活血解毒方对银屑病小鼠血管增殖以及炎症因子的干预作用
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摘要
目的观察活血解毒方对咪喹莫特诱导的银屑病样小鼠模型皮损的干预作用,明确解毒类中药在银屑病治疗中的重要性。方法 BALB/c雄性小鼠55只,随机分为正常对照组、模型组、活血解毒方组、活血方组和甲氨蝶呤组,每组11只,采用外涂咪喹莫特诱导皮肤银屑病样模型。甲氨蝶呤(1 mg/kg)、活血解毒方组(35.1 g/kg)及活血方组(26.9 g/kg)分别予以药物灌胃,正常对照组和模型组予以等量纯净水灌胃。于第7天观察小鼠皮损变化情况后进行处死,取皮肤组织进行检测。HE染色光镜下观察皮损组织形态学变化、表皮层厚度;免疫组织化学法检测皮损中增殖细胞核抗原(PCNA)和T淋巴细胞表面标志CD3的表达以及微血管阳性标记物CD31的表达并检测微血管密度(MVD);采用实时PCR技术检测皮损白细胞介素-17(IL-17)和白细胞介素-23(IL-23)mRNA的表达。结果 HE染色结果显示活血解毒方组皮损表皮增生较模型组少,角化不全的细胞明显减少;表皮厚度降低,P<0.001;活血解毒方组PCNA、CD3阳性表达远低于模型组,P<0.001;活血解毒方MVD明显低于模型组,P<0.05;活血解毒方组IL-17、IL-23的mRNA表达水平均低于模型组,P<0.05。结论活血解毒方的活血润肤作用是通过抑制血管增殖,而清热解毒作用则通过调节IL-23/IL-17轴减少Th17相关因子的表达、表皮层角质细胞的增生、真皮层免疫细胞浸润以改善咪喹莫特诱导的小鼠银屑病样皮损改变。
Objective To observe the interventional effect of Huoxue Jiedu Fang(Blood-activating Toxin-removing Formula) on skin lesions in psoriasiform mouse model induced by imiquimod, and clarify the importance of detoxification Chinese medicinals in treatment of psoriasis. Methods Male BALB/c mice(n =55) were randomly divided into normal control group, model group, Huoxue Jiedu Fang group(HJF group), Huoxue Fang(Blood-activating Formula) group(HF group) and methotrexate group(MTX group, each n =11). The psoriasiform mouse model was induced by external coating imiquimod. MTX group was orally given MTX(1 mg/kg), HJF group, HJF(35.1 g/kg), HF group, HF(26.9 g/kg), while normal control group and model group, equivalent pure water. All mice were executed on the 7~(th) d for collecting skin tissue samples. The scores of psoriasis area and severity index(PASI) were observed. The histomorphological changes and epidermal layer thickness of skin lesion tissue were observed by using light microscope after HE staining. The expressions of proliferating cell nuclear antigen(PCNA), marker on T-cell(CD3) and positive microvessel marker(CD31), and microvessel density(MVD) were detected by using immunohistochemistry technique. The mRNA expressions of interleukin-17(IL-17) and interleukin-23(IL-23) were detected by using real-time PCR. Results The results of HE staining showed that epidermal proliferation, para-keratosis cells and epidermal thickness decreased in HJF group compared with model group(P<0.001). The positive expressions of PCNA and CD3 were significantly lower in HJF group than those in model group(P<0.001). MVD was significantly lower in HJF group than that in model group(P<0.05). The mRNA expressions of inflammatory factors(IL-17, IL-23) were significantly lower in HJF group than those in HF group(P<0.05). Conclusion Huoxue Jiedu Fang can inhibit blood vessel proliferation, regulate IL-23/IL-17 axis, reduce expressions of Th17-related factors and relive proliferation of epidermal keratinocytes and infiltration of dermis immunocytes, and thereby relieve the psoriasiform lesions induced by imiquimod in mice.
Objective To observe the interventional effect of Huoxue Jiedu Fang(Blood-activating Toxin-removing Formula) on skin lesions in psoriasiform mouse model induced by imiquimod, and clarify the importance of detoxification Chinese medicinals in treatment of psoriasis. Methods Male BALB/c mice(n =55) were randomly divided into normal control group, model group, Huoxue Jiedu Fang group(HJF group), Huoxue Fang(Blood-activating Formula) group(HF group) and methotrexate group(MTX group, each n =11). The psoriasiform mouse model was induced by external coating imiquimod. MTX group was orally given MTX(1 mg/kg), HJF group, HJF(35.1 g/kg), HF group, HF(26.9 g/kg), while normal control group and model group, equivalent pure water. All mice were executed on the 7~(th) d for collecting skin tissue samples. The scores of psoriasis area and severity index(PASI) were observed. The histomorphological changes and epidermal layer thickness of skin lesion tissue were observed by using light microscope after HE staining. The expressions of proliferating cell nuclear antigen(PCNA), marker on T-cell(CD3) and positive microvessel marker(CD31), and microvessel density(MVD) were detected by using immunohistochemistry technique. The mRNA expressions of interleukin-17(IL-17) and interleukin-23(IL-23) were detected by using real-time PCR. Results The results of HE staining showed that epidermal proliferation, para-keratosis cells and epidermal thickness decreased in HJF group compared with model group(P<0.001). The positive expressions of PCNA and CD3 were significantly lower in HJF group than those in model group(P<0.001). MVD was significantly lower in HJF group than that in model group(P<0.05). The mRNA expressions of inflammatory factors(IL-17, IL-23) were significantly lower in HJF group than those in HF group(P<0.05). Conclusion Huoxue Jiedu Fang can inhibit blood vessel proliferation, regulate IL-23/IL-17 axis, reduce expressions of Th17-related factors and relive proliferation of epidermal keratinocytes and infiltration of dermis immunocytes, and thereby relieve the psoriasiform lesions induced by imiquimod in mice.
引文
[1] Furue M,Kadono T.Psoriasis:Behind the scenes[J].Journal of Dermatology,2016,43(1):4-8.
[2] Boehncke WH,Mrowletz U.Psoriasis[J].Der Hautarzt,2012,63(3):176-177.
[3] Danilenko DM.Review paper:preclinical models of psoriasis[J].vet Pathol,2008,45(4):563-575.
[4] Folkman J.Angiogenesis in psoriasis:therapeutic implications[J].J Invest Dermatol,1972,59(1):40-43.
[5] Weiss MM,Harmange JC,Polverino AJ,et al.Evaluation of a series of naphthamides as potent,orally active vascular en dothelial growth factor receptor-2 tyrosine kinase inhibitors[J].J Med Chem,2008,51(6):1668-1680.
[6] Henno A,Blacher S,Lambert C,et al.Altered expression of angiogenesis and lymph angiogenesis markers in the unin volved skin of plaque-type psoriasis[J].British Journal of Dermatology,2009,160(3):581-590.
[7] Bhushan M,McLaughlin B,Weiss JB,et al.Levels of endothelial cell stimulating angiogenesis factor and vascular endothelial growth factor are elevated in psoriasis[J].Br J Dermatol,1999,141(6):1054-1060.
[8] Heidenreich R,Rocken M,Ghoreschi K.Angiogenesis drives psoriasis pathogenesis[J].Int J Exp Path,2009,90(3):232 -248.
[9] Guttmanyassky E,Krueger JG,Lebwohl MG.Systemic immune mechanisms in atopic dermatitis and psoriasis with im plications for treatment[J].Experimental Dermatology,2017,27(4):409-417.
[10] Andersson AM,Skov L,Thyssen JP,et al.Update on comor bidities in psoriasis[J].Curr Derm Rep,2017,6(2):129-136.
[11] Liu JH,Chen Y,Zhen Z,et al.Relation between endothelial progenitor cells and arterial stiffness in patients with psoriasis[J].Journal of Dermatology,2016,43(8):888-893.
[12] Furue M,Kadono T.“Inflammatory skin march” in atopic der matitis and psoriasis[J].Inflamm Res,2017,66(10):833-842.
[13] 李田田,王萍,周冬梅,等.活血解毒汤治疗银屑病血瘀证的临床疗效观察[J].实用皮肤病学杂志,2016,9(2):133-135.Li TT,Wang P,Zhou DM,et al.Clinical observation of Huoxue Jiedu Decoction for psoriasis vulgaris of blood stasis syn drome[J].Journal of Practical Dermatology,2016,9(2):133 -135.
[14] 张齐雄,曹蓓.中药拳参生物活性研究进展[J].亚太传统医药,2012,8(7):195-196.Zhang QX,Cao B.Progress on bioactivities studies in Chinese native medicine Polygonum Bistorta L[J].Asia-Pacific Traditional Medicine,2012,8(7):195-196.
[15] 高红瑾,陆姗姗.白花蛇舌草乙醇提取物抗炎作用研究[J].中国现代药物应用,2017,11(15):195-196.Gao HJ,Lu SS.Study on the anti-inflammatory effect of ethanol extract of Hedyotis diffusa Willd[J].Chin J Mod Drug Appl,2017,11(15):195-196.
[16] 王生.活血化瘀中药对寻常性银屑病患者白介素-18及尿酸影响的大样本研究[J].中国中医基础医学杂志,2015,21(8):982-984.Wang S.A large sample study effect of Huoxue Huayu De coction on the interleukin-18 and uric acid of patients with psoriasis vulgaris[J].Journal of Basic Chinese Medicine,2015,21(8):982-984.
[17] 高冲,刘璐,胡爱菊,等.活血化瘀中药的药理作用研究进展[J].药学评价研究,2013,36(1):64-68.Gao C,Liu L,Hu AJ,et al.Research progress in pharmaco logical study on Chinese materia medica with function of ac tivating blood and resolving stasis[J].Drug Evaluation Rese arch,2013,36(1):64-68.
[18] Cai YH,Fleming C,Yan J.New insights of T cells in the pathogenesis of psoriasis[J].Cellular & Molecular Immunology,2012,9(4):302-309.
[2] Boehncke WH,Mrowletz U.Psoriasis[J].Der Hautarzt,2012,63(3):176-177.
[3] Danilenko DM.Review paper:preclinical models of psoriasis[J].vet Pathol,2008,45(4):563-575.
[4] Folkman J.Angiogenesis in psoriasis:therapeutic implications[J].J Invest Dermatol,1972,59(1):40-43.
[5] Weiss MM,Harmange JC,Polverino AJ,et al.Evaluation of a series of naphthamides as potent,orally active vascular en dothelial growth factor receptor-2 tyrosine kinase inhibitors[J].J Med Chem,2008,51(6):1668-1680.
[6] Henno A,Blacher S,Lambert C,et al.Altered expression of angiogenesis and lymph angiogenesis markers in the unin volved skin of plaque-type psoriasis[J].British Journal of Dermatology,2009,160(3):581-590.
[7] Bhushan M,McLaughlin B,Weiss JB,et al.Levels of endothelial cell stimulating angiogenesis factor and vascular endothelial growth factor are elevated in psoriasis[J].Br J Dermatol,1999,141(6):1054-1060.
[8] Heidenreich R,Rocken M,Ghoreschi K.Angiogenesis drives psoriasis pathogenesis[J].Int J Exp Path,2009,90(3):232 -248.
[9] Guttmanyassky E,Krueger JG,Lebwohl MG.Systemic immune mechanisms in atopic dermatitis and psoriasis with im plications for treatment[J].Experimental Dermatology,2017,27(4):409-417.
[10] Andersson AM,Skov L,Thyssen JP,et al.Update on comor bidities in psoriasis[J].Curr Derm Rep,2017,6(2):129-136.
[11] Liu JH,Chen Y,Zhen Z,et al.Relation between endothelial progenitor cells and arterial stiffness in patients with psoriasis[J].Journal of Dermatology,2016,43(8):888-893.
[12] Furue M,Kadono T.“Inflammatory skin march” in atopic der matitis and psoriasis[J].Inflamm Res,2017,66(10):833-842.
[13] 李田田,王萍,周冬梅,等.活血解毒汤治疗银屑病血瘀证的临床疗效观察[J].实用皮肤病学杂志,2016,9(2):133-135.Li TT,Wang P,Zhou DM,et al.Clinical observation of Huoxue Jiedu Decoction for psoriasis vulgaris of blood stasis syn drome[J].Journal of Practical Dermatology,2016,9(2):133 -135.
[14] 张齐雄,曹蓓.中药拳参生物活性研究进展[J].亚太传统医药,2012,8(7):195-196.Zhang QX,Cao B.Progress on bioactivities studies in Chinese native medicine Polygonum Bistorta L[J].Asia-Pacific Traditional Medicine,2012,8(7):195-196.
[15] 高红瑾,陆姗姗.白花蛇舌草乙醇提取物抗炎作用研究[J].中国现代药物应用,2017,11(15):195-196.Gao HJ,Lu SS.Study on the anti-inflammatory effect of ethanol extract of Hedyotis diffusa Willd[J].Chin J Mod Drug Appl,2017,11(15):195-196.
[16] 王生.活血化瘀中药对寻常性银屑病患者白介素-18及尿酸影响的大样本研究[J].中国中医基础医学杂志,2015,21(8):982-984.Wang S.A large sample study effect of Huoxue Huayu De coction on the interleukin-18 and uric acid of patients with psoriasis vulgaris[J].Journal of Basic Chinese Medicine,2015,21(8):982-984.
[17] 高冲,刘璐,胡爱菊,等.活血化瘀中药的药理作用研究进展[J].药学评价研究,2013,36(1):64-68.Gao C,Liu L,Hu AJ,et al.Research progress in pharmaco logical study on Chinese materia medica with function of ac tivating blood and resolving stasis[J].Drug Evaluation Rese arch,2013,36(1):64-68.
[18] Cai YH,Fleming C,Yan J.New insights of T cells in the pathogenesis of psoriasis[J].Cellular & Molecular Immunology,2012,9(4):302-309.