【作者】 陈旭娥；

【导师】 李家文；

【作者基本信息】 华中科技大学 ， 皮肤病与性病学， 2008， 博士

【摘要】 目的:(1)研究扁平苔藓临床病理特征,为进一步探讨扁平苔藓的发病机制及指导其治疗提供依据;(2)探讨炎性细胞因子(TNF-α、ICAM-1)、凋亡相关蛋白(Caspase-3、Bax)、细胞周期蛋白(Cyclin D1)及Th1型细胞因子(IL-12)在扁平苔藓患者中的表达及临床意义。方法:(1)回顾2005年1月至2007年12月在华中科技大学同济医学院附属协和医院皮肤科经临床、病理确诊为扁平苔藓患者83例,结合所选研究对象的临床资料和病理资料,按不同条件分组,进行临床和病理资料分析;(2)采用免疫组化SP法对29例扁平苔藓患者皮损组织和10正常人皮肤组织中TNF-α、ICAM-1、Caspase-3、Bax、Cyclin D1蛋白表达水平进行检测;(3)采用逆转录聚合酶链反应(RT-PCR)方法对29例扁平苔藓患者皮损组织和10例正常人皮肤组织中IL-12p40mRNA、IL-12p35mRNA的表达水平进行检测;(4)分离29例扁平苔藓患者和10例正常人外周血单个核细胞(PBMC),检测PBMC培养上清液中IL-12蛋白水平。结果:(1)83例扁平苔藓患者临床病理分析结果①一般资料:发病男女无性别差异(χ2=0.11,P>0.05),男女比例为1.08:1,平均年龄为42.84±15.97岁;以30-39岁年龄之间最多;平均病程33.54月;病程<1年组与≥1年组比较,差异无统计学意义(χ2=0.51,P>0.05);②发病部位以两处或两处以上同时受累最多见,占34.94%;其次为口腔和四肢受累,各占19.28%;躯干和臀、腹股沟、外生殖器受累各占12.05%;面部受累最少,占2.40%。组间比较,差异有统计学意义(χ2=29.55,P<0.05);③皮损以紫红色、紫褐色丘疹、斑片者最多,占61.45%;以糜烂、溃疡为主者占12.05%;灰黑色、灰青色、灰褐色丘疹、斑片为主者占7.23%;其它少见非典型皮疹占19.27%。组间比较,差异有统计学意义(χ2=61.24,P <0.05);④67.47%的患者无自觉症状或症状轻微,不影响正常工作和学习,27.71%的患者诉严重瘙痒,4.82%患者疼痛症状明显。组间比较,差异有统计学意义(χ2=50.05,P<0.05);⑤100%病例病理象示基底细胞液化变性和真皮浅层炎症细胞浸润;54.22%病例见胶样小体形成;63.86%的病例存在明显表皮病理学改变;3.61%出现表皮下裂隙或水疱;⑥首诊即靠临床特点即可确诊扁平苔藓44例,仅占53.10%。(2)TNF-α蛋白在扁平苔藓皮损组织和正常人皮肤组织中的表达29例扁平苔藓皮损组织中,TNF-α阳性表达率为93.10%,10例正常人皮肤组织中TNF-α阳性表达率仅为30.00%,组间比较差异有统计学意义(χ2=19.40,P<0.05)。(3)ICAM-1在扁平苔藓皮损组织和正常人皮肤组织中的表达29例扁平苔藓皮损组织中,ICAM-1阳性表达率为89.66%,10例正常人皮肤组织均未检测到ICAM-1阳性表达,阳性率为0.00%,组间比较,差异有统计学意义(χ~2=26.90,P<0.05)。(4)Caspase-3在扁平苔藓皮损组织和正常人皮肤组织中的表达29例扁平苔藓皮损组织中,Caspase-3阳性表达率为86.20%,10例正常皮肤组织中,Caspase-3阳性表达率为20.00%,组间比较,差异有统计学意义(χ2=14.15,P<0.05)。(5)Bax在扁平苔藓皮损组织和正常人皮肤组织中的表达29例扁平苔藓皮损组织中,Bax阳性表达率为93.10%,10例正常人皮肤组织中,Bax阳性表达率为50.00%,组间比较,差异有统计学意义(χ2=17.60,P<0.05)。(6)Cyclin D1在在扁平苔藓皮损组织和正常人皮肤组织中的表达29例扁平苔藓皮损组织中,Cyclin D1阳性表达率为79.31%,10例正常人皮肤组织中,Cyclin D1表达阳性率为30.00%,组间比较,差异有统计学意义(χ2=8.57,P<0.05)。(7)IL-12p40mRNA在扁平苔藓皮损组织和正常人皮肤组织中的表达29例扁平苔藓患者皮损处均表达IL-12p40mRNA,而10例正常皮肤组织均不能检测到IL-12p40mRNA,组间比较差异均有统计学意义(F=9.03,P<0.05)。(8)IL-12p35mRNA在扁平苔藓皮损组织和正常人皮肤中的表达29例扁平苔藓患者皮损组织和10例正常人皮肤组织中均表达IL-12p35 mRNA,但皮损组和正常对照组组间比较,无显著性差异(F=3.71,P>0.05)。(9)IL-12蛋白在扁平苔藓患者和正常人外周血PBMC培养上清中的检测29例扁平苔藓患者外周血PBMC培养上清液IL-12平均水平为57.14±19.64pg/mL,而10例正常对照组外周血PBMC培养上清IL-12平均水平为30.10±7.45pg/mL。两者比较,差异有统计学意义(F=5.75,P<0.05)。结论:(1)83例扁平苔藓患者以30-39岁年龄组发病最多;(2)发病部位以两处或两处以上同时受累居多;临床表现多样,以紫红色、紫褐色丘疹、斑片者最多;患者多无自觉症状或症状轻微,不影响正常工作和学习;(3)扁平苔藓病理特征为基底细胞液化变性和真皮浅层炎症细胞浸润;部分病例可见胶样小体形成、表皮病理学改变等;(4)必须临床与病理相结合才能正确诊断扁平苔藓;(5)TNF-α、ICAM-1在扁平苔藓皮损局部异常高表达,可能促进皮损局部慢性炎症发应,与扁平苔藓发病有关;(6)Caspase-3、Bax在扁平苔藓皮损局部异常高表达,可能促进皮损局部角质形成细胞凋亡,与扁平苔藓发病有关;(7)Cyclin D1在扁平苔藓皮损局部异常高表达,可能促进角质形成细胞细胞周期的进程,促进其增殖,可能有助于维持表皮形态的完整性,在机体抗扁平苔藓过程中发挥保护性作用;(8)IL-12p40mRNA在扁平苔藓皮损局部异常高表达和外周血PBMC分泌IL-12增高,可能与扁平苔藓发病过程中Th1型自身免疫反应有关。更多还原

【Abstract】 Objective:(1)To study the clinical feature and pathologic character of lichen planus.(2) To investigate the role of proinflammation cytokine (TNF-αand ICAM-1), apoptosis related protein (Caspase-3 and Bax), Cyclin D1, and Th1-type cytokine (IL-12) in the pathogenesis of lichen planus.Methods:(1) 83 lichen planus patients from Union hospital of Huazhong University of Science Technology which had been diagnosed by clinical character and skin biopsy between Jan 2005 and Dec 2007 were reviewed;(2) Immunohistochemical technique was employed to observe the expression of TNF-α, ICMA-1, Caspase-3, Bax and Cyclin D1 in the lesional skin from 29 patients with lichen planus. Normal skin from 10 healthy persons was used as control;(3) The expressions of IL-12p35 and IL-12p40 mRNA were semi-quantitatively analyzed with reverse transcriptase-polymerase chain reaction (RT-PCR) in the lesional skin from 29 patients with lichen planus and 10 healthy persons.(4) PBMC was obtained from 29 patients with LP and 10 normal controls. IL-12 in the culture supernatants were measured quantitatively by ELISA.Results:(1) According to our information, the onset of LP generally occurred in the middle-aged especially 30-39 years old (mean age 42.84±15.97), with the ratio of male to female 1.08:1;(2) The lesions always involved more than one part of body. The clinical character is diversifying, and prunosus papular and patch were the most clinical manifestations. Many patients were asymptomatic or symptomatic in low degree;(3) The histopathology of LP is the subepithelial band-like inflammatory cell infiltrate predominated by lymphocytes, and destruction of the epithelial basal cell layer. In some cases, there is Civatte bodies and change in epidermal skin;(4) Only depend on clinical character, the dermatologist can not diagnosis lichen planus correctly;(5) Positive TNF-αexpressions rate in lichen planus were significantly higher than those in normal skins (χ~2=19.40,P<0.05);(6) Positive ICAM-1 expressions rate in lichen planus were significantly higher than those in normal skins (χ~2=26.90,P<0.05);(7) Positive Caspase-3 expressions rate in lichen planus were significantly higher than those in normal skins (χ~2=14.15,P<0.05);(8) Positive Bax expressions rate in lichen planus were significantly higher than those in normal skins (χ~2=17.60,P<0.05);(9) Positive Cyclin D1 expressions rate in lichen planus were significantly higher than those in normal skins (χ~2=8.57,P<0.05);(10) The lever of IL-12p40mRNA in lichen planus were significantly higher than those in normal skins (F=9.03,P<0.05);(11) There is no difference in the lever of IL-12p35mRNA of lichen planus and normal skins (F=3.71,P>0.05);(12) IL-12 in the culture supernatants of PBMC from patients with lichen planus were significantly higher than those from normal controls (F=5.75,P<0.05).Conclusions:(1) According to our information, the onset of LP generally occurred in the middle-aged especially 30-39 years old (mean age 42.84±15.97), with the ratio of male to female 1.08:1; (2) The lesions always involved more than one part of body. The clinical character is diversifying, and prunosus papular and patch were the most clinical manifestations. Many patients were asymptomatic or symptomatic in low degree;(3) The histopathology of LP is the subepithelial band-like inflammatory cell infiltrate predominated by lymphocytes, and destruction of the epithelial basal cell layer. In some cases, there is Civatte bodies and change in epidermal skin;(4) In order to diagnosis lichen planus correctly, the dermatologist must master the clinical and pathologic character of LP;(5) The expression of TNF-αand ICAM-1 might play an important role in the chronic inflammation of lichen planus;(6) The expression of Caspase-3 and Bax might play an important role in the keratinocyte apoptosis of lichen planus;(7) The expression of Cyclin D1 might play an important role in the proliferation of keratinocyte in lichen planus;(8) The overexpression of IL-12p40mRNA in lesional skin and IL-12 in the culture supernatants of PBMC might play an important role in the Th1-type autoimmunologic reaction of LP.更多还原