Naringenin inhibits APAP-induced acute liver injury through activating PPARA-dependent signaling pathway.

Acute liver injury (ALI) refers to the damage to the liver cells of patients due to drugs, food, and diseases. In this work, we used a network pharmacology approach to analyze the relevant targets and pathways of the active ingredients in Citri Reticulatae Pericarpium (CRP) for the treatment of ALI and conducted systematic validation through in vivo and in vitro experiments. The network pharmacologic results predicted that naringenin (NIN) was the main active component of CRP in the treatment of ALI. GO functional annotation and KEGG pathway enrichment showed that its mechanism may be related to the regulation of PPARA signaling pathway, PPARG signaling pathway, AKT1 signaling pathway, MAPK3 signaling pathway and other signaling pathways. The results of in vivo experiments showed that (NIN) could reduce the liver lesions, liver adipose lesions, hepatocyte injury and apoptosis in mice with APAP-induced ALI, and reduce the oxidative stress damage of mouse liver cells and the inflammation-related factors to regulate ALI. In vitro experiments showed that NIN could inhibit the proliferation, oxidative stress and inflammation of APAP-induced LO2 cells, promote APAP-induced apoptosis of LO2 cells, and regulate the expression of apoptotic genes in acute liver injury. Further studies showed that NIN inhibited APAP-induced ALI mainly by regulating the PPARA-dependent signaling pathway. In conclusion, this study provides a preliminary theoretical basis for the screening of active compounds in CRP for the prevention and treatment of ALI.

Identification of volatile and nonvolatile compounds in Citri Reticulatae Pericarpium Viride using GC-MS, UPLC-Q-Exactive Orbitrap-MS, and HPLC-PDA.

Gas chromatograph-mass spectrometer (GC-MS), ultra-high-performance liquid chromatograph-Q-Exactive Orbitrap tandem mass spectrometry (UHPLC-Q-Exactive Orbitrap-MS), and high-performance liquid chromatography-photodiode array detection (HPLC-PDA) were used to qualitatively and quantitatively analyze the chemical component of Citri Reticulatae Pericarpium Viride "Geqingpi" (GQP). First of all, the volatile components of GQP are identified by GC-MS. Totally 56 volatile components were determined, and γ-Terpinene (33.39%) and D-Limonene (22.95%) were the main terpenes. Secondly, UHPLC-Q-Exactive Orbitrap-MS was used for identifying nonvolatile compositions and 42 compositions were identified totally, including 23 flavonoids, nine organic acids, three coumarins, two alkaloids compounds, and five other compounds, among which nine of the determined constituents were detected for the first time in GQP. Thirdly, the content of seven main constituents in GQP was quantitatively analyzed via HPLC-PDA, which were synephrine, hesperidin, limonin, nobiletin, HMF, tangeretin, and 5-HPMF. Further investigation for quantitative analysis of seven bioactive compounds suggested that the concentration of hesperidin in GQP approximately was 16.0% (160.78 ± 0.95 mg·g-1), which was far higher than the standard for identification and quality control of CRPV in Chinese Pharmacopoeia (2020 edition) that "the content of hesperidin shall not be less than 5.0%." The phytochemicals of GQP were elucidated in this study, which might be supporting information for identification between GQP and Citri Reticulatae Pericarpium Viride "Sihuaqingpi" (SHQP) and provided a scientific basis for the further active ingredient for pharmacological research and development prospects of GQP.