苯烯莫德对实验性银屑病的治疗作用
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摘要
目的观察苯烯莫德对实验性银屑病的影响。方法体外培养永生化人角质生成细胞Ha Ca T,用MTT法测试苯烯莫德的抑制率,免疫组化法观察角蛋白19(CK19)的表达。体内试验以维A酸乳膏为阳性对照,观察0.5%、1%、2%苯烯莫德药效(苯烯莫德低、中、高剂量组),并设阴性对照组。雌性性成熟小鼠60只,雌二醇诱导小鼠动情期阴道上皮有丝分裂模型成功后随机分组,从阴道注入样品(50μL),检测阴道组织有丝分裂指数。60只小鼠随机分组,给药前各组动物均用尼龙刷轻刷尾根0.5 cm处100次制作小鼠尾部颗粒层鳞片数模型,然后在同一部位涂抹给药(100μL),计数给药部位每100个鳞片中有颗粒层的鳞片数。80只豚鼠随机分组,普萘洛尔诱导豚鼠耳背部银屑病样模型成功后涂抹药剂(约200μL)于豚鼠耳背,取给药部位皮肤显微测表皮厚度。结果体外试验显示苯烯莫德对Ha Ca T细胞的IC50为28.5μg·m L-1,Ha Ca T细胞在20μg·m L-1苯烯莫德中培养,有大量CK19生成。苯烯莫德能抑制小鼠阴道上皮细胞的有丝分裂,苯烯莫德各剂量组有丝分裂细胞指数低于赋形剂组(P<0.05或P<0.01),苯烯莫德中、高剂量组低于苯烯莫德低剂量组(P<0.05,P<0.01)。苯烯莫德处理后能增加含颗粒层的鳞片数,苯烯莫德各剂量组小鼠尾部有颗粒层的鳞片比率高于赋形剂组(P<0.05或P<0.01),苯烯莫德中、高剂量组高于苯烯莫德低剂量组(P<0.05,P<0.01)。普萘洛尔诱导的豚鼠耳背部表皮的厚度增加能够被苯烯莫德抑制,苯烯莫德各剂量组表皮厚度薄于赋形剂组(P<0.05或P<0.01),苯烯莫德高剂量组薄于苯烯莫德低剂量组(P<0.05)。结论苯烯莫德具有明显治疗实验性银屑病的作用,具有开发前景。
AIM To investigate the ameliorative effects of benvitimod on experimental psoriasis in vitro and in vivo. METHODS The effects of benvitimod on Ha Ca T cell survival were measured by MTT, and the keratin 19 protein(CK19) expression of Ha Ca T cell were detected by immunohistochemical staining. The effects of 0.5%, 1% and 2% benvitimod( benvitimod low, middle and high dose group) on three psoriasis animal models were investigated and compared with negative control group and vitamin A acid cream positive control group. First, 60 mature female mice were induced by estradiol injection to establish the animal model of vaginal tissues mitosis, and the effect of benvitimod on vaginal epithelium mitotic index was evaluated. Second, a mouse tail granular layer scale model was induced by brushing on the mice tail root for 100 times before benvitimod treating, then counting the number of particles in each of the 100 scales of the applicator parts.Third, ears back psoriasis model was induced by using 5% propranolol on 80 guinea pigs, then applying benvitimod on the same parts and the epidermis thickness was measured. RESULTS The IC50 of benvitimod on Ha Ca T cells was 28.5 μg·m L-1. There was much of CK19 expression of Ha Ca T cell while treated with 20 μg·m L-1benvitimod. The number of mitotic cells in mouse vaginal epithelial cells was significantly inhibited by benvitimod. The mitotic cell rates of benvitimod high, middle and low dose group were less than the excipient group( P < 0.05 or P < 0.01), and the benvitimod middle and high dose group were less than the low dose group(P < 0.05, P < 0.01). The number of mice granular layer scales was significantly increased in benvitimod high, middle and low dose group compared with the excipient group( P < 0.05 or P < 0.01), and the benvitimod middle and high dose group were more than the low dose group(P < 0.05, P < 0.01). The increased ears epidermis thickness in propranolol- induced mice was significantly inhibited by benvitimod. The epidermis thickness of benvitimod high, middle and low dose group were thinner compared with the excipient group(P <0.05 or P < 0.01), and the benvitimod high dose group was thinner than the low dose group( P < 0.05).CONCLUSION Benvitimod engages the ameliorative effects on experimental psoriasis and may be as a potential new anti- psoriasis drug of topical application.
AIM To investigate the ameliorative effects of benvitimod on experimental psoriasis in vitro and in vivo. METHODS The effects of benvitimod on Ha Ca T cell survival were measured by MTT, and the keratin 19 protein(CK19) expression of Ha Ca T cell were detected by immunohistochemical staining. The effects of 0.5%, 1% and 2% benvitimod( benvitimod low, middle and high dose group) on three psoriasis animal models were investigated and compared with negative control group and vitamin A acid cream positive control group. First, 60 mature female mice were induced by estradiol injection to establish the animal model of vaginal tissues mitosis, and the effect of benvitimod on vaginal epithelium mitotic index was evaluated. Second, a mouse tail granular layer scale model was induced by brushing on the mice tail root for 100 times before benvitimod treating, then counting the number of particles in each of the 100 scales of the applicator parts.Third, ears back psoriasis model was induced by using 5% propranolol on 80 guinea pigs, then applying benvitimod on the same parts and the epidermis thickness was measured. RESULTS The IC50 of benvitimod on Ha Ca T cells was 28.5 μg·m L-1. There was much of CK19 expression of Ha Ca T cell while treated with 20 μg·m L-1benvitimod. The number of mitotic cells in mouse vaginal epithelial cells was significantly inhibited by benvitimod. The mitotic cell rates of benvitimod high, middle and low dose group were less than the excipient group( P < 0.05 or P < 0.01), and the benvitimod middle and high dose group were less than the low dose group(P < 0.05, P < 0.01). The number of mice granular layer scales was significantly increased in benvitimod high, middle and low dose group compared with the excipient group( P < 0.05 or P < 0.01), and the benvitimod middle and high dose group were more than the low dose group(P < 0.05, P < 0.01). The increased ears epidermis thickness in propranolol- induced mice was significantly inhibited by benvitimod. The epidermis thickness of benvitimod high, middle and low dose group were thinner compared with the excipient group(P <0.05 or P < 0.01), and the benvitimod high dose group was thinner than the low dose group( P < 0.05).CONCLUSION Benvitimod engages the ameliorative effects on experimental psoriasis and may be as a potential new anti- psoriasis drug of topical application.
引文
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[8]孙悦,石玉,魏波,等.银屑病治疗的小分子药物研究进展[J].现代药物与临床,2014,29(11):1318-1325.
[9]陈永锋.靶向生物制剂治疗银屑病的免疫学机制和应用[J].皮肤性病诊疗学杂志,2015,22(4):271-273.
[10]胡雨来,吴江林.(e)—3,5-二甲氧基-4’-乙酰氧基二苯乙烯的制备方法:中国,CN201310225535[P].2013-08-28.
[11]ZHAO L,CHEN X,CAI L,et al.Randomized,double-blind,placebo-controlled,multiple-dose study of the safety,tolerability and pharmacokinetics of benvitimod,a candidate drug for the treatment of psoriasis[J].J Clin Pharm Ther,2014,39(4):418-423.
[12]杨素清,闫景东,王松岩.蜈蚣败毒饮对雌激素周期小鼠阴道上皮细胞有丝分裂的影响[J].中医药学报,2010,38(5):33-35.
[13]刘晋华,李玉萍,尤光甫.青黛克银丸治疗银屑病模型的实验研究[J].中国实验方剂学杂志,2010,16(16):126-128.
[14]伍长娟,沈娟.盐酸普萘洛尔造模周期对豚鼠银屑病模型的影响[J].承德医学院学报,2015,32(5):374-376.
[15]孙林潮.银屑病动物模型研究进展[J].国外医学皮肤性病学分册,1995,21(5):264-266.
[16]KRANING KK.Research needs in 11 major areas in dermatology.I.Psoriasis[J].J Invest Dermatol,1979,73(5 Pt 2):402-413.
[17]GAYLARDE PM,BROCK AP,SARKANY I.Psoriasiform changes in guinea-pig skin from propranolol[J].Clin Exp Dermatol,1978,3(2):157-160.
[18]THAKKAR K,GEAHLEN RL,CUSHMAN M.Synthesis and protein-tyrosine kinase inhibitory activity of polyhydroxylated stilbene analogues of piceatannol[J].J Med Chem,1993,36(20):2950-2955.
[19]陈庚辉,约翰·麦尔康姆·韦伯斯特.用羟基芪、新型芪衍生物及其类似物抗发炎、治疗牛皮癣和抑制蛋白质致活酶:中国,CN00816755[P].2003-04-02.
[2]张建中.银屑病的流行病学与危险因素[J].实用医院临床杂志,2013,10(1):4-6.
[3]ENAMANDRAM M,KIMBALL AB.Psoriasis epidemiology:the interplay of genes and the environment[J].J Invest Dermatol,2013,33(2):287-289.
[4]OLIVEIRA MDE F,ROCHA BDE O,DUARTE GV.Psoriasis:classical and emerging comorbidities[J].An Bras Dermatol,2015,90(1):9-20.
[5]张晓红.银屑病的临床表现与诊断[J].中国临床医生,2009,37(8):8-10.
[6]张培,单葵.银屑病外用药物治疗进展[J].中国药房,2015,26(17):2446-2448.
[7]WEINBERG JM,SAINI R,TUTRONE WD.Biologic therapy for psoriasis—the first wave:infliximab,etanercept,efalizumab,and alefacept[J].J Drugs Dermatol,2002,1(3):303-310.
[8]孙悦,石玉,魏波,等.银屑病治疗的小分子药物研究进展[J].现代药物与临床,2014,29(11):1318-1325.
[9]陈永锋.靶向生物制剂治疗银屑病的免疫学机制和应用[J].皮肤性病诊疗学杂志,2015,22(4):271-273.
[10]胡雨来,吴江林.(e)—3,5-二甲氧基-4’-乙酰氧基二苯乙烯的制备方法:中国,CN201310225535[P].2013-08-28.
[11]ZHAO L,CHEN X,CAI L,et al.Randomized,double-blind,placebo-controlled,multiple-dose study of the safety,tolerability and pharmacokinetics of benvitimod,a candidate drug for the treatment of psoriasis[J].J Clin Pharm Ther,2014,39(4):418-423.
[12]杨素清,闫景东,王松岩.蜈蚣败毒饮对雌激素周期小鼠阴道上皮细胞有丝分裂的影响[J].中医药学报,2010,38(5):33-35.
[13]刘晋华,李玉萍,尤光甫.青黛克银丸治疗银屑病模型的实验研究[J].中国实验方剂学杂志,2010,16(16):126-128.
[14]伍长娟,沈娟.盐酸普萘洛尔造模周期对豚鼠银屑病模型的影响[J].承德医学院学报,2015,32(5):374-376.
[15]孙林潮.银屑病动物模型研究进展[J].国外医学皮肤性病学分册,1995,21(5):264-266.
[16]KRANING KK.Research needs in 11 major areas in dermatology.I.Psoriasis[J].J Invest Dermatol,1979,73(5 Pt 2):402-413.
[17]GAYLARDE PM,BROCK AP,SARKANY I.Psoriasiform changes in guinea-pig skin from propranolol[J].Clin Exp Dermatol,1978,3(2):157-160.
[18]THAKKAR K,GEAHLEN RL,CUSHMAN M.Synthesis and protein-tyrosine kinase inhibitory activity of polyhydroxylated stilbene analogues of piceatannol[J].J Med Chem,1993,36(20):2950-2955.
[19]陈庚辉,约翰·麦尔康姆·韦伯斯特.用羟基芪、新型芪衍生物及其类似物抗发炎、治疗牛皮癣和抑制蛋白质致活酶:中国,CN00816755[P].2003-04-02.