目的探讨表面活性蛋白A(surfactant protein A,SP-A)不同亚型(SP-A1,SP-A2)对脂多糖(lipopolysaccharide,LPS)诱导的人近曲肾小管上皮细胞(HK-2)IL-8表达的影响。方法体外培养HK-2细胞,分别转染SP-A1、SP-A2及阴性对照siRNA,实时定量PC...目的探讨表面活性蛋白A(surfactant protein A,SP-A)不同亚型(SP-A1,SP-A2)对脂多糖(lipopolysaccharide,LPS)诱导的人近曲肾小管上皮细胞(HK-2)IL-8表达的影响。方法体外培养HK-2细胞,分别转染SP-A1、SP-A2及阴性对照siRNA,实时定量PCR检测SP-A1及SP-A2 mRNA表达水平,并给予1μg/ml LPS刺激8h,ELISA观察转染不同siRNA后IL-8蛋白表达变化。结果转染SP-A1、SP-A2 siRNA可分别显著下调转染SP-A1、SP-A2的mRNA表达水平(P<0.05);1μg/ml LPS刺激8h可引起HK-2细胞IL-8蛋白表达显著增加(P<0.05),低表达SP-A2可下调LPS诱导的IL-8表达(P<0.05)。结论 LPS刺激下肾小管上皮细胞IL-8表达增加与SP-A2的表达有关,SP-A2可能在肾脏炎性反应中发挥重要作用。展开更多
目的:目的研究167例乙肝病毒截短型表面抗原中蛋白(C-terminally truncated middle size surface proteins,MHBst167)和X蛋白(hepatitis B virus X protein,HBx)对肾小管上皮细胞核转录因子κB(nuclear factor-kappaB,NF-κB)活化的影...目的:目的研究167例乙肝病毒截短型表面抗原中蛋白(C-terminally truncated middle size surface proteins,MHBst167)和X蛋白(hepatitis B virus X protein,HBx)对肾小管上皮细胞核转录因子κB(nuclear factor-kappaB,NF-κB)活化的影响及其相关机制探讨。方法:肾小管上皮细胞系(HK-2)转染mhbst167或(和)hbx后,蛋白印迹法检测NF-κB核易位及其抑制蛋白(inhibitor of nuclear factor-kappa B,IκBα)磷酸化水平的变化,凝胶电泳迁移率和双萤光素酶报告基因分析进一步检测NF-κB活性;通过对蛋白激酶C(protein kinase C,PKC)活性和细胞外调节激酶(extracellular regulated protein kinases,ERK)磷酸化水平检测探讨NF-κB活化的机制。结果:HK-2细胞转染mhbst167或(和)hbx基因后,NF-κB核易位、磷酸化IκBα、κB结合活性及κB基因转录均增加(P<0.05);且PKC激酶活性和磷酸化ERK水平也增加(P<0.05),而Raf蛋白均无表达。结论:在肾小管上皮细胞中,MHBst167/HBx可能通过PKC/ERK通路(非Raf依赖性)活化NF-κB。展开更多
BACKGROUND: Ocular trauma or disease may lead to severe corneal opacification and, consequently, severe loss of vision as a result of complete loss of corneal epithelial stem cells. Transplantation of autologous corne...BACKGROUND: Ocular trauma or disease may lead to severe corneal opacification and, consequently, severe loss of vision as a result of complete loss of corneal epithelial stem cells. Transplantation of autologous corneal stem-cell sources is an alternative to allograft transplantation and does not require immunosuppr ession, but it is not possible in many cases in which bilateral disease produces total corneal stem-cell deficiency in both eyes. We studied the use of autolog ous oral mucosal epithelial cells as a source of cells for the reconstruction of the corneal surface. METHODS: We harvested 3-by-3-mm specimens of oral mucos al tissue from four patients with bilateral total corneal stem-cell deficiencie s. Tissue-engineered epithelial-cell sheets were fabricated exvivo by culturin g harvested cells for two weeks on temperature responsive cell-culture surfaces with 3T3 feeder cells that had been treated with mitomycin C. After conjunctiva l fibrovascular tissue had been surgically removed from the ocular surface, shee ts of cultured autologous cells that had been harvested with a simple reducedtem perature treatment were transplanted directly to the denuded corneal surfaces (o ne eye of each patient) without sutures. RESULTS: Complete reepithelialization o f the corneal surfaces occurred within one week in all four treated eyes. Cornea l transparency was restored and postoperative visual acuity improved remarkably in all four eyes. During a mean follow-up period of 14 months, all corneal surf aces remained transparent. There were no complications. CONCLUSIONS: Sutureless transplantation of carrier-free cell sheets composed of autologous oral mucosal epithelial cells may be used to reconstruct corneal surfaces and can restore vi sion in patients with bilateral severe disorders of the ocular surface.展开更多
文摘目的:目的研究167例乙肝病毒截短型表面抗原中蛋白(C-terminally truncated middle size surface proteins,MHBst167)和X蛋白(hepatitis B virus X protein,HBx)对肾小管上皮细胞核转录因子κB(nuclear factor-kappaB,NF-κB)活化的影响及其相关机制探讨。方法:肾小管上皮细胞系(HK-2)转染mhbst167或(和)hbx后,蛋白印迹法检测NF-κB核易位及其抑制蛋白(inhibitor of nuclear factor-kappa B,IκBα)磷酸化水平的变化,凝胶电泳迁移率和双萤光素酶报告基因分析进一步检测NF-κB活性;通过对蛋白激酶C(protein kinase C,PKC)活性和细胞外调节激酶(extracellular regulated protein kinases,ERK)磷酸化水平检测探讨NF-κB活化的机制。结果:HK-2细胞转染mhbst167或(和)hbx基因后,NF-κB核易位、磷酸化IκBα、κB结合活性及κB基因转录均增加(P<0.05);且PKC激酶活性和磷酸化ERK水平也增加(P<0.05),而Raf蛋白均无表达。结论:在肾小管上皮细胞中,MHBst167/HBx可能通过PKC/ERK通路(非Raf依赖性)活化NF-κB。
文摘BACKGROUND: Ocular trauma or disease may lead to severe corneal opacification and, consequently, severe loss of vision as a result of complete loss of corneal epithelial stem cells. Transplantation of autologous corneal stem-cell sources is an alternative to allograft transplantation and does not require immunosuppr ession, but it is not possible in many cases in which bilateral disease produces total corneal stem-cell deficiency in both eyes. We studied the use of autolog ous oral mucosal epithelial cells as a source of cells for the reconstruction of the corneal surface. METHODS: We harvested 3-by-3-mm specimens of oral mucos al tissue from four patients with bilateral total corneal stem-cell deficiencie s. Tissue-engineered epithelial-cell sheets were fabricated exvivo by culturin g harvested cells for two weeks on temperature responsive cell-culture surfaces with 3T3 feeder cells that had been treated with mitomycin C. After conjunctiva l fibrovascular tissue had been surgically removed from the ocular surface, shee ts of cultured autologous cells that had been harvested with a simple reducedtem perature treatment were transplanted directly to the denuded corneal surfaces (o ne eye of each patient) without sutures. RESULTS: Complete reepithelialization o f the corneal surfaces occurred within one week in all four treated eyes. Cornea l transparency was restored and postoperative visual acuity improved remarkably in all four eyes. During a mean follow-up period of 14 months, all corneal surf aces remained transparent. There were no complications. CONCLUSIONS: Sutureless transplantation of carrier-free cell sheets composed of autologous oral mucosal epithelial cells may be used to reconstruct corneal surfaces and can restore vi sion in patients with bilateral severe disorders of the ocular surface.
