摘要
目的 探讨热休克蛋白 90 (HSP90 )与糖皮质激素受体 (GR)比例失调对T淋巴细胞凋亡的作用 ,同时观察地塞米松 (Dex)对HSP90和GRmRNA表达的影响。方法 选择 10例哮喘患者和 7例正常对照 ,利用格尔得霉素 (GA)特异性阻断HSP90的作用 ,造成功能性HSP90与GR比例降低 ,用碘化丙啶 (PI)染色流式细胞仪计数凋亡细胞的方法 ,观察用Dex和GA处理的T细胞凋亡情况 ;用逆转录聚合酶链反应 (RT PCR)的方法观察Dex和GA处理对哮喘T淋巴细胞HSP90和GRmRNA表达的影响。结果 Dex可明显诱导哮喘T淋巴细胞的凋亡 (33 8%± 3 2 %比 2 3 2 %± 1 5 % ,P <0 0 1) ,GA对T淋巴细胞的凋亡无明显影响 ,却可阻断Dex诱导T淋巴细胞凋亡的作用 (2 4 5 %±6 0 %比 33 8%± 3 2 % ,P <0 0 1) ;Dex对正常人T淋巴细胞的凋亡有诱导作用 ,但其程度较弱(2 5 9%± 3 5 %比 2 3 1%± 1 5 % ,P <0 0 5 )。Dex可抑制哮喘患者T淋巴细胞HSP90和GR的表达(分别为 1 2 3± 0 16比 1 6 8± 0 38和 0 42± 0 0 6比 0 5 4± 0 0 7,P均 <0 0 5 ) ,GA对Dex的这一作用有阻断作用 ,但GA本身对HSP90和GRmRNA的表达无明显影响。结论 HSP90 /GR比例降低可影响Dex诱导的T细胞凋亡 ;Dex对哮喘患者GR和HSP90mRNA的表达有下调作用 。
Objective Using GA(geldanamycine), which specifically binding to HSP90, to induce the imbalance of HSP90/GR in function(low ratio)and analyzing the effect of low HSP90/GR ratio on T lymphocytes apoptosis induced by Dex from asthmatic patients. The expression of HSP90 and GR mRNA of T lymphocytes influenced by Dex and GA was also studied. Methods Peripheral blood T lymphocytes were enriched from 10 asthmatic subjects and 7 healthy volunteers by nylon column. T lymphocytes were cultured in vitro with Dex and /or GA for 72 hours. Apoptosis of T lymphocytes were measured by propidium iodide staining and flowcytometry. With reverse transcription polymerase chain reaction(RT PCR),the expression of HSP90 and GR mRNA was detected. Results Dex could obviously induce the apoptosis of T lymphocytes from asthmatic patients(33 8%±3 2% vs 23 2%±1 5% , P <0 01). GA had no effect on the apoptosis of T lymphocytes but could inhibit the effect of Dex(24 5%±6 0% vs 33 8%±3 2%, P <0 01).Dex also had the effect of inducing apoptosis of T lymphocytes from health volunteers but the effect was less potent than that from asthmatic patients(25 9%±3 5% vs 23 1%±1 5%, P <0 05). Dex inhibited the expression of HSP90 and GR mRNA of T lymphocytes from asthmatic patients(1 23±0 16 vs 1 68±0 38 and 0 42±0 06 vs 0 54±0 07,respectively, P all<0 05). GA could interrupt the inhibiting effect of Dex on the expression of HSP90 and GR mRNA but had no effect on it. Conclusion The low ratio of HSP90/GR could reduce the inducing apoptosis effect of Dex. Dex could down regulate the mRNA expression of HSP90 and GR and GA could interrupt the inhibiting effect of Dex.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2001年第24期1496-1499,共4页
National Medical Journal of China
