Antibiotics induce apoptosis of human peritoneal mesothelial cells
Antibiotics induce apoptosis of human peritoneal mesothelial cells
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DOI：
10.1046/j.1440-1797.2003.00149.x
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发表时间：
2003-06
期刊：
影响因子：
2.5
通讯作者：
C. Fang;C. Yen;T. Tsai;Ruey-Hwa Chen;Po-Huang Lee;Y. Tomino
中科院分区：
文献类型：
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作者：
C. Fang;C. Yen;T. Tsai;Ruey-Hwa Chen;Po-Huang Lee;Y. Tomino
腹膜间皮细胞是腹膜的重要组成部分。腹膜内使用几种抗生素治疗细菌性腹膜炎是目前的临床实践。我们的前期研究表明头孢噻吩（CPL）和头孢噻肟（CFT）对人腹膜间皮细胞（HPMC）具有细胞毒性作用，但其确切的细胞毒性机制尚未阐明。本研究采用流式细胞术、原位末端标记（TUNEL）法和电镜技术检测HPMCs凋亡情况。免疫荧光染色用于评价细胞色素c分布模式。蛋白质印迹法用于评估凋亡信号蛋白。CPL（0.5 mg/mL）和CFT（1 mg/mL）可诱导HPMCs凋亡，而头孢唑啉（0.5 mg/mL）和头孢曲松（0.5 mg/mL）不能诱导HPMCs凋亡。虽然CFT或CPL处理的细胞的DNA含量降低，但通过流式细胞术测定，头孢唑啉和头孢曲松没有这种作用。CFT或CPL处理的细胞在电镜下和TUNEL染色中都显示出凋亡的特征。然而，头孢唑啉和头孢曲松产生与培养基对照相同的结果。CFT和CPL还可增加Bax和p53的表达，并使细胞色素c从线粒体向胞浆移位。经CFT处理而非经CPL处理的HPMC诱导半胱天冬酶原-3裂解以形成活性半胱天冬酶-3。结论：头孢噻肟和头孢噻吩在体外诱导人乳头状肌细胞凋亡。信号转导可能通过线粒体途径。
SUMMARY: The peritoneal mesothelial cell is a critical component of the peritoneal membrane. The intraperitoneal use of several antibiotics to treat bacterial peritonitis is current clinical practice. Our previous study showed that cephalothin (CPL) and cefotaxime (CFT) have cytotoxic effects on human peritoneal mesothelial cells (HPMC), however, the exact mechanism of cytotoxicity has not been elucidated. In the present study, flow cytometry, TdT‐mediated dUTP nick‐end labelling (TUNEL) staining and electron microscopy were used to detect the apoptosis of HPMCs. Immunofluorescent staining was used to evaluate the cytochrome c distribution pattern. Western blotting was used to assess apoptotic signalling proteins. We found that CPL (0.5 mg/mL) and CFT (1 mg/mL) induced apoptosis of HPMCs, whereas cefazolin (0.5 mg/mL) and ceftriaxone (0.5 mg/mL) failed to induce apoptosis of HPMCs. While the DNA content of CFT‐ or CPL‐treated cells was reduced, as determined by flow cytometry, cefazolin and ceftriaxone had no such effect. The CFT‐ or CPL‐treated cells displayed the features of apoptosis both under the electron microscope and by using TUNEL staining. However, cefazolin and ceftriaxone produced the same result as the medium controls. Furthermore, CFT and CPL increased the expression of Bax and p53, and caused the translocation of cytochrome c from the mitochondria to the cytoplasm. The HPMC treated by CFT but not by CPL induced the cleavage of procaspase‐3 to form active caspase‐3. In conclusion, cefotaxime and cephalothin induce apoptosis of HPMCs in vitro. Signal transduction may be through the mitochondrial pathway.