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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

The effects of the DP agonists and antagonists on primary cultured rat hippocampal neurons.(A)The picture showed the number and structure of neurons in (a) control group and (b) Al3+-treated group. (c) BW245C- treated group and (f) CAY10471-treated group raised the number and structure of neurons in model group, whereas (d) BWA868C-treated group and (e) DK-PGD2- treated group decreased the number and structure of neurons in the Al3+-treated group. Sections were pictured at ×400. (B) The intervention effect of DP on the cAMP level in the Al3+-treated group. Values were mean ± SD of six individual experiments. (n = 6, ##P < 0.01 vs. control group. *P < 0.05 vs. Al-treated group, one-way ANOVA with Dunnett’s multiple comparisons) (C) The picture showed the Ca2+ fluorescence of neurons in (a) control group and (b) model group, (c) BW245C and (f) CAY10471 decreased the Ca2+ fluorescence of neurons in model group. (d) BWA868C and (e) DK-PGD2 increased the Ca2+ fluorescence compared with model group (×400). (D) Summary graph showed the intensity of Ca2+ fluorescence relative levels in DP agonists and antagonists groups against model group. Values were mean ± SD of ten individual experiments (n = 10. ##P < 0.01 compared with control group. **P < 0.01 compared with Al3+-treated group, one-way ANOVA with Dunnett’s multiple comparisons).
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f7: The effects of the DP agonists and antagonists on primary cultured rat hippocampal neurons.(A)The picture showed the number and structure of neurons in (a) control group and (b) Al3+-treated group. (c) BW245C- treated group and (f) CAY10471-treated group raised the number and structure of neurons in model group, whereas (d) BWA868C-treated group and (e) DK-PGD2- treated group decreased the number and structure of neurons in the Al3+-treated group. Sections were pictured at ×400. (B) The intervention effect of DP on the cAMP level in the Al3+-treated group. Values were mean ± SD of six individual experiments. (n = 6, ##P < 0.01 vs. control group. *P < 0.05 vs. Al-treated group, one-way ANOVA with Dunnett’s multiple comparisons) (C) The picture showed the Ca2+ fluorescence of neurons in (a) control group and (b) model group, (c) BW245C and (f) CAY10471 decreased the Ca2+ fluorescence of neurons in model group. (d) BWA868C and (e) DK-PGD2 increased the Ca2+ fluorescence compared with model group (×400). (D) Summary graph showed the intensity of Ca2+ fluorescence relative levels in DP agonists and antagonists groups against model group. Values were mean ± SD of ten individual experiments (n = 10. ##P < 0.01 compared with control group. **P < 0.01 compared with Al3+-treated group, one-way ANOVA with Dunnett’s multiple comparisons).

Mentions: The structures of primary cultured hippocampal neurons were clear, complete and the protrusions of neurons were interwoven into the meshes in the control group. Compared with the control group, the number of neurons was substantial decreased and the protrusions were degenerated in the Al3+-treated group. Compared with the Al3+-treated group, the number of neurons increased and the structures recovered in BW245C-treated group. Treated with BWA868C, the neuronal injury was further aggravated compared with the Al3+-treated group. Treated with DK-PGD2, the hippocampal neurons almost exhibited karyopyknosis and disruption, whereas the bodies and nucleus of neurons were still clear and karyopyknosis compared with the Al3+-treated group. The neuronal injury in the Al3+-treated group was considerable reduced when treated with CAY10471 (Fig. 7A).


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)

The effects of the DP agonists and antagonists on primary cultured rat hippocampal neurons.(A)The picture showed the number and structure of neurons in (a) control group and (b) Al3+-treated group. (c) BW245C- treated group and (f) CAY10471-treated group raised the number and structure of neurons in model group, whereas (d) BWA868C-treated group and (e) DK-PGD2- treated group decreased the number and structure of neurons in the Al3+-treated group. Sections were pictured at ×400. (B) The intervention effect of DP on the cAMP level in the Al3+-treated group. Values were mean ± SD of six individual experiments. (n = 6, ##P < 0.01 vs. control group. *P < 0.05 vs. Al-treated group, one-way ANOVA with Dunnett’s multiple comparisons) (C) The picture showed the Ca2+ fluorescence of neurons in (a) control group and (b) model group, (c) BW245C and (f) CAY10471 decreased the Ca2+ fluorescence of neurons in model group. (d) BWA868C and (e) DK-PGD2 increased the Ca2+ fluorescence compared with model group (×400). (D) Summary graph showed the intensity of Ca2+ fluorescence relative levels in DP agonists and antagonists groups against model group. Values were mean ± SD of ten individual experiments (n = 10. ##P < 0.01 compared with control group. **P < 0.01 compared with Al3+-treated group, one-way ANOVA with Dunnett’s multiple comparisons).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4835855&req=5

f7: The effects of the DP agonists and antagonists on primary cultured rat hippocampal neurons.(A)The picture showed the number and structure of neurons in (a) control group and (b) Al3+-treated group. (c) BW245C- treated group and (f) CAY10471-treated group raised the number and structure of neurons in model group, whereas (d) BWA868C-treated group and (e) DK-PGD2- treated group decreased the number and structure of neurons in the Al3+-treated group. Sections were pictured at ×400. (B) The intervention effect of DP on the cAMP level in the Al3+-treated group. Values were mean ± SD of six individual experiments. (n = 6, ##P < 0.01 vs. control group. *P < 0.05 vs. Al-treated group, one-way ANOVA with Dunnett’s multiple comparisons) (C) The picture showed the Ca2+ fluorescence of neurons in (a) control group and (b) model group, (c) BW245C and (f) CAY10471 decreased the Ca2+ fluorescence of neurons in model group. (d) BWA868C and (e) DK-PGD2 increased the Ca2+ fluorescence compared with model group (×400). (D) Summary graph showed the intensity of Ca2+ fluorescence relative levels in DP agonists and antagonists groups against model group. Values were mean ± SD of ten individual experiments (n = 10. ##P < 0.01 compared with control group. **P < 0.01 compared with Al3+-treated group, one-way ANOVA with Dunnett’s multiple comparisons).
Mentions: The structures of primary cultured hippocampal neurons were clear, complete and the protrusions of neurons were interwoven into the meshes in the control group. Compared with the control group, the number of neurons was substantial decreased and the protrusions were degenerated in the Al3+-treated group. Compared with the Al3+-treated group, the number of neurons increased and the structures recovered in BW245C-treated group. Treated with BWA868C, the neuronal injury was further aggravated compared with the Al3+-treated group. Treated with DK-PGD2, the hippocampal neurons almost exhibited karyopyknosis and disruption, whereas the bodies and nucleus of neurons were still clear and karyopyknosis compared with the Al3+-treated group. The neuronal injury in the Al3+-treated group was considerable reduced when treated with CAY10471 (Fig. 7A).

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