Cornus officinalis with high pressure wine steaming enhanced anti-hepatic fibrosis: Possible through SIRT3-AMPK axis

Cornus officinalis,a medicinal and edible plant known for its liver-nourishing properties,has shown promise in inhibiting the activation of hepatic stellate cells(HSCs),crucial indicators of hepatic fibrosis,especially when processed by high pressure wine steaming(HPWS).Herein,this study aims to investigate the regulatory effects of cornus officinalis,both in its raw and HPWS forms,on inflammation and apoptosis in liver fibrosis and their underlying mechanisms.In vivo liver fibrosis models were established by subcutaneous injection of CCl4,while in vitro HSCs were exposed to transforming growth factor-b(TGF-b).These findings demonstrated that cornus officinalis with HPWS conspicuously ameliorated histopathological injury,reduced the release of proinflammatory factors,and decreased collagen deposition in CCl4-induced rats compared to its raw form.Utilizing ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometer(UHPLC-QTOF-MS)combined with network analysis,we identified that the pharmacological effects of the changed components of cornus officinalis before and after HPWS,primarily centered on the adenosine phosphate(AMP)-activated protein kinase(AMPK)pathway.Of note,cornus officinalis activated AMPK and sirtuin 3(SIRT3),promoting the apoptosis of activated HSCs through the caspase cascade by regulating caspase3,caspase6 and caspase9.small interfering RNA(siRNA)experiments showed that cornus officinalis could regulate AMPK activity and its mediated-apoptosis through SIRT3.In conclusion,cornus officinalis exhibited the ability to reduce inflammation and apoptosis,with the SIRT3-AMPK signaling pathway identified as a potential mechanism underlying the synergistic effect of cornus officinalis with HPWS on anti-liver fibrosis.

桂七、台农芒果皮水提液对人肝癌细胞株HepG2的凋亡作用及其机制研究

目的研究桂七、台农芒果皮水提液对人肝癌细胞株HepG2的促凋亡作用及机制。方法将未加芒果皮水提液的人肝癌细胞株HepG2作为对照组,加4、8、12、16、20μg/μl芒果皮水提液的人肝癌细胞株HepG2为实验组,采用CCK-8法检测桂七、台农芒果皮水提液对人肝癌细胞株HepG2增殖的抑制作用,确定药物作用的最佳浓度;流式细胞仪测定桂七、台农芒果皮水提液作用前后的细胞凋亡率;酶联免疫吸附试验法检测细胞凋亡相关因子Caspase 6、Caspase 9含量。结果桂七芒果皮水提液作用于人肝癌细胞株HepG224 h后的最佳浓度为16μg/μl;台农芒果皮水提液作用于人肝癌细胞株HepG224 h后的最佳浓度为8μg/μl。桂七芒果皮水提液组、台农芒果皮水提液组细胞凋亡率高于对照组(P<0.05)。桂七芒果皮水提液组Caspase 6含量低于对照组;桂七芒果皮水提液组、台农芒果皮水提液组Caspase 9含量高于对照组(P<0.05)。结论桂七、台农芒果皮水提液对人肝癌细胞株HepG2有促凋亡作用,其机制可能与Caspase 6含量下降、Caspase 9含量上升有关。

Inhibitory Effects of INF-α and Curcumin on the Proliferation of Raji Cells

Objective： To investigate the inhibitory effect of interferon-α （IFN-α） and curcumin on proliferation of Raji cells （B-NHL） and its mechanism. Methods： The morphological, changes of Raji cells were observed in culture medium with IFN-α （500, 1000, 2000, 3000 U/L） and various concentrations of curcumin （6.25, 12.5, 25 μmol/L） for different time in vitro. The inhibitory ratio was measured by MTT assay. Apoptosis was detected by flow cytometry （FCM）. The expression of caspase 6, caspase 8 and caspase 9 in Raji cells treated with IC5025 μmol/L curcumin with IFN-α was examined using Western blot. Results： IFN-α and curcumin could significantly inhibit the growth and induce apoptosis of RAji cells with synergistic effects. They could increase the expression of caspase 6, caspase 8 and caspase 9 in Raji cells in a dose- and time-dependent manner. Conclusion： The combined use of IFN-α and curcumin can inhibit the proliferation of B-NHL Raji cells apparently in vitro. Promotion of the expression of caspase 6, caspase 8, caspase 9 and induction of apoptosis might be one of the important mechanisms.

干扰素-α联合姜黄素对B-NHL淋巴瘤Raji细胞的增生抑制作用

背景与目的:姜黄素(curcumin)是中药姜黄的主要成分,其选择性的抑制肿瘤细胞增殖并诱导其凋亡。干扰素-α是目前用于临床治疗最重要的细胞因子之一。本研究旨在探讨干扰素-α联合姜黄素对B-NHL淋巴瘤Raji细胞的诱导凋亡作用及其作用机制。方法:以浓度2000u/L干扰素-α分别与6.25、12.5、25μmol/L浓度的姜黄素作用于体外培养的Raji细胞,观察细胞生长状态变化。MTT法检测细胞生长的抑制率,应用流式细胞仪法观察细胞凋亡,Western blot法检测24h,姜黄素(IC5025μmol/L)以及联合干扰素-α(2000u/L)对caspase6、caspase8、caspase9蛋白的影响。结果:干扰素-α与姜黄素联合应用可显著抑制Raji细胞的生长,诱导细胞发生凋亡,两者具有协同效应。凋亡蛋白caspase6、caspase8、caspase9表达显著增高,其作用呈现出明显的量效与时效关系。结论:干扰素-α与姜黄素联合应用对B-NHL淋巴瘤Raji细胞具有明显的增生抑制作用;促进凋亡蛋白的表达及诱导细胞凋亡,可能是其重要作用机制之一。