Li Na, Deng Xin-Guo, Zhang Shi-Hua, He Mei-Feng, Zhao Dong-Qing
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, Guangdong Province, China ; Department of Ophthalmology, Huizhou First Hospital, Huizhou 516001, Guangdong Province, China.
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, Guangdong Province, China.
Int J Ophthalmol. 2014 Dec 18;7(6):947-51. doi: 10.3980/j.issn.2222-3959.2014.06.06. eCollection 2014.
To discuss the effects of different concentrations of tetramethylpyrazine (TMP), an active constituent of Chinese herb, on damaged Shandong human corneal epithelial cell (SDHCEC) induced by hydrogen peroxide.
We detected the combined effects of TMP with concentrations ranging from 4 mg/mL to 0.03 mg/mL and 800 µM hydrogen peroxide on SDHCEC. The methyl thiazolyl tetrazolium (MTT) assay was processed at 3, 6 and 12h separately while the detection of cell apoptosis at 6h only by flow cytometry.
The viability of SDHCEC with 0.5 mg/mL, 0.25 mg/mL, 0.125 mg/mL and 0.06 mg/mL TMP joint with 800 µM hydrogen peroxide at 3h and 6h was significantly higher than that with 800 µM hydrogen peroxide only, P<0.05. However, except 0.25 mg/mL, TMP with other concentrations joint with 800 µM hydrogen peroxide at 12h could not significantly inhibit decreased SDHCEC viability induced by 800 µM hydrogen peroxide. At 12h, TMP of 0.5 mg/mL, 0.25 mg/mL, 0.125 mg/mL and 0.06 mg/mL could significantly inhibit SDHCEC early apoptosis induced by 800 µM hydrogen peroxide, most remarkable at 0.25 mg/mL TMP, P<0.05.
Our results suggested that hydrogen peroxide can induce apoptosis related damage to SDHCEC. TMP can protect SDHCEC from the damage, and the protective effects may be associated with its anti-apoptosis mechanism.
探讨中药有效成分川芎嗪(TMP)不同浓度对过氧化氢诱导损伤的山东人角膜上皮细胞(SDHCEC)的影响。
检测浓度范围为4mg/mL至0.03mg/mL的TMP与800μM过氧化氢对SDHCEC的联合作用。分别在3、6和12小时进行甲基噻唑基四氮唑(MTT)检测,仅在6小时通过流式细胞术检测细胞凋亡。
在3小时和6小时,0.5mg/mL、0.25mg/mL、0.125mg/mL和0.06mg/mL的TMP与800μM过氧化氢联合作用下的SDHCEC活力显著高于仅用800μM过氧化氢处理的细胞,P<0.05。然而,在12小时时,除0.25mg/mL外,其他浓度的TMP与800μM过氧化氢联合不能显著抑制800μM过氧化氢诱导的SDHCEC活力下降。在12小时时,0.5mg/mL、0.25mg/mL、0.125mg/mL和0.06mg/mL的TMP可显著抑制800μM过氧化氢诱导的SDHCEC早期凋亡,其中0.25mg/mL的TMP效果最显著,P<0.05。
我们的结果表明,过氧化氢可诱导SDHCEC发生凋亡相关损伤。TMP可保护SDHCEC免受损伤,其保护作用可能与其抗凋亡机制有关。