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Effect of the PGD2-DP signaling pathway on primary cultured rat hippocampal neuron injury caused by aluminum overload.

Ma J, Yang Q, Wei Y, Yang Y, Ji C, Hu X, Mai S, Kuang S, Tian X, Luo Y, Liang G, Yang J - Sci Rep (2016)

Bottom Line: BW245C reduced the Ca(2+) fluorescence intensity and protected the neurons.DK-PGD2 increased the intensity of Ca(2+) fluorescence, while CAY10471 had the opposite effect.In conclusion, contrary to the effect of DP2, the PGD2-DP1 signaling pathway protects against the primary cultured rat hippocampal neuronal injury caused by aluminum overload.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Chongqing Medical University, the Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing 400016, China.

ABSTRACT
In the present study, the agonists and antagonists of DP receptor were used to examine whether the PGD2-DP signaling pathway affects neuronal function. Primary cultured hippocampal neuron was prepared and treated with aluminum maltolate (100 μM) to establish the neuronal damage model. PGD2 and cAMP content was detected by ELISA. L-PGDS and DPs mRNA and protein expression were measured by RT-PCR and Western blotting, respectively. The aluminium-load neuron was treated with the DP1 agonist BW245C, the DP1 antagonist BWA868C, the DP2 agonist DK-PGD2, and the DP2 antagonist CAY10471, respectively. Neuronal pathomorphology was observed using H-E staining. The cell viability and the lactate dehydrogenase leakage rates of neurons were measured with MTT and LDH kit, respectively. Ca(2+) level was detected by Fluo-3/AM. In the model group, the MTT values obviously decreased; LDH leakage rates and PGD2 content increased significantly; L-PGDS, DP1 mRNA and protein expressions increased, and DP2 level decreased. BW245C reduced the Ca(2+) fluorescence intensity and protected the neurons. DK-PGD2 increased the intensity of Ca(2+) fluorescence, while CAY10471 had the opposite effect. In conclusion, contrary to the effect of DP2, the PGD2-DP1 signaling pathway protects against the primary cultured rat hippocampal neuronal injury caused by aluminum overload.

No MeSH data available.


Related in: MedlinePlus

Effects of DP agonists and antagonists on the changes of neuronal viability caused by aluminium detected by the method of MTT.(a) BW245C and (d) CAY10471 increased neuron viability in a concentration dependent manner in Al3+-treated group, whereas (b) BWA868C and (c) DK-PGD2 decreased neuron viability in Al3+-treated group. Values were mean ± SD of eight individual experiments (n = 8, ##P < 0.01 compared with control group, *P < 0.05 and **P < 0.01 compared with Al3+-treated group, respectively, one-way ANOVA with Dunnett’s multiple comparisons).
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f5: Effects of DP agonists and antagonists on the changes of neuronal viability caused by aluminium detected by the method of MTT.(a) BW245C and (d) CAY10471 increased neuron viability in a concentration dependent manner in Al3+-treated group, whereas (b) BWA868C and (c) DK-PGD2 decreased neuron viability in Al3+-treated group. Values were mean ± SD of eight individual experiments (n = 8, ##P < 0.01 compared with control group, *P < 0.05 and **P < 0.01 compared with Al3+-treated group, respectively, one-way ANOVA with Dunnett’s multiple comparisons).

Mentions: About the concentrations of the agonists and antagonists of DP, for the observation of neuronal viability, seven concentrations (10−5, 3 × 10−5, 10−6, 3 × 10−6, 10−7, 3 × 10−7, 10−8, 3 × 10−8 M) of each agonists and antagonists of DP were used. The results showed that compared with the control group, the neuron viability decreased significantly in model group (P < 0.01) and that treatment of DP1 agonists (BW245C) at a concentration of 10−6, 3 × 10−5 and 10−5 M increased significantly the neuron viability compared with model group, whereas the viability of neuron treated with DP1 antagonist (BWA868C) of 10−5 M was decreased significantly (P < 0.01). Compared with model group, the neuron viability in the DP2 antagonist (CAY10471) (10−6, 3 × 10−5 and 10−5 M) -treated group was increased substantial. The administration of DP2 agonists (DK-PGD2) at a concentration of 10−5 and 3 × 10−5 M decreased neuron viability significantly (P < 0.01). There was no considerable difference between the control group and the solvent control group (Fig. 5).


Effect of the PGD2-DP signaling pathway on primary cultured rat hippocampal neuron injury caused by aluminum overload.

Ma J, Yang Q, Wei Y, Yang Y, Ji C, Hu X, Mai S, Kuang S, Tian X, Luo Y, Liang G, Yang J - Sci Rep (2016)

Effects of DP agonists and antagonists on the changes of neuronal viability caused by aluminium detected by the method of MTT.(a) BW245C and (d) CAY10471 increased neuron viability in a concentration dependent manner in Al3+-treated group, whereas (b) BWA868C and (c) DK-PGD2 decreased neuron viability in Al3+-treated group. Values were mean ± SD of eight individual experiments (n = 8, ##P < 0.01 compared with control group, *P < 0.05 and **P < 0.01 compared with Al3+-treated group, respectively, one-way ANOVA with Dunnett’s multiple comparisons).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835855&req=5

f5: Effects of DP agonists and antagonists on the changes of neuronal viability caused by aluminium detected by the method of MTT.(a) BW245C and (d) CAY10471 increased neuron viability in a concentration dependent manner in Al3+-treated group, whereas (b) BWA868C and (c) DK-PGD2 decreased neuron viability in Al3+-treated group. Values were mean ± SD of eight individual experiments (n = 8, ##P < 0.01 compared with control group, *P < 0.05 and **P < 0.01 compared with Al3+-treated group, respectively, one-way ANOVA with Dunnett’s multiple comparisons).
Mentions: About the concentrations of the agonists and antagonists of DP, for the observation of neuronal viability, seven concentrations (10−5, 3 × 10−5, 10−6, 3 × 10−6, 10−7, 3 × 10−7, 10−8, 3 × 10−8 M) of each agonists and antagonists of DP were used. The results showed that compared with the control group, the neuron viability decreased significantly in model group (P < 0.01) and that treatment of DP1 agonists (BW245C) at a concentration of 10−6, 3 × 10−5 and 10−5 M increased significantly the neuron viability compared with model group, whereas the viability of neuron treated with DP1 antagonist (BWA868C) of 10−5 M was decreased significantly (P < 0.01). Compared with model group, the neuron viability in the DP2 antagonist (CAY10471) (10−6, 3 × 10−5 and 10−5 M) -treated group was increased substantial. The administration of DP2 agonists (DK-PGD2) at a concentration of 10−5 and 3 × 10−5 M decreased neuron viability significantly (P < 0.01). There was no considerable difference between the control group and the solvent control group (Fig. 5).

Bottom Line: BW245C reduced the Ca(2+) fluorescence intensity and protected the neurons.DK-PGD2 increased the intensity of Ca(2+) fluorescence, while CAY10471 had the opposite effect.In conclusion, contrary to the effect of DP2, the PGD2-DP1 signaling pathway protects against the primary cultured rat hippocampal neuronal injury caused by aluminum overload.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Chongqing Medical University, the Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing 400016, China.

ABSTRACT
In the present study, the agonists and antagonists of DP receptor were used to examine whether the PGD2-DP signaling pathway affects neuronal function. Primary cultured hippocampal neuron was prepared and treated with aluminum maltolate (100 μM) to establish the neuronal damage model. PGD2 and cAMP content was detected by ELISA. L-PGDS and DPs mRNA and protein expression were measured by RT-PCR and Western blotting, respectively. The aluminium-load neuron was treated with the DP1 agonist BW245C, the DP1 antagonist BWA868C, the DP2 agonist DK-PGD2, and the DP2 antagonist CAY10471, respectively. Neuronal pathomorphology was observed using H-E staining. The cell viability and the lactate dehydrogenase leakage rates of neurons were measured with MTT and LDH kit, respectively. Ca(2+) level was detected by Fluo-3/AM. In the model group, the MTT values obviously decreased; LDH leakage rates and PGD2 content increased significantly; L-PGDS, DP1 mRNA and protein expressions increased, and DP2 level decreased. BW245C reduced the Ca(2+) fluorescence intensity and protected the neurons. DK-PGD2 increased the intensity of Ca(2+) fluorescence, while CAY10471 had the opposite effect. In conclusion, contrary to the effect of DP2, the PGD2-DP1 signaling pathway protects against the primary cultured rat hippocampal neuronal injury caused by aluminum overload.

No MeSH data available.


Related in: MedlinePlus