Received:01 June 2016,
Published:15 February 2017
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Zhixin Zhang, Yinglin Cui, Yanjie Li, et al. BAMBI inhibits apoptosis in glutamate induced neurons and astrocytes in spinal dorsal horn of rats
Zhixin Zhang, Yinglin Cui, Yanjie Li, et al. BAMBI inhibits apoptosis in glutamate induced neurons and astrocytes in spinal dorsal horn of rats
Objective
2
To determine the effect of the over-expression of BMP and activin membrane-bound inhibitor (BAMBI) on the apoptosis of rat neurons and astrocytes in spinal dorsal horn induced by glutamate (Glu).
Methods
2
Neurons and astrocytes were isolated from spinal dorsal horn of newborn Wistar rats
and primarily cultured. Then the cells were randomly divided into 6 groups: control group
Glu group
Glu+pcDNA group
Glu+pcDNA-BAMBI group
Glu+siRNA control group and Glu+BAMBI siRNA group. The expression of BAMBI was measured by Western blotting. Cell apoptosis was detected by flow cytometry. The expression of microtubule associated protein light chain 3 (LC3)
Beclin1 and p62 was measured by Western blotting. The content of GFP-LC3 in the cultured cell groups was observed by fluorescence microscopy. Finally
the levels of tumor necrosis factor-α (TNF-α)
interleukin-1β (IL-1β) and cleaved-Caspase3 were detected by Western blot analysis.
Results
2
Compared to the control cells
Glu treatment induced cell apoptosis
decreased the expression of LC3-Ⅰ
promoted the expression levels of LC3-Ⅱ
Beclin1 and p62
increased the content of GFP-LC3
and elevated the level of TNF-α
IL-1β and cleaved-Caspase 3 (
P
<
0.05). However
when compared with Glu treatment group
BAMBI overexpression inhibited the cell apoptosis
strongly promoted the expression of LC3 and Beclin1
while decreased the levels of LC3-Ⅰ and p62
significantly declined the levels of TNF-α
IL-1β and cleaved-Caspase3 (
P
<
0.05)
and induced a large accumulation of GFP-LC3 in the cells. The effects of BAMBI silence group was opposite to those of the overexpression.
Conclusion
2
The overexpression of BAMBI could enhance autophagy and inhibit apoptosis in glutamate-induced neurons and astrocytes in rat spinal dorsal horn.
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