Licochalcone A Induces Ferroptosis in Hepatocellular Carcinoma via Reactive Oxygen Species Activated by the SLC7A11/GPX4 Pathway.

Liver cancer is a common malignant tumor, and its incidence is increasing yearly. Millions of people suffer from liver cancer annually, which has a serious impact on global public health security. Licochalcone A (Lico A), an important component of the traditional Chinese herb licorice, is a natural small molecule drug with multiple pharmacological activities. In this study, we evaluated the inhibitory effects of Lico A on hepatocellular carcinoma cell lines (HepG2 and Huh-7), and explored the inhibitory mechanism of Lico A on hepatocellular carcinoma. First, we evaluated the inhibitory effects of Lico A on hepatocellular carcinoma, and showed that Lico A significantly inhibited and killed HepG2 and Huh-7 cells in vivo and in vitro. Transcriptomic analysis showed that Lico A inhibited the expression of solute carrier family 7 member 11 (SLC7A11), which induced ferroptosis. We confirmed through in vivo and in vitro experiments that Lico A promoted ferroptosis in hepatocellular carcinoma cells by downregulating SLC7A11 expression, thereby inhibiting the glutathione (GSH)-glutathione peroxidase 4 (GPX4) pathway and inducing activation of reactive oxygen species (ROS). In this study, we suggest that Lico A is a potential SLC7A11 inhibitor that induces ferroptotic death in hepatocellular carcinoma cells, thereby providing a theoretical basis for the development of natural small molecule drugs against hepatocellular carcinoma.

Potential mechanism of Ziyin Tongluo Formula in the treatment of postmenopausal osteoporosis: based on network pharmacology and ovariectomized rat model.

BACKGROUND: Amending from ancient classic, Ziyin Tongluo Formula (ZYTLF) has been prescribed to treat postmenopausal osteoporosis (PMOP) for decades with good curative effect. However, the possible mechanisms of it are still unknown. METHODS: Ovariectomized rat model was established to validate the therapeutic effect of ZYTLF on PMOP by Micro-CT bone analysis and pathological observation. Subsequently, active ingredients of ZYTLF and corresponding putative targets were identified by online databases. Overlapping genes were first obtained from mining genes associated with PMOP and then overlapped them with the putative targets. Key genes were selected from the multiple constructed and analyzed networks. GO and KEGG pathway enrichment analysis were performed by importing the key genes to the DAVID database. Moreover, validation of the binding association between key targets and their corresponding active compounds were accomplished by AutoDock Tools and other software. Lastly, Enzyme linked immunosorbent assay (Elisa) detection and Western blot analysis were utilized to further explore the possible mechanism of ZYTLF on PMOP. RESULTS: With 129 target genes interacting with PMOP, 92 active compounds of ZYTLF corresponded to 243 targets, and 50 key genes were chosen. Network analysis revealed the top 10 active ingredients, such as quercetin and kaempferol and the top 50 key genes, such as ERα, p38 MAPK, p-AKT and TGF-ß1. Enrichment analysis uncovered multiple signaling pathways, including estrogen signaling pathway, TNF signaling pathway, PI3K-Akt signaling pathway and MAPK signaling pathway. Furthermore, our finding of the foremost active compounds was tightly bound to the core proteins, which were verified by molecular docking analysis. Through experimental studies, we confirmed that the prescription of ZYTLF could ameliorate the OVX-induced bone loss, suppress the osteoclast activity and boost osteoblast ability through experimental studies. CONCLUSION: The potential mechanisms and therapeutic effects of ZYTLF against PMOP may be ascribed to inhibition of osteoclast activity, boost of osteoblast activity and enhancement of the expression of ERα.

Impact of the haplotypes of the human pregnane X receptor gene on the basal and St John's wort-induced activity of cytochrome P450 3A4 enzyme.

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT: Human pregnane X receptor (PXR/NR1I2) is a key regulator of cytochrome P450 3A4. To date, there are 198 reported SNPs for the human PXR/NR1I2 gene. Some of these SNPs are found to affect the inducing ability of PXR to CYP3A4. WHAT THIS STUDY ADDS: This study, for the first time, has investigated the effect of PXR haplotype on basal and St John's wort-induced CYP3A4 activity in humans. H1/H1 of the PXR gene had weaker basal transcriptional activity but greater inducible transcriptional activity to CYP3A4 than H1/H2 and H2/H2. AIMS: Human pregnane X receptor (PXR/NR1I2) is the master regulator of CYP3A4, which metabolizes >50% of drugs on the market. This study investigated the relationship between the two most frequent haplotypes [H1 (TCAGGGGCCACC) and H2 (CCGAAAACTAAT)] of PXR and basal and St John's wort (SJW)-induced CYP3A4 activity. METHODS: Ten healthy subjects carrying H1 and H2 haplotypes (three subjects with H1/H1, four with H1/H2 and three with H2/H2) entered this study. The 10 subjects did not carry CYP3A4*4, *5 and *6. All subjects were administrated a 300-mg SJW tablet three times daily for 14 days, and CYP3A4 activity was measured using nifedipine (NIF) as a probe. The plasma concentrations of NIF and dehydronifedipine (DNIF) were determined by a validated liquid chromatography/mass spectrometry/mass spectrometry method. RESULTS: After administration of SJW, the AUC(0-infinity) of NIF decreased significantly, and the AUC(0-infinity) of DNIF increased significantly (P < 0.05). For H1/H2, the AUC(0-infinity) of NIF decreased by 42.4%, and the AUC(0-infinity) of DNIF increased by 20.2%; for H2/H2, the AUC(0-infinity) of NIF decreased by 47.9%, and the AUC(0-infinity) of DNIF increased by 33.0%; for H1/H1, the AUC(0-infinity) of NIF decreased by 29.0%, yet the AUC(0-infinity) of DNIF increased by 106.7%. The increase of the AUC(0-infinity) of DNIF in H1/H1 was significantly different from the other two haplotype pairs (P < 0.05). Meanwhile, before administration of SJW, the ratio of AUC(0-infinity(DNIF))/AUC(0-infinity(NIF)) was the lowest for H1/H1 (22.1%), compared with H1/H2 (58.7%) and H2/H2 (30.0%). CONCLUSIONS: H1/H1 of the human PXR gene had weaker basal transcriptional activity but greater inducible transcriptional activity to CYP3A4 than H1/H2 and H2/H2.

Circadian variation of plasma cortisol and whole blood reduced glutathione levels in nasopharyngeal carcinoma patients.

BACKGROUND & OBJECTIVE: Glutathione is involved in cellular protection against radiation damage and drug detoxification. This study was to investigate the circadian variation of plasma cortisol and whole blood reduced glutathione (GSH) levels in nasopharyngeal carcinoma (NPC) patients to provide references for chronotherapy for NPC. METHODS: A total of 13 NPC patients and 14 healthy volunteers were involved. Peripheral venous blood was sampled every 4 h during one 24-hour period starting at 12:00 am. The plasma cortisol concentration was determined by radioimmunoassay; the GSH concentration was determined by high performance liquid chromatography. RESULTS: Plasma cortisol levels of both groups displayed clear and similar circadian rhythms. The cortisol level peaked in both groups in the morning and was the lowest at mid-night. In both groups, GSH concentrations showed significant differences according to sampling time (ANOVA for Repeated Measures, F=5.18, P=0.02). By cosinor analysis, the circadian variation of the GSH level in NPC group was marginally statistically significant (Cosinor analysis, P=0.06) with the acrophase appeared at 05:02; the GSH level in control group displayed an obvious circadian rhythm and the acrophase appeared at 07:44+/-01:56 (P<0.01). The rhythm-adjust mean value of glutathione was (19.60+/-1.11) nmol/mg protein in NPC group and (8.95+/-0.46) nmol/mg protein in control group. CONCLUSIONS: The circadian rhythm of the plasma cortisol level is maintained in NPC patients, even in some patients at advanced stages. In NPC patients, GSH shows a trend of circadian variation which is similar to that in healthy controls. These results could be used to select special schedules for chrono-radiotherapy and chrono-chemotherapy for NPC patients.