Limonin ameliorates cisplatin-induced acute liver injury by inhibiting 11β-hydroxysteroid dehydrogenase type 1.
作者: Yadie Xiang ; Xianke Zhou ; Hong Zhou ; Dier Li ; Menghua Zhong ; Xue Hong ; Dongyan Song ; Yinyi Long ; Xi Zeng ; Yudan Chen ; Jiayi Zhou ; Dongning Liang ; Haiyan Fu
Acute liver injury (ALI) is a common side effect of cisplatin treatment in the clinic and can lead to liver failure if not treated promptly. Previous studies have revealed that Limonin, a critical bioactive substance in citrus fruits, can protect multiple organs from various medical conditions. However, whether Limonin could ameliorate cisplatin-induced ALI remains unclear.
In vivo and in vitro models were induced by cisplatin in the present study. Non-targeted metabolomics was employed to analyze the metabolic changes in the liver after ALI. In addition, molecular docking was utilized to predict the potential targets of Limonin.
Limonin attenuated hepatic histopathological injury by reducing hepatocyte apoptosis, lipid peroxidation, and inflammation in cisplatin-challenged mice. Employing metabolomics, we revealed that Limonin mediated the balance of various disturbed metabolic pathways in the liver after cisplatin-induced ALI. Integrating public data mining, molecular docking studies, and in vitro experiments demonstrated that Limonin suppressed the expression and activity of its direct target, 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), in the liver, thus reducing the production of corticosterone (CORT), a key metabolite promoted hepatocyte apoptosis.
Limonin improves the liver metabolic microenvironment by inhibiting 11β-HSD1 to protect against cisplatin-induced ALI.
2023-09-07·Plants (Basel, Switzerland)
Selection of Reference Genes in Evodia rutaecarpa var. officinalis and Expression Patterns of Genes Involved in Its Limonin Biosynthesis.
作者: Yu Zhou ; Yuxiang Zhang ; Detian Mu ; Ying Lu ; Wenqiang Chen ; Yao Zhang ; Ruiying Zhang ; Ya Qin ; Jianhua Yuan ; Limei Pan ; Qi Tang
E. rutaecarpa var. officinalis is a traditional Chinese medicinal plant known for its therapeutic effects, which encompass the promotion of digestion, the dispelling of cold, the alleviation of pain, and the exhibition of anti-inflammatory and antibacterial properties. The principal active component of this plant, limonin, is a potent triterpene compound with notable pharmacological activities. Despite its significance, the complete biosynthesis pathway of limonin in E. rutaecarpa var. officinalis remains incompletely understood, and the underlying molecular mechanisms remain unexplored. The main purpose of this study was to screen the reference genes suitable for expression analysis in E. rutaecarpa var. officinalis, calculate the expression patterns of the genes in the limonin biosynthesis pathway, and identify the relevant enzyme genes related to limonin biosynthesis. The reference genes play a pivotal role in establishing reliable reference standards for normalizing the gene expression data, thereby ensuring precision and credibility in the biological research outcomes. In order to identify the optimal reference genes and gene expression patterns across the diverse tissues (e.g., roots, stems, leaves, and flower buds) and developmental stages (i.e., 17 July, 24 August, 1 September, and 24 October) of E. rutaecarpa var. officinalis, LC-MS was used to analyze the limonin contents in distinct tissue samples and developmental stages, and qRT-PCR technology was employed to investigate the expression patterns of the ten reference genes and eighteen genes involved in limonin biosynthesis. Utilizing a comprehensive analysis that integrated three software tools (GeNorm ver. 3.5, NormFinder ver. 0.953 and BestKeeper ver. 1.0) and Delta Ct method alongside the RefFinder website, the best reference genes were selected. Through the research, we determined that Act1 and UBQ served as the preferred reference genes for normalizing gene expression during various fruit developmental stages, while Act1 and His3 were optimal for different tissues. Using Act1 and UBQ as the reference genes, and based on the different fruit developmental stages, qRT-PCR analysis was performed on the pathway genes selected from the "full-length transcriptome + expression profile + metabolome" data in the limonin biosynthesis pathway of E. rutaecarpa var. officinalis. The findings indicated that there were consistent expression patterns of HMGCR, SQE, and CYP450 with fluctuations in the limonin contents, suggesting their potential involvement in the limonin biosynthesis of E. rutaecarpa var. officinalis. This study lays the foundation for further research on the metabolic pathway of limonin in E. rutaecarpa var. officinalis and provides reliable reference genes for other researchers to use for conducting expression analyses.
2023-08-24·Applied microbiology and biotechnology
Immobilization of Rhodococcus by encapsulation and entrapment: a green solution to bitter citrus by-products.
作者: María C Pilar-Izquierdo ; María López-Fouz ; Natividad Ortega ; María D Busto
Debittering of citrus by-products is required to obtain value-added compounds for application in the food industry (e.g., dietary fiber, bioactive compounds). In this work, the immobilization of Rhodococcus fascians cells by encapsulation in Ca-alginate hollow beads and entrapment in poly(vinyl alcohol)/polyethylene glycol (PVA/PEG) cryogels was studied as an alternative to chemical treatments for degrading the bitter compound limonin. Previously, the Rhodococcus strain was adapted using orange peel extract to increase its tolerance to limonoids. The optimal conditions for the encapsulation of microbial cells were 2% Na-alginate, 4% CaCl2, 4% carboxymethylcellulose (CMC), and a microbial load of 0.6 OD600 (optical density at 600 nm). For immobilization by entrapment, the optimal conditions were 8% PVA, 8% PEG, and 0.6 OD600 microbial load. Immobilization by entrapment protected microbial cells better than encapsulation against the citrus medium stress conditions (acid pH and composition). Thus, under optimal immobilization conditions, limonin degradation was 32 and 28% for immobilization in PVA/PEG gels and in hollow beads, respectively, in synthetic juice (pH 3) after 72 h at 25 °C. Finally, the microbial cells entrapped in the cryogels showed a higher operational stability in orange juice than the encapsulated cells, with four consecutive cycles of reuse (runs of 24 h at 25 °C). KEY POINTS: • Increased tolerance to limonoids by adapting R. fascians with citrus by-products. • Entrapment provided cells with favorable microenvironment for debittering at acid pH. • Cryogel-immobilized cells showed the highest limonin degradation in citrus products.