Guo Yuanyuan, Ni Shouxiang, Zhou Weiyan, Hou Jiangping, Shen Jiaquan
Department of Ophthalmology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong, China.
J Ophthalmol. 2020 Feb 10;2020:5703286. doi: 10.1155/2020/5703286. eCollection 2020.
To evaluate the role of SPARC in the antiproliferation effect of MMC on human Tenon's fibroblasts (HTF).
Sixteen PACG patients aged 59 ± 10 years (31-72 years), including 6 males and 10 females, were recruited. Tenon tissue was harvested during filtering surgery. Cell density was evaluated after MMC application with different concentrations and application times, by which the optimized MMC application modality was determined. MMC, si-SPARC, or SPARC protein was used when needed to evaluate the cell densities under different conditions, by which the role of SPARC in MMC-mediated antifibrotic process was identified.
Considering that the cell densities, as well as SPARC expression on mRNA and protein levels, are relatively stable when the MMC concentration is higher than 0.02% and exposure time longer than 90 s, we chose the MMC application pattern with 0.02% and 90 s as an optimized pattern for the downstream work. Compared to control, the si-SPARC and MMC downregulated the SPARC protein by 91% ( < 0.01) and 65% ( < 0.01) and 65% ( < 0.01) and 65% ( < 0.01) and 65% ( < 0.01) and 65% ( < 0.01) and 65% ( < 0.01) and 65% ( < 0.01) and 65% (.
This study demonstrates that in HTF, (1) MMC downregulates the expression of SPARC in protein and mRNA levels; (2) SPARC depletion has synergistic effect on the antifibrotic effect of MMC; and (3) reactive oxygen species are the possible mediator in the antifibrotic effect of MMC and si-SPARC.
评估富含半胱氨酸的酸性分泌蛋白(SPARC)在丝裂霉素C(MMC)对人Tenon囊成纤维细胞(HTF)抗增殖作用中的作用。
招募了16例年龄为59±10岁(31 - 72岁)的原发性闭角型青光眼(PACG)患者,其中男性6例,女性10例。在滤过性手术中获取Tenon囊组织。应用不同浓度和作用时间的MMC后评估细胞密度,以此确定MMC的最佳应用方式。必要时使用MMC、SPARC小干扰RNA(si - SPARC)或SPARC蛋白来评估不同条件下的细胞密度,从而确定SPARC在MMC介导的抗纤维化过程中的作用。
考虑到当MMC浓度高于0.02%且暴露时间长于90秒时,细胞密度以及SPARC在mRNA和蛋白水平的表达相对稳定,我们选择0.02%和90秒的MMC应用模式作为下游工作的优化模式。与对照组相比,si - SPARC和MMC使SPARC蛋白表达分别下调91%(P<0.01)和65%(P<0.01)。
本研究表明,在HTF中,(1)MMC在蛋白和mRNA水平下调SPARC的表达;(2)SPARC缺失对MMC的抗纤维化作用具有协同效应;(3)活性氧可能是MMC和si - SPARC抗纤维化作用的介质。