Department of Ophthalmology, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, No.164 Lanxi Road, Shanghai, 200062, P.R. China.
Shanghai Putuo Central School of Clinical Medicine, Anhui Medical University, Hefei, 230032, China.
BMC Complement Med Ther. 2023 Aug 29;23(1):302. doi: 10.1186/s12906-023-04138-x.
Qijing Mingmu decoction (QJMM), a compound Chinese medicine preparation, which consists of Lycium barbarum, Polygonatum, Ophiopogon japonicus, Poria cocos, Glycyrrhiza, Eclipta prostrata and Ligusticum striatum, has been confirmed to be effective for the treatment of conjunctivochalasis (CCH) in clinic and reduce cellular senescence. However, the underlying mechanism is still unknown. Our previous study revealed that p38-mediated cellular senescence contributed to the pathogenesis of CCH.
To explore whether p38 might be the potential therapeutic target of QJMM for CCH, CCH fibroblasts were treated with QJMM granule and then the effect of QJMM granule on the expression and promoter activity of p38α was determined by western blot and dual luciferase reporter gene assay, respectively. Meanwhile, the influence of QJMM granule on cell proliferation, oxidative stress, cellular senescence and the expression of the cellular senescence-associated genes were measured by corresponding methods.
QJMM granule significantly decreased the protein expression of p38α and p-p38α in CCH fibroblasts in a dose-dependent manner and inhibited p38α promoter activity. QJMM granule as well as the p38 inhibitor SB203580 reduced the level of reactive oxygen species and increased the activity of superoxide dismutase in CCH fibroblasts. QJMM granule and SB203580 promoted cell proliferation and reduced the percentage of SA-β-Gal-positive cells. The mRNA and protein expression of p53 and p21 was remarkably down-regulated by QJMM granule as well as SB203580 and that of SMP30 was up-regulated in CCH fibroblasts.
Our findings demonstrated that QJMM granule was effective for alleviating cellular senescence of CCH fibroblasts by p38 MAPK signaling and the followed p53/p21 signaling.
杞菊明目汤(QJMM)是一种复方中药制剂,由枸杞、黄精、麦冬、茯苓、甘草、旱莲草和川芎组成,已被临床证实可有效治疗结膜松弛症(CCH),并可减轻细胞衰老。然而,其作用机制尚不清楚。我们之前的研究表明,p38 介导的细胞衰老导致了 CCH 的发病机制。
为了探讨 p38 是否可能是 QJMM 治疗 CCH 的潜在治疗靶点,用 QJMM 颗粒处理 CCH 成纤维细胞,然后通过 Western blot 和双荧光素酶报告基因检测分别测定 QJMM 颗粒对 p38α 的表达和启动子活性的影响。同时,通过相应的方法测定 QJMM 颗粒对细胞增殖、氧化应激、细胞衰老和细胞衰老相关基因表达的影响。
QJMM 颗粒以剂量依赖的方式显著降低 CCH 成纤维细胞中 p38α 和 p-p38α 的蛋白表达,并抑制 p38α 启动子活性。QJMM 颗粒和 p38 抑制剂 SB203580 降低了 CCH 成纤维细胞中活性氧的水平,增加了超氧化物歧化酶的活性。QJMM 颗粒和 SB203580 促进细胞增殖,降低了 SA-β-Gal 阳性细胞的比例。QJMM 颗粒和 SB203580 显著下调了 CCH 成纤维细胞中 p53 和 p21 的 mRNA 和蛋白表达,上调了 SMP30 的表达。
我们的研究结果表明,QJMM 颗粒通过 p38 MAPK 信号通路及随后的 p53/p21 信号通路,对 CCH 成纤维细胞的衰老具有缓解作用。