[1]王勤,王建春,李玉英,等.甲基-β-环糊精对肺泡Ⅱ型上皮细胞增殖和TGF-β/Smad信号通路的影响[J].陆军军医大学学报(原第三军医大学学报),2011,33(08):780-784.
 Wang Qin,Wang Jianchun,Li Yuying,et al.Influences of methyl-β-cyclodextrin-caused caveolae destruction on TGF-β/Smad signaling pathway and on proliferation of type II alveolar epithelial cells[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(08):780-784.
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甲基-β-环糊精对肺泡Ⅱ型上皮细胞增殖和TGF-β/Smad信号通路的影响(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第08期
页码:
780-784
栏目:
论著
出版日期:
2011-04-30

文章信息/Info

Title:
Influences of methyl-β-cyclodextrin-caused caveolae destruction on TGF-β/Smad signaling pathway and on proliferation of type II alveolar epithelial cells
作者:
王勤王建春李玉英王关嵩
第三军医大学新桥医院全军呼吸内科研究所, 全军呼吸病研究重点实验室
Author(s):
Wang Qin Wang Jianchun Li Yuying Wang Guansong
Institute of Respiratory Diseases, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China
关键词:
肺泡Ⅱ型上皮细胞小窝小窝蛋白-1转化生长因子β
Keywords:
typeⅡalveolar epithelial cells Caveolae Caveolin-1 TGF-β/Smad signaling pathway
分类号:
R322.35;R329.28;R363.13
文献标志码:
A
摘要:
目的    观察甲基-β-环糊精(methyl-β-cyclodextrin,MβCD)破坏小窝(caveolae)对肺泡Ⅱ型上皮细胞(AECⅡ)增殖和TGF-β/Smad信号通路的影响。    方法    分离培养大鼠AECⅡ,免疫荧光双标检测小窝蛋白-1(caveolin-1)和TGF-β受体Ⅰ(TβR-Ⅰ)表达和分布关系。以MβCD(5 mmol/L)破坏AECⅡ细胞膜Caveolae,设空白对照组,非离子去污剂法提取脂筏检测TβR-Ⅰ与Caveolin-1在细胞膜上的分布;Western blot检测Caveolin-1和磷酸化Smad2(pSmad2)表达;四甲基偶氮唑盐(MTT)法检测细胞增殖能力。    结果    荧光双标和脂筏提取结果提示TβR-Ⅰ主要分布于Caveolae区域,MβCD破坏Caveolae后,TβR-Ⅰ重新分布于细胞膜上非脂筏区域;MβCD干扰组Caveolin-1蛋白表达(24.53±3.24)%较正常对照组(54.83±5.67)%显著下调(P<0.01),而TGF-β/Smad信号通路下游分子pSmad2蛋白表达(10.93±1.11)%较对照组(8.36±0.64)%上调(P<0.05);MβCD干扰组细胞增殖率(31.00±4.18)%较对照组(49.20±4.44)%显著降低(P<0.01)。    结论    MβCD破坏Caveolae结构可抑制AECⅡ增殖,其机制可能与TβR-Ⅰ在非脂筏区域的聚集和Caveolin-1下调导致的TGF-β/Smad信号通路增强有关。
Abstract:
Objective    To study the influences of methyl-β-cyclodextrin (MβCD)-caused caveolae destruction on proliferation of type Ⅱ alveolar epithelial cells (AECs Ⅱ) and on TGF-β/Smad signaling pathway in AECs Ⅱ.     Methods    Rat AECs Ⅱ were isolated through enzyme digestion, and then identified through immunofluorescence assay. The distribution of caveolin-1 (a caveolae-specific protein) and type I TGF-β receptor (TβR-Ⅰ) in AECs Ⅱ cell membranes was analyzed with double-labeling immunofluorescence assay and confocal laser scanning microscopy. AECs Ⅱ were divided into a treatment group and a control group. MβCD (5 mmol/L in DMEM) was added into the treatment group to destroy caveolae of AECs Ⅱ, while DMEM was added into the control group. Lipid rafts were extracted from AECs Ⅱ by nonionic detergent method, and the distribution of caveolin-1 and TβR-Ⅰ in cell membranes of treated AECs Ⅱ was analyzed through SDS-PAGE. The expression of caveolin-1 and phosphorylated Smad2 (pSmad2, a downstream molecule of TGF-β/Smad signaling pathway) in AECs Ⅱ was analyzed through Western blotting. The proliferation rate of AECs Ⅱ was analyzed through methyl thiazolyl tetrazolium method.     Results    The double-labeling immunofluorescence assay and lipid raft extraction showed that TβR-Ⅰ was mainly distributed in caveolae of cell membrane and, after MβCD treatment, was re-distributed in non-raft domains. The expression of caveolin-1 in AECs Ⅱ of the treatment group was significantly lower than that of the control group [(24.53±3.24)% vs (54.83±5.67)%, P<0.01]. The expression of pSmad2 in AECs Ⅱ of the treatment group was significantly higher than that of the control group [(10.93±1.11)% vs (8.36±0.64)%, P<0.05]. The proliferation rate of AECs Ⅱ of the treatment group is significantly lower than that of the control group (31.00±4.18)% vs (49.20±4.44)%, P<0.01).     Conclusion    MβCD-caused caveolae destruction can inhibit the proliferation of AECs Ⅱ, probably through a mechanism of enhancing TGF-β/Smad signaling pathway with TβR-Ⅰ accumulation in non-raft domains and caveolin-1 downregulation.

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更新日期/Last Update: 2011-04-13