Xiao-Feng Yan, Pei Zhao, Xiao-Ling Wang, Department of Biology, School of Basic Medical Science, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
World J Gastroenterol. 2017 Aug 7;23(29):5333-5344. doi: 10.3748/wjg.v23.i29.5333.
To investigate the capability of salvianolic acid B (Sal B) to protect hepatocytes from hydrogen peroxide (HO)/carbon tetrachloride (CCl)-induced lysosomal membrane permeabilization.
Cell Counting Kit-8 assay was used to measure cell viability. Apoptosis and death were assayed through flow cytometry. BrdU incorporation was used to detect cell proliferation. Serum alanine aminotransferase activity and liver malondialdehyde (MDA) content were measured. Liver histopathological changes were evaluated using hematoxylin-eosin staining. Lysosomal membrane permeability was detected with LysoTracker Green-labeled probes and acridine orange staining. The levels of protein carbonyl content (PCC), cathepsins (Cat)B/D, and lysosome-associated membrane protein 1 (LAMP1) were evaluated through western blotting. Cytosol CatB activity analysis was performed with chemiluminescence detection. The mRNA level of LAMP1 was evaluated through quantitative real-time polymerase chain reaction.
Results indicated that HO induced cell injury/death. Sal B attenuated HO-induced cell apoptosis and death, restored the inhibition of proliferation, decreased the amount of PCC, and stabilized the lysosome membrane by increasing the LAMP1 protein level and antagonizing CatB/D leakage into the cytosol. CCl also triggered hepatocyte death. Furthermore, Sal B effectively rescued hepatocytes by increasing LAMP1 expression and by reducing lysosomal enzyme translocation to the cytosol.
Sal B protected mouse embryonic hepatocytes from HO/CCl-induced injury/death by stabilizing the lysosomal membrane.
研究丹酚酸 B（Sal B）保护肝细胞免受过氧化氢（HO）/四氯化碳（CCl）诱导的溶酶体膜通透性的能力。
使用细胞计数试剂盒-8 法测定细胞活力。通过流式细胞术检测细胞凋亡和死亡。BrdU 掺入法检测细胞增殖。测定血清丙氨酸氨基转移酶活性和肝丙二醛（MDA）含量。采用苏木精-伊红染色评估肝组织病理学变化。用 LysoTracker Green 标记探针和吖啶橙染色检测溶酶体膜通透性。通过 Western 印迹评估蛋白羰基含量（PCC）、组织蛋白酶（Cat）B/D 和溶酶体相关膜蛋白 1（LAMP1）的水平。通过化学发光检测分析胞质 CatB 活性。通过实时定量聚合酶链反应评估 LAMP1 的 mRNA 水平。
结果表明，HO 诱导细胞损伤/死亡。Sal B 减轻了 HO 诱导的细胞凋亡和死亡，恢复了增殖抑制，通过增加 LAMP1 蛋白水平和拮抗 CatB/D 漏入胞质来稳定溶酶体膜，降低 PCC 含量。CCl 也引发了肝细胞死亡。此外，Sal B 通过增加 LAMP1 表达和减少溶酶体酶向胞质转移，有效地挽救了肝细胞。
Sal B 通过稳定溶酶体膜来保护小鼠胚胎肝细胞免受 HO/CCl 诱导的损伤/死亡。
