苄基异喹啉生物碱主要通过抑制TGF-β1/Smads和ERK1/2信号通路蛋白来抑制肺成纤维细胞的活化。

Benzylisoquinoline alkaloids inhibit lung fibroblast activation mainly via inhibiting TGF-β1/Smads and ERK1/2 pathway proteins.

作者信息

Ren Hui-Li, Zhang Jia-Hua, Xiao Jun-Hua

机构信息

Department of Pharmacology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

The Key Laboratory for Drug Target Researches and Pharmacodynamic Evaluation of Hubei Province, Wuhan 430030, China.

出版信息

Heliyon. 2023 Jun 1;9(6):e16849. doi: 10.1016/j.heliyon.2023.e16849. eCollection 2023 Jun.

BACKGROUNDS

Liensinine (Lien), Neferine (Nef), Isoliensinine (Iso) and Tetrandrine (Tet), benzylisoquinoline alkaloids (BIAs), have been shown inhibitory effects on pulmonary fibrosis (PF) through anti-inflammatory, anti-oxidative activities, inhibition of cytokines and NF-κB. Effects of other similar BIAs, Dauricine (Dau), Papaverine (Pap) and lotusine (Lot), on PF remain unclear. Here, we explored the effects of five bisbenzylisoquinoline (Lien, Nef, Iso, Tet and Dau) and two monobenzylisoquinoline (Pap, Lot) alkaloids on normal and PF fibroblasts.

METHODS

Primary normal and PF lung fibroblasts were cultured and treated with these alkaloids. Proliferation, activation, migration and apoptosis changes were detected by MTT, wound healing assay, flow cytometry. Protein level was analyzed by Western blot.

RESULTS

All BIAs inhibited proliferation of normal and PF lung fibroblasts induced by TGF-β. α-SMA protein level in normal and PF lung fibroblasts decreased after Lien, Nef, Iso, Tet and Dau treatment. Pap and Lot had no influence on α-SMA expression. Dau showed the strongest inhibitory effects on proliferation and activation among alkaloids. The migration rates of normal and PF lung fibroblasts were inhibited by Lien, Nef, Iso, and Dau. Lien, Nef, Iso and Dau significantly promoted apoptosis, while Tet had no effect on apoptosis. Pap and Lot had no influence on activation, migration and apoptosis. Dau significantly inhibited Smad3/4 and p-ERK1/2 protein overexpression induced by TGF-β1.

CONCLUSIONS

Bisbenzylisoquinoline alkaloids had stronger effects on inhibiting lung fibroblasts than monobenzylisoquinoline alkaloids. Dau expressed the strongest inhibitory effects, which may be related to its inhibition of TGF-β1/Smad3/4 and p-ERK1/2 pathway proteins.

背景

莲心碱（Lien）、甲基莲心碱（Nef）、异莲心碱（Iso）和粉防己碱（Tet），均为苄基异喹啉生物碱（BIAs），已证实可通过抗炎、抗氧化活性、抑制细胞因子和核因子κB对肺纤维化（PF）产生抑制作用。其他类似的BIAs，如蝙蝠葛碱（Dau）、罂粟碱（Pap）和荷叶碱（Lot）对PF的影响尚不清楚。在此，我们探究了五种双苄基异喹啉（莲心碱、甲基莲心碱、异莲心碱、粉防己碱和蝙蝠葛碱）和两种单苄基异喹啉（罂粟碱、荷叶碱）生物碱对正常和成纤维细胞性肺纤维化（PF）的影响。

方法

培养原代正常和成纤维细胞性肺纤维化肺成纤维细胞，并用这些生物碱进行处理。通过MTT法、伤口愈合试验、流式细胞术检测细胞增殖、活化、迁移和凋亡的变化。通过蛋白质印迹法分析蛋白质水平。

结果

所有BIAs均抑制转化生长因子-β（TGF-β）诱导的正常和成纤维细胞性肺纤维化肺成纤维细胞的增殖。莲心碱、甲基莲心碱、异莲心碱、粉防己碱和蝙蝠葛碱处理后，正常和成纤维细胞性肺纤维化肺成纤维细胞中的α-平滑肌肌动蛋白（α-SMA）蛋白水平降低。罂粟碱和荷叶碱对α-SMA表达无影响。在生物碱中，蝙蝠葛碱对增殖和活化的抑制作用最强。莲心碱、甲基莲心碱、异莲心碱和蝙蝠葛碱抑制正常和成纤维细胞性肺纤维化肺成纤维细胞的迁移率。莲心碱、甲基莲心碱、异莲心碱和蝙蝠葛碱显著促进细胞凋亡，而粉防己碱对细胞凋亡无影响。罂粟碱和荷叶碱对活化、迁移和凋亡无影响。蝙蝠葛碱显著抑制TGF-β1诱导的Smad3/4和磷酸化细胞外信号调节激酶1/2（p-ERK1/2）蛋白的过表达。