国际眼科杂志
國際眼科雜誌
국제안과잡지
INTERNATIONAL JOURNAL OF OPHTHALMOLOGY
2014年
2期
214-218
,共5页
罗莎莎%范莲%孙松%武志峰
囉莎莎%範蓮%孫鬆%武誌峰
라사사%범련%손송%무지봉
羟基喜树碱%丝裂霉素C%人眼Tenon's囊成纤维细胞%抑制作用
羥基喜樹堿%絲裂黴素C%人眼Tenon's囊成纖維細胞%抑製作用
간기희수감%사렬매소C%인안Tenon's낭성섬유세포%억제작용
hydroxycamptothecin%mitomycin C%human Tenon's capsule fibroblasts%inhibition
目的：研究羟基喜树碱（ HCPT ）对人眼Tenon's囊成纤维细胞（ human Tenon's capsule fibroblasts ， HTFs）的细胞周期、细胞毒性的影响，探讨其抗纤维化作用机制。<br> 方法：取本院眼库新鲜眼球（＜6 h ）的Tenon's囊组织，用组织块培养法进行成纤维细胞体外培养；流式细胞仪测定HCPT和丝裂霉素C（MMC）对HTFs细胞周期的影响；台盼蓝染色法鉴定HCPT和MMC对HTFs的抑制作用是否由药物的细胞毒性引起；RT-PCR检测HCPT、MMC作用24 h后HTFs的Smad7 mRNA基因表达。<br> 结果：HTFs体外生长良好，可用于实验研究。不同浓度HCPT （0，0．25，1，4mg/L）作用后的HTFs细胞周期与对照组相比，G2期细胞百分数的组间差异均有统计学意义（P＜0．05）；不同浓度MMC（0，0．025，0．1，0．4mg/L）作用后的HTFs细胞周期与对照组相比，G1期细胞百分数的组间差异均有统计学意义（ P＜0．05）。不同浓度HCPT和MMC作用后的HTFs活细胞率和空白对照组比较，差异均无统计学意义（P＞0．05）。4mg/L HCPT，0．4mg/L MMC作用HTFs 24 h后，Smad7 mRNA表达水平与空白对照组比较差异有统计学意义（P＜0．05）。<br> 结论：HCPT将HTFs阻滞于G2期，MMC将HTFs阻滞于G1期；HCPT，MMC抑制HTFs的作用和药物的细胞毒性无关；HCPT抑制 HTFs 的增殖可能是通过上调 Smad7 mRNA的表达阻断TGF-β信号通路实现的。
目的：研究羥基喜樹堿（ HCPT ）對人眼Tenon's囊成纖維細胞（ human Tenon's capsule fibroblasts ， HTFs）的細胞週期、細胞毒性的影響，探討其抗纖維化作用機製。<br> 方法：取本院眼庫新鮮眼毬（＜6 h ）的Tenon's囊組織，用組織塊培養法進行成纖維細胞體外培養；流式細胞儀測定HCPT和絲裂黴素C（MMC）對HTFs細胞週期的影響；檯盼藍染色法鑒定HCPT和MMC對HTFs的抑製作用是否由藥物的細胞毒性引起；RT-PCR檢測HCPT、MMC作用24 h後HTFs的Smad7 mRNA基因錶達。<br> 結果：HTFs體外生長良好，可用于實驗研究。不同濃度HCPT （0，0．25，1，4mg/L）作用後的HTFs細胞週期與對照組相比，G2期細胞百分數的組間差異均有統計學意義（P＜0．05）；不同濃度MMC（0，0．025，0．1，0．4mg/L）作用後的HTFs細胞週期與對照組相比，G1期細胞百分數的組間差異均有統計學意義（ P＜0．05）。不同濃度HCPT和MMC作用後的HTFs活細胞率和空白對照組比較，差異均無統計學意義（P＞0．05）。4mg/L HCPT，0．4mg/L MMC作用HTFs 24 h後，Smad7 mRNA錶達水平與空白對照組比較差異有統計學意義（P＜0．05）。<br> 結論：HCPT將HTFs阻滯于G2期，MMC將HTFs阻滯于G1期；HCPT，MMC抑製HTFs的作用和藥物的細胞毒性無關；HCPT抑製 HTFs 的增殖可能是通過上調 Smad7 mRNA的錶達阻斷TGF-β信號通路實現的。
목적：연구간기희수감（ HCPT ）대인안Tenon's낭성섬유세포（ human Tenon's capsule fibroblasts ， HTFs）적세포주기、세포독성적영향，탐토기항섬유화작용궤제。<br> 방법：취본원안고신선안구（＜6 h ）적Tenon's낭조직，용조직괴배양법진행성섬유세포체외배양；류식세포의측정HCPT화사렬매소C（MMC）대HTFs세포주기적영향；태반람염색법감정HCPT화MMC대HTFs적억제작용시부유약물적세포독성인기；RT-PCR검측HCPT、MMC작용24 h후HTFs적Smad7 mRNA기인표체。<br> 결과：HTFs체외생장량호，가용우실험연구。불동농도HCPT （0，0．25，1，4mg/L）작용후적HTFs세포주기여대조조상비，G2기세포백분수적조간차이균유통계학의의（P＜0．05）；불동농도MMC（0，0．025，0．1，0．4mg/L）작용후적HTFs세포주기여대조조상비，G1기세포백분수적조간차이균유통계학의의（ P＜0．05）。불동농도HCPT화MMC작용후적HTFs활세포솔화공백대조조비교，차이균무통계학의의（P＞0．05）。4mg/L HCPT，0．4mg/L MMC작용HTFs 24 h후，Smad7 mRNA표체수평여공백대조조비교차이유통계학의의（P＜0．05）。<br> 결론：HCPT장HTFs조체우G2기，MMC장HTFs조체우G1기；HCPT，MMC억제HTFs적작용화약물적세포독성무관；HCPT억제 HTFs 적증식가능시통과상조 Smad7 mRNA적표체조단TGF-β신호통로실현적。
AIM:To investigate the effect of different concentrations of 10-hydroxycamptothecin ( HCPT) on the cell cycle and cytotoxicity of human Tenon's capsule fibroblasts ( HTFs) , and to explore its mechanism in anti-fibrosis.METHODS:The Tenon's capsule tissue of fresh eyes (<6h ) of our hospital eye bank was taken, in vitro culture of fibroblasts was done by tissue block culture;A flow cytometry ( FCM) was used to evaluate the effect of different concentrations of HCPT (0, 0.25, 1, 4mg/L) and mitomycin C ( MMC: 0, 0.025, 0.1, 0.4mg/L) on HTFs cell cycle; Trypan blue staining was adopted to determine whether the inhibitory effect of HCPT and MMC on HTFs was caused by their cytotoxicity;RT-PCR was employed to detect the level of Smad7mRNA gene expression of HTFs after HCPT and MMC were used for 24h. <br> RESULTS:HTFs were cultured successfully in vitro, and can be used in our study; compared with the control groups, the cell cycle of HTFs which was affected by different concentrations of HCPT was significantly different in G2 phase (P<0.05).As for MMC, there were significant differences in G1 phase between groups with different concentrations of MMC and the control group (P<0.05).There was no significant difference in the rate of HTFs living cells between HCPT group ( or MMC group) to which HCPT and MMC were added for 24h and the control group (P>0.05).After HFTs was affected by 0.4mg/L MMC or 4mg/L HCPT for 24h, RT PCR found that the level of Smad7 mRNA expression was significantly increased (P<0.05). <br> CONCLUSION:HCPT mainly blocks HTFs in G2 phase, MMC mainly impacts the G1 phase.The inhibitory effect of HCPT and MMC on HTFs proliferation is not relevant to cytotoxicity induced by the two drugs.The mechanism that HCPT inhibits the proliferation of HTFs may be due to the increasing Smad7 mRNA expression to block TGF-βsignaling pathway.
