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Store-Operated Ca2+ Channels Blockers Inhibit Lipopolysaccharide Induced Astrocyte Activation

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Abstract

The destruction of calcium homeostasis is an important factor leading to neurological diseases. Store-operated Ca2+ (SOC) channels are essential for Ca2+ homeostasis in many cell types. However, whether SOC channels are involved in astrocyte activation induced by lipopolysaccharide (LPS) still remains unknown. In this study, we used LPS as an exogenous stimulation to investigate the role of SOC channels in astrocyte activation. Using calcium imaging technology, we first found that SOC channels blockers, 1-[h-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole (SKF-96365) and 2-aminoethyldiphenyl borate (2-APB), inhibited LPS induced [Ca2+]i increase, which prompted us to speculate that SOC channels may be involved in LPS induced astrocyte activation. Further experiments confirmed our speculation shown as SOC channels blockers inhibited LPS induced astrocyte activation characterized as cell proliferation by MTS and BrdU assay, raise in glial fibrillary acidic protein expression by immunofluorescence and Western Blot and secretion of interleukin 6 (IL-6) and interleukin 1β (IL-1β) by ELISA. So, our studies showed that SOC channels are involved in LPS-induced astrocyte activation.

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Acknowledgments

We would like to thank Karrune Woan and Jun-Min Zhou for assistance in preparation of the Main manuscript. This work was supported by Natural Science Foundation of Guangdong Province (NO. 8151018201000039), Key Research Foundation of Guangzhou Education Bureau (2012C043) to JHL, National Natural Science Foundation of China (NO. 81100986), Research Foundation of Guangzhou Education Bureau (NO. 10A154) and Breeding Project of Guangdong Province (NO. LYM11105) to ZST.

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The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Department of Physiology, Guangzhou Medical University, Guangzhou, People’s Republic of China

    Jian-Hua Li, Shen-Ting Zhao, Miao-Ru Peng & Xiao-Ai Liu

  2. Department of Neurobiology, Southern Medical University, Guangzhou, People’s Republic of China

    Cui-Ying Wu, Xiong Cao, Shu-Ji Li & Tian-Ming Gao

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Jian-Hua Li and Shen-Ting Zhao contributed equally to this work.

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11064_2013_1130_MOESM1_ESM.ppt

Supplemental Fig 1 Increased cell viability by LPS for 48 h. The viability was examined by MTS assay. n=3. Results are mean±SE of different independent experiments. *P < 0.05 vs. control group (PPT 64 kb)

11064_2013_1130_MOESM2_ESM.ppt

Supplemental Fig 2 Increased percentage of BrdU positive cells by LPS for 48 h. Data of BrdU positive cells were presented as percentage of the control group. n=15. Results are mean±SE of different independent experiments. *P < 0.05 vs. control group (PPT 64 kb)

11064_2013_1130_MOESM3_ESM.ppt

Supplemental Fig 3 GFAP increased expression induced by LPS. Cells were incubated with 0.5μg/ml LPS for different incubation time. (a) The immunofluorescence of GFAP (green), Co-staining with DAPI (blue) allowed the identification of the nucleus. (b) Representative western blot showing the expression of GFAP. (c) Quantification of the expression of GFAP. Results are mean±SE of four different independent experiments. *P < 0.05 vs. 0 h group (PPT 6914 kb)

11064_2013_1130_MOESM4_ESM.ppt

Supplemental Fig 4 Increased IL-6 and IL-1β by LPS. Cells were incubated with 0.5μg/ml LPS for different incubation time. (a) The level of IL-6. n=4. (b) The level of IL-1β. n=4. Results are mean±SE of four different independent experiments. *P < 0.05 vs. 0 h group (PPT 274 kb)

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Li, JH., Zhao, ST., Wu, CY. et al. Store-Operated Ca2+ Channels Blockers Inhibit Lipopolysaccharide Induced Astrocyte Activation. Neurochem Res 38, 2216–2226 (2013). https://doi.org/10.1007/s11064-013-1130-0

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