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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

Levels of DP1, DP2 and L-PGDS in primary cultured rat hippocampal neurons detected by reverse transcription polymerase chain reaction.(A) The expressions of L-PGDS, DPs mRNA were measured by RT-PCR. The relative mRNA level of L-PGDS, DPs were standardized to endogenous β-actin mRNA for each sample. Al administration caused the significant increase of L-PGDS, DP1 levels and decrease of DP2 level compared with the control group. (B) The expressions of L-PGDS, DPs proteins were measured by WB. The relative protein levels of L-PGDS, DPs were standardized to endogenous β-actin protein for each sample. Al administration caused the significant increase of L-PGDS, DP1 levels and decrease of DP2 level compared with the control group. Values were mean ± SD (n = 3, **P < 0.01 vs. control group, one-way ANOVA with Dunnett’s multiple comparisons).
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f3: Levels of DP1, DP2 and L-PGDS in primary cultured rat hippocampal neurons detected by reverse transcription polymerase chain reaction.(A) The expressions of L-PGDS, DPs mRNA were measured by RT-PCR. The relative mRNA level of L-PGDS, DPs were standardized to endogenous β-actin mRNA for each sample. Al administration caused the significant increase of L-PGDS, DP1 levels and decrease of DP2 level compared with the control group. (B) The expressions of L-PGDS, DPs proteins were measured by WB. The relative protein levels of L-PGDS, DPs were standardized to endogenous β-actin protein for each sample. Al administration caused the significant increase of L-PGDS, DP1 levels and decrease of DP2 level compared with the control group. Values were mean ± SD (n = 3, **P < 0.01 vs. control group, one-way ANOVA with Dunnett’s multiple comparisons).

Mentions: Compared with control group, the expression of L-PGDS and DP1 mRNA up-regulated 100% in the model group which was treated with Al (malt)3 (P < 0.01). By contrast, the expression of DP2 mRNA decreased significantly more than 70% in the model group (P < 0.01) (Fig. 3A).


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)

Levels of DP1, DP2 and L-PGDS in primary cultured rat hippocampal neurons detected by reverse transcription polymerase chain reaction.(A) The expressions of L-PGDS, DPs mRNA were measured by RT-PCR. The relative mRNA level of L-PGDS, DPs were standardized to endogenous β-actin mRNA for each sample. Al administration caused the significant increase of L-PGDS, DP1 levels and decrease of DP2 level compared with the control group. (B) The expressions of L-PGDS, DPs proteins were measured by WB. The relative protein levels of L-PGDS, DPs were standardized to endogenous β-actin protein for each sample. Al administration caused the significant increase of L-PGDS, DP1 levels and decrease of DP2 level compared with the control group. Values were mean ± SD (n = 3, **P < 0.01 vs. control group, 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

f3: Levels of DP1, DP2 and L-PGDS in primary cultured rat hippocampal neurons detected by reverse transcription polymerase chain reaction.(A) The expressions of L-PGDS, DPs mRNA were measured by RT-PCR. The relative mRNA level of L-PGDS, DPs were standardized to endogenous β-actin mRNA for each sample. Al administration caused the significant increase of L-PGDS, DP1 levels and decrease of DP2 level compared with the control group. (B) The expressions of L-PGDS, DPs proteins were measured by WB. The relative protein levels of L-PGDS, DPs were standardized to endogenous β-actin protein for each sample. Al administration caused the significant increase of L-PGDS, DP1 levels and decrease of DP2 level compared with the control group. Values were mean ± SD (n = 3, **P < 0.01 vs. control group, one-way ANOVA with Dunnett’s multiple comparisons).
Mentions: Compared with control group, the expression of L-PGDS and DP1 mRNA up-regulated 100% in the model group which was treated with Al (malt)3 (P < 0.01). By contrast, the expression of DP2 mRNA decreased significantly more than 70% in the model group (P < 0.01) (Fig. 3A).

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