Hepatocellular Toxicity of Paris Saponins I, II, VI and VII on Two Kinds of Hepatocytes-HL-7702 and HepaRG Cells, and the Underlying Mechanisms

Abstract

Rhizoma paridis is a popularly-used Chinese medicine in clinics, based on the pharmacodynamic properties of its saponin components. The four main saponins in Rhizoma paridis are designated saponins I, II, VI, and VII. At present, much attention is focused on the anticancer effect of Rhizoma paridis which is manifested in its cytotoxicity to various cancer cells. The purpose of this study was to investigate the hepatocellular toxicities of the four saponins in Rhizoma paridis and the relative intensities of their cytotoxic effects. It was found that the four saponins were cytotoxic to two types of hepatocytes-HL-7702 and HepaRG cells. The cytotoxicities of the four saponins to the two cell models were compared. One of the most cytotoxic saponins was Rhizoma paridis saponin I (PSI). This was used to determine the mechanism of hepatocellular toxicity. Results from MTT assays demonstrated that the four saponins induced apoptosis of the two hepatocyte models in a dose-dependent and time-dependent manner. In addition, fluorescent 4′,6-diamidino-2-phenylindole (DAPI) staining was used to observe the morphological changes of HepaRG cells after saponin administration. Further, as the concentration increased, PSI-induced lactate dehydrogenase (LDH) release from HepaRG cells increased gradually. In addition, PSI enhanced the levels of reactive oxygen species (ROS) and blocked the S and G2 phases of the cell cycle in HepaRG cells. A western blot indicated that PSI upregulated the protein expression levels of p53, p21, and Fas. Furthermore, the PSI-induced changes in the p53 protein increased the Bax/bcl-2 ratio, resulting in enhancement of the release of mitochondrial cytochrome c, activation of caspases-3, -8, and -9, poly-ADP ribose polymerase (PARP), and ultimately apoptosis. Increased Fas protein activated caspase-8, which led to the activation of caspase-3 and its downstream PARP protein, resulting in cell apoptosis. These results indicate that PSI induced apoptosis in HepaRG cells through activation of ROS and death receptor pathways. The results obtained in this study suggest that the hepatocellular toxicity of saponins in Rhizoma paridis should be considered during the clinical application of this drug. In addition, they provide a reference for future anti-cancer studies on Rhizoma paridis.

Protective Effect of Flaxseed on the Health of Experimental Animals Exposed to Xylene

“…We determined the activity of LDH in the extracts of liver homogenates. The LDH is an intracellular enzyme that after release to the surrounding environment indicates damage to tissues [4,5,22,40]. The activity of the total lactate dehydrogenase in the liver sample extracts was higher in groups X and XF in comparison to the control group C, although the difference was insignificant (Fig.…”

Bioactive secondary metabolites in Paris polyphylla Sm. and their biological activities: A review

“…Similarly, paris saponin H inhibited the growth of liver cancer cells with an IC 50 of 1.25 μM (Chen et al, 2019), diosgenin lung cancer cells with IC 50 of 149.75 AE 10.43 μM (Yan et al, 2009), pennogenins liver cancer cells with IC 50 of 9.7-13.5 μM (Zhu et al, 2011). Paris saponin I inhibited the growth of ovarian cancer cells with IC 50 of <15 μM (Xiao et al, 2009), liver cancer cells with IC 50 of 0.84-4.66 μM (Wang et al, 2019), gastric cancer cells with IC 50 from 30.4 to 20.3 μM (Song et al, 2016) and lung cancer cells with IC 50 from 1.21 to 2.51 μg/mL (Jiang et al, 2014a(Jiang et al, , 2014bLiu et al, 2016;Zhao et al, 2015). Likewise, paris saponin II inhibited the growth of lung cancer cells (Zhang et al, 2016a,b), liver cancer cells (Wang et al, 2019), and ovary cancer cells (Xiao et al, 2012(Xiao et al, , 2014& Yang et al, 2015); polyphyllin I inhibited the growth of liver cancer cells (Xiao et al, 2018;Han et al, 2015), bone cancer cells (Chang et al, 2015) and lung cancer cells (Feng et al, 2019); polyphyllin VII inhibited the growth of lung cancer cells (Lin et al, 2015;He et al, 2020;Feng et al, 2019;Lin et al, 2015), liver cancer cells (Zhang et al, 2016a,b), nasopharyngeal cancer cells (Chen et al, 2016) and brain cancer cells (Pang et al, 2019;Liu et al, 2020).…”

“…Paris saponin VI showed anticancer activity toward the liver cancer line with IC 50 of 8.18 μM and 6.65 μM (Wang et al, 2019). Paris saponin VII inhibited the growth of human cervical cancer cells with an IC 50 of 2.62 AE 0.11 μM (Zhang et al, 2014), liver cancer cells with an IC 50 of 0.80-2.75 μM (Wang et al, 2019;Tang et al, 2019) and drug-resistant ovarian cancer cell lines (Yang et al, 2015a,b).…”

“…Paris saponin VI showed anticancer activity toward the liver cancer line with IC 50 of 8.18 μM and 6.65 μM (Wang et al, 2019). Paris saponin VII inhibited the growth of human cervical cancer cells with an IC 50 of 2.62 AE 0.11 μM (Zhang et al, 2014), liver cancer cells with an IC 50 of 0.80-2.75 μM (Wang et al, 2019;Tang et al, 2019) and drug-resistant ovarian cancer cell lines (Yang et al, 2015a,b). Similarly, paris saponin H inhibited the growth of liver cancer cells with an IC 50 of 1.25 μM (Chen et al, 2019), diosgenin lung cancer cells with IC 50 of 149.75 AE 10.43 μM (Yan et al, 2009), pennogenins liver cancer cells with IC 50 of 9.7-13.5 μM (Zhu et al, 2011).…”

Toxic Prediction of Pyrrolizidine Alkaloids and Structure‐Dependent Induction of Apoptosis in HepaRG Cells

“…HepaRG cells possess a hepatocyte-like morphology, maintaining hepatic functions and expression of liver-specific genes, including the expression of critical metabolism enzymes, drug transporters, and nuclear receptors, such as CYP enzymes, phase II enzymes, albumin, transferrin, aldolase B, and apical and canalicular ABC transporters and nuclear receptors (CAR, PXR) at levels comparable to human primary hepatocytes [ 19 , 20 ]. Due to these properties, the HepaRG cell line, with its metabolic competence and long-term cultivability, is a beneficial in vitro test system to assess the hepatocellular toxicity induced by plant extracts [ 21 , 22 ].…”