Effects of Geniposide and Geniposidic Acid on Fluoxetine-Induced Muscle Atrophy in C2C12 Cells

Fluoxetine, an antidepressant known as a selective 5-hydroxytryptamine reuptake inhibitor (SSRI), can cause side effects such as muscle atrophy with long-term use, but the mechanism is not fully understood. Geniposide (GPS) and geniposidic acid (GPSA), the main components of Gardenia jasminoides fruit, have been shown to have biological activity in disease prevention, but their role in preventing FXT-related side effects such as muscle atrophy remains unclear. The process of muscle atrophy is a complex physiological mechanism involving the balance of protein synthesis and catabolism. In this study, we hypothesized that FXT may suppress hypertrophy signaling and activate the atrophy mechanisms, resulting in proteolysis and reduced protein synthesis, while geniposide (GPS) and geniposide acid (GPSA) may be beneficial in improving muscle weakness caused by FXT. The C2C12 cell model was used to examine the expression of hypertrophy signaling (PI3K, Akt, and mTOR) and protein break signals (FOXO, MuRF-1, and MyHC). Our data indicated that FXT inhibited MyHC and promoted MuRF-1 protein expression by downregulating the signaling pathways of p-ERK1/2, p-Akt, p-mTOR, and p-FOXO, resulting in a decrease in differentiation and myotube formation in C2C12 muscle cells, which further resulted in muscle atrophy. However, GPS and GPSA can positively regulate the atrophy mechanism induced by FXT in muscle cells, thereby ameliorating the imbalance in muscle synthesis. In conclusion, GPS and GPSA have the potential to attenuate the muscle loss caused by long-term FXT administration, diseases, or the aging process.

A New Geniposidic Acid Derivative Exerts Antiaging Effects through Antioxidative Stress and Autophagy Induction

Two compounds that can prolong the replicative lifespan of yeast, geniposidic acid (Compound 1) and geniposide (Compound 2), were isolated from Gardenia jasminoides Ellis. Compared with Compound 1, Compound 2 was different at C11 and showed better bioactivity. On this basis, seven new geniposidic derivatives (3–9) were synthesized. Geniposidic 4-isoamyl ester (8, GENI), which remarkably prolonged the replicative and chronological lifespans of K6001 yeast at 1 µM, was used as the lead compound. Autophagy and antioxidative stress were examined to clarify the antiaging mechanism of GENI. GENI increased the enzymes activities and gene expression levels of superoxide dismutase (SOD) and reduced the contents of reactive oxygen species (ROS) and malondialdehyde (MDA) to improve the survival rate of yeast under oxidative stress. In addition, GENI did not extend the replicative lifespan of ∆sod1, ∆sod2, ∆uth1, ∆skn7, ∆cat, and ∆gpx mutants with K6001 background. The free green fluorescent protein (GFP) signal from the cleavage of GFP-Atg8 was increased by GENI. The protein level of free GFP showed a considerable increase and was time-dependent. Furthermore, GENI failed to extend the replicative lifespans of ∆atg32 and ∆atg2 yeast mutants. These results indicated that antioxidative stress and autophagy induction were involved in the antiaging effect of GENI.

Geniposidic Acid Confers Neuroprotective Effects in a Mouse Model of Alzheimer’s Disease through Activation of a PI3K/AKT/GAP43 Regulatory Axis

BACKGROUND: Alzheimer’s disease (AD) is a major cause of dementia, which is a growing global health problem and has a huge impact on individuals and society. As the modifying role of geniposidic acid (GPA) has been suggested in AD, this study sets out to determine if and how GPA treatment affects AD progression in mice. METHODS: Potential downstream target genes of GPA during AD were identified by bioinformatics analysis, revealing GAP43 as a primary candidate protein. Then, mPrP-APPswe/PS1De9 AD transgenic mice were treated with GPA via intragastric administration. This allowed for gain- and loss-of-function assays of candidate proteins being carried out with or without GPA treatment, after which behavioral tests could be conducted for mice. Cortical neuron apoptosis was measured by TUNEL staining, Amyloid β-protein (Aβ) expression in cerebral cortex by Thioflavin-s staining, and Aβ, IL-1β, IL-6, IL-4 and TNF-α levels in cerebral cortex by ELISA. GAP43 expression in cerebral cortex of mice was detected by immunohistochemistry. Primary cortical neurons of embryonic mice were isolated and induced by Aβ1-42 to construct AD cell model. Cell viability was assessed by CCK-8, and axon growth by immunofluorescence. RESULTS: GPA administration significantly improved the cognitive impairment, reducing Aβ accumulation and neuronal apoptosis in AD mice, and alleviated inflammation and axonal injury of Aβ1-42-induced neurons. GAP43 was shown experimentally to be the target of GPA in AD. Silencing of GAP43 repressed the neuroprotective effect of GPA treatment on AD mice. GPA elevated GAP43 expression via PI3K/AKT pathway activation and ultimately improved nerve injury in AD mice. CONCLUSION: GPA activates a PI3K/AKT/GAP43 regulatory axis to alleviate AD progression in mice.

Comparative Pharmacokinetics of Geniposidic Acid, Genipin-1-β-Gentiobioside, Geniposide, Genipin, and Crocetin in Rats after Oral Administration of Crude Gardeniae Fructus and Its Three Processed Products Using LC-MS/MS

The extract of Gardeniae Fructus (GF) with different processing methods processed the different medicinal properties and efficacy. Crude GF (CGF) could be processed into stir-frying GF (SGF), gancao mix-frying GF (GCGF), and ginger mix-frying GF (GIGF) in practice. An LC-MS/MS method was established for simultaneous quantification of geniposidic acid, geniposide, genipin-1-β-gentiobioside, genipin, and crocetin in the rat plasma. The LLOQs for determination of all five components were 10 ng/mL. The accuracies of intraday and interday were in the range of 91%–105%. The recoveries of 5 analytes ranged from 81.0% to 114% with RSD less than 14%. The results showed that the AUCs (area under the plasma concentration-time curve) and Cmax (maximum plasma concentration) of geniposidic acid, genipin-1-β-gentiobioside, and geniposide after oral administration of the CGF extract were apparently higher than those after oral administration of other processed extracts. Cmax of geniposide in plasma after administration of GIGF significantly decreased (p<0.01). Genipin was not detected in rat plasma after administration of the GIGF extract, but it can be detected in plasma after administration of CGF, SGF, and GCGF extract. Furthermore, crocin I and crocin II were not detected in plasma samples. Crocetin had higher concentration in rat plasma versus lower contents in extract. It was demonstrated that the different processing methods might influence the pharmacokinetics of geniposidic acid, genipin-1-β-gentiobioside, geniposide, genipin, and crocetin.