage and healing was observed and photographed every day. Twenty-eighth days after the irradiation, the irradiated skin tissue was taken from the left th igh, and then fixed with formaldehyde fixative solution. At the same time, the skin tissue of the corresponding part of the normal group(n=9) th at was not irradiated was also taken. After sampling, embedding and slicing, immunohistochemical staining was used to compare the levels of α-smooth muscle actin(α-SMA), and HE staining was used to compare path ological features of the skin.Results: Radioactive particle 192 Ir caused the development of III or IV radioactive skin damage. The score of the treatment group was significantly lower th an th at of the control group. The wounds of the treatment group were basically healed at 28 days, while the ulcer of the control group was unhealed. So, the healing time was shorter in the treatment group. The expression of α-SMA in the skin of the two groups was increased after the radiotherapy. By analyzing the path ological microstructure image, we found th at the th ickness of epidermis in the control group was greater th an th at in the treatment group, while the vascular density in the treatment group was greater th an th at in the control group(all P<0.05).Conclusion: Radioactive particles 192 Ir can cause skin damage, while the adipose-derived stem cells might alleviate radiation-induced skin injury and promote ulcer healing by promoting angiogenesis.
Objective: To investigate the effect of adipose-derived stem cells(ADSCs) on radiation-induced skin injury in SD rats.Methods: Radioactive particles 192 Ir were used to irradiate the left medial th igh skin of SD rats, and the irradiation dose was at 90 Gy. Then, the rats were randomly allocated into a control group and a treatment group(each n=9). After the irradiation, the control group was injected with 60 μL PBS and the treatment group was injected with 60 μL ADSCs in irradiated skin. The progress of skin damage and healing was observed and photographed every day. Twenty-eighth days after the irradiation, the irradiated skin tissue was taken from the left th igh, and then fixed with formaldehyde fixative solution. At the same time, the skin tissue of the corresponding part of the normal group(n=9) th at was not irradiated was also taken. After sampling, embedding and slicing, immunohistochemical staining was used to compare the levels of α-smooth muscle actin(α-SMA), and HE staining was used to compare path ological features of the skin.Results: Radioactive particle 192 Ir caused the development of III or IV radioactive skin damage. The score of the treatment group was significantly lower th an th at of the control group. The wounds of the treatment group were basically healed at 28 days, while the ulcer of the control group was unhealed. So, the healing time was shorter in the treatment group. The expression of α-SMA in the skin of the two groups was increased after the radiotherapy. By analyzing the path ological microstructure image, we found th at the th ickness of epidermis in the control group was greater th an th at in the treatment group, while the vascular density in the treatment group was greater th an th at in the control group(all P<0.05).Conclusion: Radioactive particles 192 Ir can cause skin damage, while the adipose-derived stem cells might alleviate radiation-induced skin injury and promote ulcer healing by promoting angiogenesis.
引文
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[14]Citrin D, Cotrim AP, Hyodo F, et al. Radioprotectors and mitigatorsof radiation-induced normal tissue injury[J]. Oncologist, 2010,15(4):360-371.
[15]李佳,邢志伟,于程程,等.放射性皮肤损伤研究进展[J].中华临床医师杂志(电子版), 2013, 7(6):2650-2652.LI Jia, XING Zhiwei, YU Chengcheng, et al. Progress in research onradiation skin damage[J]. Chinese Journal of Clinicians. ElectronicEdition, 2013, 7(6):2650-2652.
[16]Dormand EL, Banwell PE, Goodacre TE. Radiotherapy and woundhealing[J]. Int Wound J, 2005, 2(2):112-127.
[17]YarnoldJ,BrotonsMC.Path ogeneticmechanismsinradiationfibrosis[J]. Radiother Oncol, 2010, 97(1):149-161.
[18]Akita S, Akino K, Hirano A, et al. Noncultured autologous adipose-derived stem cells therapy for chronic radiation injury[J]. Stem CellsInt, 2010, 2010:532704.
[19]Borgquist O, Ingemansson R, Malmsjo M. Individualizing the useof negative pressure wound therapy for optimal wound healing:afocused review of the literature[J]. Ostomy Wound Manage, 2011,57(4):44-54.
[20]Zhang Q, Fong CC, Yu WK, et al. Herbal formula Astragali Radixand Rehmanniae Radix exerted wound healing effecton human skinfibroblast cell line Hs27 via the activation of transformation growth factor(TGF-beta)path wayandpromotingextracellularmatrix(ECM)deposition[J]. Phytomedicine, 2012, 20(1):9-16.
[21]屈纪富,程天民,郝利.参与皮肤伤口愈合的细胞及其作用的研究进展[J].现代康复, 2001, 5(24):74-75.QUJifu,CHENGTianmin,HAOLi.Advancesinresearchoncells involved in wound skin healing and their effects[J]. ModernRehabilitation, 2001, 5(24):74-75.
[22]Moulin V, Castilloux G, Jean A, et al. In vitro models to study woundhealing fibroblasts[J]. Burns, 1996, 22(5):359-362.
[23]Moulin V, Castilloux G, Auger FA, et al. Modulated response tocytokines of human wound healing myofibroblasts compared todermal fibroblasts[J]. Exp Cell Res, 1998, 238(1):283-293.
[24]曹卫红,杨志祥,谷庆阳,等.急性放射性皮肤溃疡伤口收缩迟缓的机制[J].中国临床康复, 2002, 6(12):1754-1755.CAOWeihong,YANGZhix iang,GUQing yang,etal.Themechanism of wound contraction retardation of acute radiation-induced skin ulcers[J]. Chinese Journal of Clinical Rehabilitation,2002, 6(12):1754-1755.
[25]毛任翔,祁少海,舒斌,等.骨髓间充质干细胞-透明质酸复合物对放射性溃疡创面愈合的影响[J].中华损伤与修复杂志(电子版), 2009, 4(5):517-525.MAO Renxiang, QI Shaohai, SHU Bin, et al. Effect of mesenchymalstemcells-hyaluronicacidcompositiononthefull-th icknesscutaneous wounds healing of radiation induced ulcer[J]. ChineseJournal of Injury Repair and Wound Healing. Electronic Edition,2009, 4(5):517-525.
[2]邹艳花,廖先珍,许可葵,等. 2009—2012年湖南省肿瘤登记地区鼻咽癌的发病与死亡分析[J].实用预防医学, 2016, 23(12):1415-1418.ZOU Yanhua, LIAO Xianzhen, XU Kekui, et al. Analysis of incidenceand mortality of nasopharyngeal carcinoma in Hunan cancer registryareain2009—2012[J].PracticalPreventionMedicine,2016,23(12):1415-1418.
[3]陈三妹,焦迎春,唐四元,等.鼻咽癌放射治疗患者颈部皮肤护理干预效果比较[J].中华护理杂志, 2013, 48(6):542-545.CHENSanmei,JIAOYingchun,TANGSiyuan,etal.Effectsofdifferent interventions in the prevention and treatment of acuteradiation-inducedskinreactionsinneckamongpatientswith nasopharyngeal cancer[J]. Chinese Journal of Nursing, 2013, 48(6):542-545.
[4]Shu k laP N,Ga i rolaM,Mohant iB K,eta l.Pro phy lac t i cbeclometh asone spray to the skin during postoperative radiotherapyof carcinoma breast:a prospective randomized study[J]. Indian JCancer, 2006, 43(4):180-184.
[5]殷剑明,徐伯平,冯惠霞,等.单味黄芩防治急性放射性皮炎的临床观察[J].中国中西医结合杂志, 2001, 21(4):304.YIN Jianming, XU Boping, FENG Huixia, et al. Clinical observationof single-flavored astragalus against acute radiation dermatitis[J].Chinese Journal of Integrative Medicine, 2001, 21(4):180-184.
[6]Borg M, Wilkinson D, Humeniuk V, et al. Successful treatment ofradiation induced breast ulcer with hyperbaric oxygen[J]. Breast,2001, 10(4):336-341.
[7]Zuk PA, Zhu M, Ashjian P, et al. Human adipose tissue is a source ofmultipotent stem cells[J]. Mol Biol Cell, 2002, 13(12):4279-4295.
[8]Kim WS, Park BS, Sung JH. The wound-healing and antioxidanteffects of adipose-derived stem cells[J]. Expert Opin Biol Ther, 2009,9(7):879-887.
[9]Maharlooei MK, Bagheri M, Solhjou Z, et al. Adipose tissue derivedmesenchymal stem cell(AD-MSC)promotes skin wound healing indiabetic rats[J]. Diabetes Res Clin Pract, 2011, 93(2):228-234.
[10]Teng M, Huang Y, Zhang H. Application of stems cells in woundhealing—an update[J]. Wound Repair Regen, 2014, 22(2):151-160.
[11]You HJ, Han SK. Cell therapy for wound healing[J]. J Korean MedSci, 2014, 29(3):311-319.
[12]杨超,邢新,徐达圆,等.脂肪干细胞-透明质酸复合物促进放射性复合损伤创面愈合的初步研究[J].中国修复重建外科杂志, 2011, 25(12):1499-1503.YANG Chao, XING Xin, XU Dayuan, et al. Adipose-derived stemcells hyaluronic acid radiation combined injury wound healing rat[J].Chinese Journal of Reconstructive Surgery, 2011, 25(12):1499-1503.
[13]Douglas BG, Fowler JF. The effect of multiple small doses of X rayson skin reactions in the mouse and a basic interpretation. 1976[J].Radiat Res, 2012, 178(2):V125-V138.
[14]Citrin D, Cotrim AP, Hyodo F, et al. Radioprotectors and mitigatorsof radiation-induced normal tissue injury[J]. Oncologist, 2010,15(4):360-371.
[15]李佳,邢志伟,于程程,等.放射性皮肤损伤研究进展[J].中华临床医师杂志(电子版), 2013, 7(6):2650-2652.LI Jia, XING Zhiwei, YU Chengcheng, et al. Progress in research onradiation skin damage[J]. Chinese Journal of Clinicians. ElectronicEdition, 2013, 7(6):2650-2652.
[16]Dormand EL, Banwell PE, Goodacre TE. Radiotherapy and woundhealing[J]. Int Wound J, 2005, 2(2):112-127.
[17]YarnoldJ,BrotonsMC.Path ogeneticmechanismsinradiationfibrosis[J]. Radiother Oncol, 2010, 97(1):149-161.
[18]Akita S, Akino K, Hirano A, et al. Noncultured autologous adipose-derived stem cells therapy for chronic radiation injury[J]. Stem CellsInt, 2010, 2010:532704.
[19]Borgquist O, Ingemansson R, Malmsjo M. Individualizing the useof negative pressure wound therapy for optimal wound healing:afocused review of the literature[J]. Ostomy Wound Manage, 2011,57(4):44-54.
[20]Zhang Q, Fong CC, Yu WK, et al. Herbal formula Astragali Radixand Rehmanniae Radix exerted wound healing effecton human skinfibroblast cell line Hs27 via the activation of transformation growth factor(TGF-beta)path wayandpromotingextracellularmatrix(ECM)deposition[J]. Phytomedicine, 2012, 20(1):9-16.
[21]屈纪富,程天民,郝利.参与皮肤伤口愈合的细胞及其作用的研究进展[J].现代康复, 2001, 5(24):74-75.QUJifu,CHENGTianmin,HAOLi.Advancesinresearchoncells involved in wound skin healing and their effects[J]. ModernRehabilitation, 2001, 5(24):74-75.
[22]Moulin V, Castilloux G, Jean A, et al. In vitro models to study woundhealing fibroblasts[J]. Burns, 1996, 22(5):359-362.
[23]Moulin V, Castilloux G, Auger FA, et al. Modulated response tocytokines of human wound healing myofibroblasts compared todermal fibroblasts[J]. Exp Cell Res, 1998, 238(1):283-293.
[24]曹卫红,杨志祥,谷庆阳,等.急性放射性皮肤溃疡伤口收缩迟缓的机制[J].中国临床康复, 2002, 6(12):1754-1755.CAOWeihong,YANGZhix iang,GUQing yang,etal.Themechanism of wound contraction retardation of acute radiation-induced skin ulcers[J]. Chinese Journal of Clinical Rehabilitation,2002, 6(12):1754-1755.
[25]毛任翔,祁少海,舒斌,等.骨髓间充质干细胞-透明质酸复合物对放射性溃疡创面愈合的影响[J].中华损伤与修复杂志(电子版), 2009, 4(5):517-525.MAO Renxiang, QI Shaohai, SHU Bin, et al. Effect of mesenchymalstemcells-hyaluronicacidcompositiononthefull-th icknesscutaneous wounds healing of radiation induced ulcer[J]. ChineseJournal of Injury Repair and Wound Healing. Electronic Edition,2009, 4(5):517-525.