Influences of methyl-β-cyclodextrin-caused caveolae destruction on TGF-β/Smad signaling pathway and on proliferation of type II alveolar epithelial cells

Wang Qin; Wang Jianchun; Li Yuying; Wang Guansong

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Influences of methyl-β-cyclodextrin-caused caveolae destruction on TGF-β/Smad signaling pathway and on proliferation of type II alveolar epithelial cells

更新时间：2025-01-15

- Influences of methyl-β-cyclodextrin-caused caveolae destruction on TGF-β/Smad signaling pathway and on proliferation of type II alveolar epithelial cells
- Journal of Army Medical University Vol. 33, Issue 8, Pages: 780-784(2011)
- 作者机构：
  
  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
- 作者简介：
- 基金信息：
- DOI：
  CLC： R322.35;R329.28;R363.13
- Published：2011，
  
  Published Online：30 April 2011，
- 稿件说明：
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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]. Journal of army medical university, 2011, 33(8): 780-784.
DOI：

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]. Journal of army medical university, 2011, 33(8): 780-784. DOI：

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(

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（

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（

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 i

dentified 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)%

(54.83±5.67)%

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)%

(8.36±0.64)%

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)%

(49.20±4.44)%

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.

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 cellscaveolaeCaveolin-1TGF-β/Smad signaling pathway

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