Enhancing effect of tazarotene on the HLA-DR expression of cultured human keratinocytes induced by interferon-gamma

Objective: To investigate the effect of tazarotene on the expression of HLA-DR induced by IFN-γ. Methods: (1) Keratinocytes from normal human skin were cultured in vitro;(2) Tazarotene, IFN-γ and the combination of the two compounds were incubated with the keratinocytes in medium, respectively. The expression of HLA-DR in keratinocytes was determined using immunocytochemistry techniques at 24h after incubation. Results: (1) There was rare expression of HLA-DR in normal human keratinocytes; (2) 10-6mol/L tazarotene failed to induce the expression of HLA-DR in keratinocytes at 24h after incubation; (3) 500 U/ml IFN-γ obviously induced the HLA-DR expression in keratinocytes at 24h after treatment; (4) After 24h, 10-7-10-5 mol/L tazarotene had a significantly enhancing effect on the expression of HLA-DR induced by IFN-γ (P<0.005). Conclusion: Tazarotene up-regulates the expression of HLA-DR in keratinocytes cultured in vitro when combined with IFN-γ . Therefore, the reduction of HLA-DR positive keratinocytes in psoriatic lesions may be attributed to not direct interaction of tazarotene in combination with IFN-γ but other pathways.     

The expression of Fas, FasL and their biological behavior in human cervical carcinoma

Objective： To investigate the relationship between the expression of Fas and Fas ligand （FasL） and its biological behavior in human cervix carcinoma. Methods： Immunohistochemisty technique was used to detect the expression of Fas and FasL in 47 cases of cervical carcinoma, 16 cases of cervical interaepithelial neoplasia, 10 cases of chronic cervicitis and 10 cases of normal cervix. TUNEL technique was used to observe the apoptic cells in 47 cases of cervical carcinoma. Retrospective study was carried out to find the relationship between the expression of Fas and FasL and cell apoptosis, clinical stage, pathological classification, lymph node metastasis, prognosis and age. Results： The expression of Fas and FasL was significantly different in different cervix （P 〈 0.01）, and also related to the degree of differentiation, lymph node metastasis and prognosis （P 〈 0.05）. But had no relation with clinical stage or age （P 〉 0.05） ; Cervix carcinoma cells apoptosis in different pathological classification appeared negative relation （Rs=-0.35, P 〈 0.05 ）. Cervix carcinoma cell apoptosis was significantly higher in Fas-positive and FasL- positive than that in Fas-negative and FasL-negative （P 〈 0.05）. By retrospective investigation, Fas-negative and FasL-positive were related to poor prognoses of the patients with cervical carcinoma （P 〈 0.05 ）. Conclusion： The development of apoptosis in cervix carcinoma has a promoting regulation function in Fas and FasL expression. Gene treatment can alter apoptosis abnormality, thus induce apoptosis in cancerous cell expressing Fas and FasL. Fas or FasL may be taken as a marker in the prognostic character- ization.     

Expression of IRF-4 and IBP in skin lesions of patients with psoriasis vulgaris

The expression of the interferon regulatory factor 4 (IRF-4) and the IRF-4-binding pro-tein (IBP) in psoriatic skin lesions was investigated. The expression of IRF-4 and IBP in skin lesions of 20 patients with psoriasis vulgaris were immunohistochemically dectected. Normal skin from 10 healthy people was used as normal control. The study showed that expression of IRF-4 was increased significantly in keratinocytes and inflammatory cells in the lesions of psoriasis vulgaris than that in the normal control. The detection revealed that IBP expression in keratinocytes, lymphocytes, hair follicles, and sebaceous glands in normal skin was significantly lower than that in the lesions of pso-riasis vulgaris (P<0.05). Both IRF-4 and IBP might be involved in the pathogenesis of psoriasis vul-garis.     

Study on Vip protein expression in psoriatic epidermis with the topical treatment of capsaicin ointment

Objective： To investigate the mechanism of capsaicin in treating active psoriasis vulgaris. Methods： VIP protein in active psoriatic lesions before and 30 days after the treaanent of capsaicin ointment was detected by immunohistochemistry. Results： There was positive expression of VIP in all layers of psoriatic lesions epidermis （95.5 % ）, but after the treatment of capsaicin ointment, there was nearly no expression of VIP protein in epidermis（22.2% ）. Conclusion： Capsaicin inhibits proliferation and induces the differentiation of keratinocytes through down-regulating the expression of VIP in psoriatic epidermis.     

Anti-Fas antibody induces apoptosis in cultured human renal interstitial fibroblasts

Objective To detect if Fas is expressed in human renal interstitial fibroblasts (hRIFs) an d apoptosis of hRIFs can be induced by specific anti-Fas antibody.Methods hRIFs were cultured from isolated papillae of human kidney, and identified by mo rphologic examination, assay of antigenic components and culture in D-valine se lective medium.Fas expression in normal hRIFs was detected by RT-PCR and imm unocytochemistry staining.After hRIFs were incubated with interferon gamma (γ -IFN 500 U/ml, 1000 U/ml, 1500 U/ml and 2000 U/ml, respectively) for 48 hou rs, Fas expression was determined by Northern blot, Western blot and flow cytome try.hRIFs pre-stimulated with γ-IFN (500 U/ml, 48 hours) were incubated wi th anti-Fas antibody (IgM, 0.5 μg/ml) for 12 hours.And apoptosis was ident ified by morphologic examination, DNA ladder assay and flow cytometry. Results The cultured hRIFs showed a shuttle-like shape and were positively stained by l abeled anti-vimentin antibody but negatively stained by anti-epithelial membra ne antibody.They could not grow in the D-valine selective medium and died par tly in a week.Fas mRNA and protein were expressed in normal hRIFs and markedly upregulated by stimulation with γ-IFN.Apoptosis in γ-IFN pre-stimula ted hRIFs was induced by anti-Fas antibody, showing cell nuclear shrinkage and condensation in morphologic feature, internucleosomal DNA fragmentation in DNA l adder assay and a pick of hypo-diploid nuclei by flow cytometry.Conclusion Fas is normally expressed in hRIFs and can be markedly upregulated by γ-IFN. Anti-Fas antibodies can induce apoptosis of hRIFs pre-stimulated with γ-IF N.     

Effects of the Combined Use of Benazepril and Valsartan on Apoptosis in the Kidney of Rats with Adriamycin-induced Nephritic Gomerulosclerosis

The effects of the combined use of angiotensin converting enzyme inhibitor (ACEI) benazepril and angiotensin 11 type 1 receptor antagonist (AT1RA) valsartan on apoptosis and the expression of apoptosis-related proteins Fas and FasL in the kidney of rats with adriamycin-induced nephritic glomerulosclerosis was investigated. Uninephrectomy and the injection of adriamycin induced the rat model of glomerulosclerosis. Benazepril (6 mg/kg), valsantan (20 mg/kg), or benazepril (3 mg/kg) plus valsantan (20 mg/kg) was respectively delivered daily by gavage to the rats in three treatment groups for 12 weeks. Apoptosis was examined by means of terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL). Immunohistochemistry was adopted to detect the expression of Fas and FasL. Software of pathological analysis quantitated the levels of Fas and FasL. The results showed that as compared with those in the control group, the kidneys in the model group had more severe glomerulosclerosis, much more apoptotic cells and higher levels of expression of Fas and FasL. The degree of glomerulosclerosis, the number of apoptotic cells and the levels of expression of Fas and FasL were reduced by benazepril and valsartan. The combined use of benazepril and valsartan had the best therapeutic effect. It was concluded that benazepril and valsartan could suppress the excessive apoptosis of kidney cells by lowering the expression of the apoptosis-related proteins Fas and FasL, so as to postpone the process of glomerulosclerosis. The combined use of benazeoril and valsartan has better theraoeutic effect.     

Dioscin-induced Apoptosis of Human LNCaP Prostate Carcinoma Cells through Activation of Caspase-3 and Modulation of Bcl-2 Protein Family简

Dioscin is a natural steroid saponin derived from several plants, showing potent anti-cancer effect against a variety of tumor cell lines. In the present study, we investigated the anti-cancer activity of dioscin against human LNCaP cells, and evaluated the possible mechanism involved in its antineoplastic action. It was found that dioscin（1, 2 and 4 μmol/L） could significantly inhibit the viability of LNCaP cells in a time- and concentration-dependent manner. Flow cytometry revealed that the apoptosis rate was increased after treatment of LNCaP cells with dioscin for 24 h, indicating that apoptosis was an important mechanism by which dioscin inhibited cancer. Western blotting was employed to detect the expression of caspase-3, Bcl-2 and Bax in LNCaP cells. The expression of cleaved caspase-3 was significantly increased, and meanwhile procaspase-3 was markedly decreased. The expression of anti-apoptotic protein Bcl-2 was down-regulated, whereas the pro-apoptotic protein Bax was up-regulated. Moreover, the Bcl-2/Bax ratio was drastically decreased. These results suggested that dioscin possessed potential anti-tumor activity in human LNCaP cells through the apoptosis pathway, which might be associated with caspase-3 and Bcl-2 protein family.     

Study of apoptosis induced by nordihydroguaiaretic acid in human ma-lignant glioma cell line SHG-44

Objective: To investigate the effect and mechanism of nordihydroguaiaretic acid (NDGA) on apop-tosis in human malignant glioma cell line SHG-44. Methods: Cell growth inhibition was measured with MTT assay. Cell apoptosis was observed with light and electron microscopy and TUNEL. Expression of bcl-2 gene was measured with immunohistochemistry, in situ hybridization and image analyses. Results: NDGA at the concentration of 100 μmol/L inhibited the proliferation of SHG-44 cells and induced apoptosis in a time-de-pendent manner. The expression of Bcl-2 protein in SHG-44 cells was decreased in the present of 100 μmol/L NDGA along with the duration of treatment in a negative correlation with the degree of cell apoptosis. The bcl-2 mRNA expressed in SHG-44 cells was reduced after treatment with 100 μmol/L NDGA, apparently consistent with the immunohistochemical results. Conclusion.- NDGA can induce apoptosis of human malig-nant glioma cells probably by down-regulating expression of bcl-2 gene, though the exact mechanism needs further study.     

miR-21 targets Fas ligand-mediated apoptosis in breast cancer cell line MCF-7

Over-expression of Fas ligand （FasL） on tumor cell surface can induce the apoptosis of spe- cific activated tumor infiltrating lymphocytes （TILs） via the Fas/FasL pathway, leading to the formation of a site of immune privilege surrounding the tumor mass for escaping immune surveillance and pro- moting tumor proliferation, invasion and metastasis. The blocking effect of miR-21 on FasL-mediated apoptosis in breast cancers was investigated in this study. The expression levels of miR-21 and FasL in human breast carcinoma cell lines were detected by using RT-PCR and Western blotting. FasL as a tar- get gene of miR-21 was identified by Luciferase assay. The apoptosis of Jurkat T lymphocytes induced by MCF-7 cells was determined by flow cytometry. It was found that in four human breast cancer cell lines, FasL expression level in MCF-7 cells was the highest, while miR-21 was down-regulated the most notably. After miR-21 expression in MCF-7 cells was up-regulated, FasL was identified as a target gene of miR-21. When the effector/target （E/T） ratio of MCF-7 cells and Jurkat cells was 10：1, 5：1 and 1：1, the inhibitory rate of apoptosis of Jurkat T lymphocytes induced by MCF-7 cells was 95.81%, 93.16% and 91.94%, respectively. It is suggested that in breast cancers miR-21 expression is negatively associ- ated with FasL expression, and FasL is a target gene of miR-21, miR-21 targeting and regulating FasL-mediated apoptosis will bring us the possibility of a new tumor immunotherapy via breaking tu- mor immune privilege.     

奥曲肽诱导肝癌细胞凋亡和Fas/FasL的表达

目的 探讨奥曲肽对人肝癌细胞抑制作用的机制,为奥曲肽临床应用提供实验依据.方法 用奥曲肽4、7、10 μ g/ml作用人肝癌细胞株(SMMC-7721)24、48、72 h,以空白作为对照组;采用原位末端标记(TUNEL)法流式细胞仪检测SMMC-7721凋亡;直接标记单克隆(CD95/CDl78)抗体法流式细胞仪检测SMMC-7721 Fas/FasL表达的变化.结果 奥曲肽在4、7、10 μg/ml d作用 SMMC-7721 24 h后,SMMC-7721凋亡率从8%升高为40%(P＜0.05),Fas表达率从57%升高为76%(P＜0.05),FasL表达率从54%升高为63%(P＜0.05).结论 奥曲肽能诱导肝癌细胞(Hep SMMC-7721)凋亡的发生,而且能促进肝癌细胞Fas/Fasl基因的表达.     

Factors influencing the expression of Fas and Fas ligand on newborn murine ker atinocytes and fibroblasts

Ｆａｓ (ＣＤ95)ａｎｄＦａｓｌｉｇａｎｄ (ＦａｓＬ) ,ａｓｍｅｍｂｅｒｓｏｆＴＮＦｆａｍｉｌｉｅｓ ,ａｒｅｔｙｐｅⅠａｎｄｔｙｐｅⅡｃｅｌｌｕｌａｒｔｒａｎｓｍｅｍｂｒａｎｅｐｒｏｔｅｉｎｓ ＦａｓＬｉｎｄｕｃｅｓａｐｏｐｔｏｓｉｓｂｙｂｉｎｄｉｎｇｔｏｉｔｓｒｅｃｅｐｔｏｒＦａｓｏｎｔｈｅｃｅｌｌｓｕｒｆａｃｅ Ｔｈｉｓｐｒｏｃｅｓｓｉｓｉｎｖｏｌｖｅｄｉｎｉｍｍｕｎｅｒｅｓｐｏｎｓｅｔｏｖｉｒｕｓ ,ｔｕｍｏｒｃｅｌｌｓａｎｄｔｈｅｒｅｇｕｌａｔｉｏｎｏｆｌｙｍｐｈｏｃｙｔｅｈｏｍｅｏｓｔａｓｉｓ…     

维生素E琥珀酸酯联合三苯氧胺对乳腺癌荷瘤裸鼠的治疗作用

目的:检测维生素E琥珀酸酯(VES)联合三苯氧胺(TAM)对荷乳腺癌裸鼠的体内肿瘤的抑制作用,并探讨其可能的作用机制.方法:40只裸鼠皮下接种MCF-7乳腺癌细胞建立荷瘤裸鼠模型,分为空白对照组(不予治疗)、TAM治疗组(予5 mg/kg TAM治疗)、VES治疗组(予150 mg/kg VES治疗)、联合治疗组(5 mg/kg TAM 150 mg/kg VES),每组10只,治疗4周后测量各组肿瘤大小,流式细胞仪检测细胞周期变化和细胞表面Fas和FasL表达,免疫组化法检测肿瘤组织Fas和FasL表达,TUNEL法检测细胞凋亡指数的变化.结果:VES联合TAM对荷乳腺癌裸鼠体内肿瘤具有明显的增殖抑制作用,较单用VES或TAM抑制作用增强(P<0.05).细胞周期分析显示,联合药物治疗后肿瘤细胞表现为明显的G0/G1期阻滞,肿瘤组织和细胞表面的Fas和FasL表达上调,同时细胞的凋亡率升高.结论:VES联合TAM对荷乳腺癌裸鼠体内肿瘤具有协同抑制作用,其机制可能与上调肿瘤细胞Fas/FasL表达,促进细胞凋亡有关.     

Effects of Shenmai and Aminophylline on Apoptosis of Small Airway Smooth Muscle Cells and the Expression of Relevant Genes in Rats with Emphysema

In the pathogenesis of emphysema,there arealways accompanied by terminal bronchiole pro-longed abnormal dilation of air chamber,destruc-tion of alveolar wall and small airway wall[1] .Byinvestigating the effects of Shenmai injection( SMI) and aminophylline on apoptosis of small air-way smooth muscle cells ( SASMC) and the Fas/Fas L expression in rat emphysema model inducedby instilling papain into intratracheal,we studytheir protection action and explore their mecha-nism.1　 MATERIAL S…     

柚皮苷通过Fas膜受体通路调控髓核细胞凋亡的机制研究

[目的]观察中药骨碎补单体柚皮苷（Naringin）对人退变椎间盘髓核细胞凋亡的影响,为治疗椎间盘退变提供理论依据。[方法]对体外培养的人退变椎间盘髓核细胞利用番红O染色和甲苯胺蓝染色进行鉴定,并对P3代细胞分别用柚皮苷0 g/mL （对照组）、柚皮苷20 g/mL、柚皮苷40 g/mL、柚皮苷80 g/mL的培养基培养人髓核细胞48 h,噻唑蓝（MTT）法选择其抑制人退变椎间盘髓核细胞凋亡的最佳浓度,并用流式细胞仪测定各组细胞凋亡率。实时定量聚合酶链锁反应（qRT-PCR）检测Fas、FasL、Caspase-8基因表达。蛋白免疫印迹法（Western-blot）检测Fas蛋白表达。[结果]MTT法和流式细胞检测均测定20 g/mL柚皮苷为抑制髓核细胞凋亡的最佳浓度（P<0.05）。qRT-PCR检测20 g/mL柚皮苷能够抑制Fas、FasL、Caspase-8 mRNA表达（P<0.05）。Western-blot检测显示20 g/mL柚皮苷能够抑制Fas、FasL、Caspase-8蛋白表达。[结论]柚皮苷可能通过调控Fas/Fasl死亡受体凋亡通路抑制人退变椎间盘髓核细胞凋亡。     

移植肾AR时病理诊断与细胞凋亡和Fas/FasL的表达

目的:探讨移植肾急性排斥(AR)时细胞凋亡与Fas和Fas配体(FasL)表达的作用及其临床意义。方法:分别用原位末端标记技术(TUNEL法)和免疫组织化学方法检测26例移植肾AR标本中细胞凋亡和Fas/FasL表达情况。结果:细胞凋亡和Fas/FasL表达主要在AR移植肾小管上皮发生,且凋亡指数和Fas/FasL表达与肾组织病理损伤程度平行,与正常肾对照组和移植肾功能稳定组比较差异显著。结论:肾小管上皮细胞凋亡在AR所致的移植肾损伤中起重要作用,Fas/FasL系统可能参与移植肾AR,是造成肾小管上皮细胞凋亡的重要因素。TUNEL法检测细胞凋亡可作为判断移植肾病理变化和预后的重要指标。     

氟美松对急性肝损伤大鼠肝细胞Fas/Fas配体表达和细胞凋亡的影响

目的 :观察内毒素急性肝损伤后肝细胞凋亡和Fas/Fas配体 (FasL)表达的变化 ,以及氟美松对其影响 ,探讨急性肝损伤早期作用机制 .方法 :采用免疫组化和原位杂交技术检测各时相点肝细胞Fas/FasL蛋白和mRNA的表达 ;应用原位末端标记技术对各时相点肝细胞凋亡进行检测 .结果 :内毒素急性肝损伤各时相点大鼠肝细胞Fas/FasL蛋白和mR NA的表达较对照组明显上调 (P <0 .0 5 ) ,且与肝细胞凋亡的增加相一致 ;氟美松可明显减轻炎症反应 ,抑制脂多糖 (LPS)诱导的肝细胞凋亡 ,并使肝细胞Fas/FasL蛋白和mRNA的表达明显下调 (P <0 .0 5 ) .结论 :肝细胞凋亡和Fas/FasL系统活化可能参与内毒素急性肝损伤早期的发病机制 ,而且加重早期肝损伤 ;氟美松可抑制Fas/FasL系统活化 ,阻断和调控凋亡 ,从而减轻肝组织损伤     

三氧化二砷启动人食管癌细胞凋亡机制研究

目的探讨三氧化二砷诱导白血病及实体瘤细胞凋亡的机制。方法以三氧化二砷敏感和不敏感人食管癌细胞株为模型,运用RT-PCR和流式细胞术检测三氧化二砷处理的这两类细胞Fas／FasL表达的改变;同时利用荧光探针检测线粒体功能变化。结果两类细胞有同等水平Fas表达,但均未检测出FasL;三氧化二砷既不能上调Fas也不能诱导FasL表达,但破坏线粒体跨膜电势和氧化-还原系统。结论三氧化二砷诱导人食管癌细胞凋亡不依赖Fas／FasL的启动,而直接作用于线粒体,活化Caspase-3导致凋亡。     

Fas及FasL在胃癌组织非典型增生组织和正常胃粘膜中表达研究

目的:探讨细胞凋亡相关蛋白Fas、FasL在胃癌发生、发展中的作用。方法:应用免疫组化(S-P法)检测Fas、FasL蛋白在胃癌组织、癌旁非典型增生组织中的表达,并应用缺口原位末端标记(TUNEL染色)检测上述组织中的细胞凋亡指数(AI)。结果:Fas、FasL在胃癌及非典型增生组织中的表达显著高于正常胃粘膜(P<0.01),正常胃粘膜组织的凋亡指数显著低于非典型增生组织(P<0.01)和胃癌组织(P<0.05)。结论:Fas、FasL的异常表达可能是胃粘膜癌变过程中细胞凋亡抑制的重要机制之一,并与胃癌的免疫逃逸有关。     

几种ACEI对实验性糖尿病大鼠心肌细胞凋亡和Caspase-3及Fas、FasL基因表达影响的类效应

目的:探讨几种血管紧张素转换酶抑制剂(ACEI)卡托普利、培哚普利、咪达普利、贝那普利、福辛普利对糖尿病大鼠左室心功能及心肌细胞凋亡和凋亡相关蛋白影响的类效应。方法:将48只SD大鼠腹腔注射链脲佐菌素(STZ)诱发糖尿病模型,随机分为5个给药组和1个糖尿病组(每组8只),另选8只做正常对照组。给药组每日分别灌胃给予卡托普利(50 mg/kg),培哚普利(雅施达4 mg/kg),咪达普利(达爽10 mg/kg),贝那普利(洛汀新10 mg/kg),福辛普利(蒙诺10 mg/kg),糖尿病组给予生理盐水灌胃,共8周。对照观察心肌细胞凋亡、心肌组织Fas、FasL蛋白、Caspase-3及其Fas mRNA表达的变化。结果:与正常对照组相比,糖尿病给药组及未给药组的心肌细胞凋亡指数明显增高(均为P<0.05);Caspase-3阳性蛋白表达率明显增高(均为P<0.05);Fas蛋白阳性染色指数增高(均为P<0.05);Fas的mRNA表达明显增高(均为P<0.05)。但糖尿病给药组的较未给药组低(均为P<0.05)。所有各组的FasL蛋白阳性染色指数差异不明显(均为P>0.05)。各种ACEI治疗组之间差异无显著性(均为P>0.05)。结论:糖尿病心肌病的发生、发展过程中有心肌细胞凋亡的变化和Caspase家族及Fas、FasL系统的参与;糖尿病大鼠心肌细胞凋亡指数、心肌组织Fas蛋白以及Fas的mRNA表达水平、Caspase-3蛋白表达水平增高。ACEI干预糖尿病心肌病,能够降低细胞凋亡及Fas基因的表达及Caspase-3蛋白表达水平,且几种ACEI具有类效应。