Synergistic neuroprotective effect of microglial-conditioned media treated with geniposide and ginsenoside Rg1 on hypoxia injured neurons

Abstract

The synergistic mechanism underlying the effects of multi‑component combined drug use for complex diseases remains to be fully elucidated. Microglial activation following ischemia can either affect neural survival or cause neuronal injury. The aim of the present study was to determine the synergistic effect of geniposide and ginsenoside Rg1, based on microglial‑neuronal communication. N2a neuronal cells were divided into the following seven groups: Control group; normal cultured microglial cells in conditioned medium (N‑MG‑CM) group; oxygen‑glucose deprivation (OGD) model group; OGD‑injured MG‑CM (I‑MG‑CM) group; geniposide‑treated MG‑CM (G‑MG‑CM) group; ginsenoside Rg1‑treated MG‑CM (R‑MG‑CM) group; and combination‑treated MG‑CM (C‑MG‑CM) group. A series of assays were used to detect the effects of the different MG‑CM on neurons in terms of: (i) cell viability, determined using a Cell Counting Kit‑8; (ii) lactate dehydrogenase (LDH) leakage rate; (iii) expression of NMDAR1 and activated caspase‑3, detected using western blotting; (iv) mitochondrial transmembrane potential, determined by JC‑1; and (v) mitochondrial ultrastructural features, determined using electron microscopy. The experimental results demonstrated that MG‑CM including the integrated use of geniposide and ginsenoside Rg1 significantly protected neuronal cell viability and inhibited LDH leakage, suppressed the expression of N‑methyl‑D‑aspartate receptor subunit 1 and activated caspase‑3, increased the mitochondrial transmembrane potential and improved the mitochondrial ultrastructure. MG‑CM from separately used geniposide or ginsenoside Rg1 demonstrated differential neuroprotection at different levels. These findings revealed that the synergistic drug combination of geniposide and ginsenoside Rg1 in the treatment of stroke is a feasible approach for use.

Protective Effects and Target Network Analysis of Ginsenoside Rg1 in Cerebral Ischemia and Reperfusion Injury: A Comprehensive Overview of Experimental Studies

“…Furthermore, after CI/RI, ATP production is insufficient and Na-K-ATPase activity is decreased, resulting in increased intracellular Na + concentrations and consequential cytotoxic cerebral edema and nerve damage [5]. After G-Rg1 treatment, the levels of ATP and adenosine monophosphate (AMP) markedly increased, the levels of total adenine nucleotides (TANs) and energy charge (EC) improved [96], and mitochondrial transmembrane potential (MMP) increased [97]. The mechanisms underlying these effects might be associated with improving mitochondrial ultrastructure and oxidative respiratory function, which lead to the inhibition of mitochondrial apoptosis, enhance the expression of glucose transporter 3 (GLUT3), promote the activation of AMPKα1/2, increase the uptake of glucose into nerve cells, and increase the supply and intake of glucose, as shown in Table 4.…”

Synergistic neuroprotective effects of Geniposide and ursodeoxycholic acid in hypoxia‑reoxygenation injury in SH‑SY5Y cells

“…Pharmacodynamic constituents from natural medicines have been investigated for the treatment of ischemic stroke. Multi-component treatments, characterized by the combination of two or more agents that interact with multiple targets simultaneously, are considered to be rational and efficient forms of therapy for the treatment of complex diseases ( 19 ). According to the results obtained from the CCK-8 and LDH assays, proteins secreted from OGD/R-activated SH-SY5Y cells are involved in the neuronal cell damage observed following treatment, indicating that OGD/R induces neurotoxicity in SH-SY5Y cells.…”

“…Total RNA was extracted from cells lysed using TRizol reagent (Takara Bio, Dalian, China) per the manufacturer's instructions ( 19 ). The quality and quantity of the RNA purity were assessed using a spectrophotometer and standard electrophoresis.…”

“…The neuroprotective effects of Geniposide have been well established, and our previous studies have shown that the effective components of Bezoar alleviate cerebral ischemic injury by inhibiting ERS ( 5 ). Moreover, TUDCA has been shown to be more effective than taurine and UDCA both in vivo and in vitro ( 19 ). However, the synergistic effects of the two components combined remains to be fully elucidated.…”

Notoginseng Leaf Triterpenes Ameliorates OGD/R-Induced Neuronal Injury via SIRT1/2/3-Foxo3a-MnSOD/PGC-1α Signaling Pathways Mediated by the NAMPT-NAD Pathway

“…Depletion of ATP often triggers the ischemic cascades such as membrane ion pump failure, efflux of cellular potassium, an influx of sodium, chloride, and membrane depolarization [ 11 – 13 ]. These mitochondrial disorders may trigger mitochondrial quality control, recover mitochondrial morphology [ 14 , 15 ], and further aggravate the multiple pathological progresses of cerebral I/R injury, including excitotoxicity, mitochondrial response, free radical release, acidotoxity, protein misfolding, and inflammation [ 3 – 5 , 16 ]. Thus, mitochondrial metabolic disorder of energy is seen as one of the hallmarks of I/R-induced neuronal death.…”