延寿液延缓臭氧吸入诱导大鼠皮肤衰老的作用机制研究
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摘要
目的:探讨延寿液通过调控炎症和维持胶原延缓臭氧吸入所诱导大鼠皮肤衰老的作用机制。方法:将48只大鼠按照随机数字表法分为对照组、模型组、维生素E组、延寿液低剂量组、延寿液中剂量组和延寿液高剂量组,每组各8只。对照组大鼠每天吸入新鲜空气10 h,模型组和各给药组大鼠每天吸入臭氧(1. 2±0. 2) ppm,10 h造模。检测各组大鼠血浆/肝肾组织总超氧化物歧化酶(T-SOD)和铜锌超氧化物歧化酶(CuZn-SOD)活力、血浆锰超氧化物歧化酶(MnSOD)活力,血清/肝肾组织丙二醛(malondialdeyde,MDA)含量,皮肤羟脯氨酸(Hyp)含量,苦味酸天狼星红染色定量皮肤胶原面积,免疫组织化学染色检测转化生长因子-β(transforming growth factor-β,TGF-β)表达,蛋白斑点测定皮肤基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)表达变化。结果:与对照组比较,模型组大鼠血液T-SOD、MnSOD的活性及Hyp含量下降,MDA含量显著升高(P <0. 01)。与模型组比较,延寿液低剂量组、延寿液中剂量组和延寿液高剂量组T-SOD、Mn-SOD的活性显著升高,CuZn-SOD活性、MDA及Hyp含量显著降低,差异具有统计学意义(P <0. 01)。与对照组比较,模型组肝组织MDA含量升高,肾组织T-SOD、CuZn-SOD活力降低,MDA含量明显增高(P <0. 01);与模型组比较,延寿液低剂量组肝MDA含量降低显著(P <0. 01),延寿液低剂量组、中剂量组肾T-SOD活力显著升高(P <0. 05),延寿液低剂量组肾MDA的含量显著降低(P <0. 05);与对照组比较,模型组TGF-β1含量降低、皮肤MMP-1蛋白表达显著增高,差异有统计学意义(P <0. 01);与模型组比较,维生素E组和延寿液低剂量组中TGF-β1含量升高、皮肤MMP-1的表达降低,差异具有统计学意义(P <0. 05)。结论:延寿液通过上调TGF-β1调控炎症,同时下调MMP-1维持胶原,具有延缓皮肤衰老的作用。
Objective: To explore the mechanism of Yanshou Decoction( YSD) in delaying skin aging in rats by regulating inflammation and maintaining collagen. Methods: Forty-eight rats were randomly divided into the control group,the model group,the vitamin E group,low-dose YSD group,high-dose YSD group and high-dose YSD group,with 8 rats in each group. The rats in the control group were inhaled fresh air for 10 hours every day,and the rats in the model group and the administration groups were inhaled ozone( 1. 2 ± 0. 2) ppm every day for 10 hours. The change of total superoxide dismutase( T-SOD) in plasma/liver and kidney tissues,activity of copper-zinc superoxide dismutase( Cu Zn-SOD),the activity of plasma manganese superoxide dismutase( Mn-SOD),the content of malondialdehyde( MDA) in serum/liver and kidney tissue,the skin hydroxyproline( Hyp) content,quantitative collagen area of skin with Sirius picrate red staining in rats of each group were detected. The expression of transforming growth factor-beta( TGF-β) was detected by immunohistochemical staining and the expression of matrix metalloproteinase-1( MMP-1) in skin was detected by protein spot assay. Results: Compared with the control group,the activity of T-SOD,Mn-SOD and the content of Hyp in the blood of the model group's rats decreased,while the content of MDA increased significantly( P <0. 01). Compared with model group,the activities of T-SOD and Mn-SOD in low dose group,high dose group and high dose group of YSD increased significantly( P < 0. 01),while their activities of Cu Zn-SOD,MDA and Hyp decreased significantly,and all the differences were statistically significant( P < 0. 01). Compared with the control group,MDA content in liver tissue increased,while T-SOD and Cu Zn-SOD activity in kidney tissue decreased and MDA content increased significantly in the model group( P <0. 01). Compared with the model group,the MDA content in liver decreased significantly( P < 0. 01),while the T-SOD activity in kidney increased significantly( P < 0. 05) and the MDA content in kidney decreased significantly( P < 0. 05) in the low-dose group and the medium-dose group of YSD. Compared with the control group,the content of TGF-β1 decreased and the expression of MMP-1 protein increased significantly in the model group,and all the differences were statistically significant( P < 0. 01).Compared with the model group,the content of TGF-β1 increased and the expression of MMP-1 decreased in the vitamin E group and the low-dose YSD group and all the differences were statistically significant( P < 0. 05). Conclusion: YSD has the effect of delaying skin aging by up-regulating TGF-β1 and down-regulating MMP-1 to maintain collagen.
Objective: To explore the mechanism of Yanshou Decoction( YSD) in delaying skin aging in rats by regulating inflammation and maintaining collagen. Methods: Forty-eight rats were randomly divided into the control group,the model group,the vitamin E group,low-dose YSD group,high-dose YSD group and high-dose YSD group,with 8 rats in each group. The rats in the control group were inhaled fresh air for 10 hours every day,and the rats in the model group and the administration groups were inhaled ozone( 1. 2 ± 0. 2) ppm every day for 10 hours. The change of total superoxide dismutase( T-SOD) in plasma/liver and kidney tissues,activity of copper-zinc superoxide dismutase( Cu Zn-SOD),the activity of plasma manganese superoxide dismutase( Mn-SOD),the content of malondialdehyde( MDA) in serum/liver and kidney tissue,the skin hydroxyproline( Hyp) content,quantitative collagen area of skin with Sirius picrate red staining in rats of each group were detected. The expression of transforming growth factor-beta( TGF-β) was detected by immunohistochemical staining and the expression of matrix metalloproteinase-1( MMP-1) in skin was detected by protein spot assay. Results: Compared with the control group,the activity of T-SOD,Mn-SOD and the content of Hyp in the blood of the model group's rats decreased,while the content of MDA increased significantly( P <0. 01). Compared with model group,the activities of T-SOD and Mn-SOD in low dose group,high dose group and high dose group of YSD increased significantly( P < 0. 01),while their activities of Cu Zn-SOD,MDA and Hyp decreased significantly,and all the differences were statistically significant( P < 0. 01). Compared with the control group,MDA content in liver tissue increased,while T-SOD and Cu Zn-SOD activity in kidney tissue decreased and MDA content increased significantly in the model group( P <0. 01). Compared with the model group,the MDA content in liver decreased significantly( P < 0. 01),while the T-SOD activity in kidney increased significantly( P < 0. 05) and the MDA content in kidney decreased significantly( P < 0. 05) in the low-dose group and the medium-dose group of YSD. Compared with the control group,the content of TGF-β1 decreased and the expression of MMP-1 protein increased significantly in the model group,and all the differences were statistically significant( P < 0. 01).Compared with the model group,the content of TGF-β1 increased and the expression of MMP-1 decreased in the vitamin E group and the low-dose YSD group and all the differences were statistically significant( P < 0. 05). Conclusion: YSD has the effect of delaying skin aging by up-regulating TGF-β1 and down-regulating MMP-1 to maintain collagen.
引文
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[5]刘甜甜,张微,彭金娥,等.延寿液对大鼠吸入臭氧诱导皮肤衰老的延缓作用[J].中国实验方剂学杂志,2015,21(7):142-147.
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[16]王昌禄,曹俊武,王玉荣,等.铜锌超氧化物歧化酶(SOD)研究进展:从基因到功能[J].现代生物医学进展,2008,8(5):940-943.
[17]何颖,李冬梅,陶文雁,等.不同类型音乐对老年大鼠学习记忆能力及免疫机能的影响[J].中国免疫学杂志,2014,30(2):182-186.
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[20] CANDELARIO-JALIL E,YANG Y,ROSENBERG G A. Diverse roles of matrix metalloproteinases and tissue inhibitors of metalloproteinases in neuroinflammation and cerebral ischemia[J]. Neuroscience,2009,158(3):983-994.
[21] CUTLER S J,DOECKE J D,GHAZAWI I,et al. Novel STAT binding elements mediate IL-6 regulation of MMP-1 and MMP-3[J]. Sci Rep,2017,7(1):8526.
[22]刘月恒,来吉祥,董银卯,等.基质金属蛋白酶-1与皮肤衰老的研究进展[J].中国老年学杂志,2010,30(20):3037-3039.
[23] HE J W,CAO W W,AZEEM I,et al. Transforming Growth Factor Beta1 being considered a novel biomarker in knee osteoarthritis[J]. Clin Chim Acta,2017,472(6):96-101.
[24] SUN X,LIU W,CHENG G,et al. The influence of connective tissue growth factor on rabit ligament injury repair[J]. Bone Joint Res,2017,6(7):399-404.
[25] YANG Q,XIE R J,YANG T,et al. Transforming growth factorbeta1 and Smad4 signaling pathway down-regulates renal extracellular matrix degradation in diabetic rats[J]. Chin Med Sci J,2007,22(4):243-249.
[26] EL-DOMYATI M,ATTIA S,SALEH F,et al. Intrinsic aging vs.photoaging:A comparative histopathological,immunohistochemical,and ultrastructural study of skin[J]. Exp Dermatol,2002,11(5):398-405.
[2] POLJSAK B,DAHMANE R G,GODIC A. Intrinsic skin aging:the role of oxidative stress[J]. Acta Dermatovenerol Alp Pannonica Adriat,2012,21(2):33-36.
[3]苏明瑛,贾玉杰.皮肤老化的原因及发生机制[J].中国麻风皮肤病杂志,2013,29(8):523-525.
[4] MORITA A,TORII K,MAEDA A,et al. Molecular basis of tobacco smoke-induced premature skin aging[J]. J Investig Dermatol Symp Proc,2009,14(1):53-55.
[5]刘甜甜,张微,彭金娥,等.延寿液对大鼠吸入臭氧诱导皮肤衰老的延缓作用[J].中国实验方剂学杂志,2015,21(7):142-147.
[6]李晓霞.从肝论治女性皮肤衰老探讨[J].中医研究,2014,27(6):8-10.
[7]彭红华.肾与皮肤衰老关系的探析[J].广西中医药,2013,36(3):57-58.
[8]孟姣,吕振宇,孙传鑫,等.枸杞多糖药理作用研究进展[J].时珍国医国药,2018,29(10):2489-2493.
[9]唐冕,许晓芬.药用黄芪皂苷类化学成分及药理作用研究进展[J].中医药导报,2018,24(20):117-122.
[10]姚红旗,侯雅竹,王贤良,等.黄芪心血管药理作用研究进展[J].河南中医,2019,39(2):302-306.
[11]孙凯,蒋伟文.鼠衰老模型的建模方法概述[J].中华老年口腔医学杂志,2017,15(3):181-184.
[12] HARMAN D. Aging:a theory based on free radical and radiation chemistry[J]. J Gerontol,1956,11(3):298-300.
[13]朱润芝,李京敬,谢超,等.过氧化作用与肝脏疾病[J].世界华人消化杂志,2010,18(11):1134-1140.
[14] JOHNSON F,GIULIVI C. Superoxide dismutases and their impact upon human health[J]. Mol Aspects Med,2005,26(4/5):340-352.
[15] RODRGUEZ-MARTNEZ E,MARTNEZ F,ESPINOSA-GARCA M T,et al. Mitochondrial dysfunction in the hippocampus of rats caused by chronic oxidative stress[J]. Neuroscience,2013,252(4):384-395.
[16]王昌禄,曹俊武,王玉荣,等.铜锌超氧化物歧化酶(SOD)研究进展:从基因到功能[J].现代生物医学进展,2008,8(5):940-943.
[17]何颖,李冬梅,陶文雁,等.不同类型音乐对老年大鼠学习记忆能力及免疫机能的影响[J].中国免疫学杂志,2014,30(2):182-186.
[18] VISSE R,NAGASE H. Matrix metalloproteinases and tissue inhibitors of metalloproteinases:structure,function,and bioehemistry[J]. Cire Res,2003,92(8):827-839.
[19] FROST J,RAMSAY M,MIA R,et al. Differential gene expression of MMP-1,TIMP-1 and HGF in clinically involved and uninvolved skin in South Africans with SSc[J]. Rheumatology(Oxford),2012,51(6):1049-1052.
[20] CANDELARIO-JALIL E,YANG Y,ROSENBERG G A. Diverse roles of matrix metalloproteinases and tissue inhibitors of metalloproteinases in neuroinflammation and cerebral ischemia[J]. Neuroscience,2009,158(3):983-994.
[21] CUTLER S J,DOECKE J D,GHAZAWI I,et al. Novel STAT binding elements mediate IL-6 regulation of MMP-1 and MMP-3[J]. Sci Rep,2017,7(1):8526.
[22]刘月恒,来吉祥,董银卯,等.基质金属蛋白酶-1与皮肤衰老的研究进展[J].中国老年学杂志,2010,30(20):3037-3039.
[23] HE J W,CAO W W,AZEEM I,et al. Transforming Growth Factor Beta1 being considered a novel biomarker in knee osteoarthritis[J]. Clin Chim Acta,2017,472(6):96-101.
[24] SUN X,LIU W,CHENG G,et al. The influence of connective tissue growth factor on rabit ligament injury repair[J]. Bone Joint Res,2017,6(7):399-404.
[25] YANG Q,XIE R J,YANG T,et al. Transforming growth factorbeta1 and Smad4 signaling pathway down-regulates renal extracellular matrix degradation in diabetic rats[J]. Chin Med Sci J,2007,22(4):243-249.
[26] EL-DOMYATI M,ATTIA S,SALEH F,et al. Intrinsic aging vs.photoaging:A comparative histopathological,immunohistochemical,and ultrastructural study of skin[J]. Exp Dermatol,2002,11(5):398-405.