Malibatol A enhances alternative activation of microglia by inhibiting phosphorylation of Mammalian Ste20-like kinase1 in OGD-BV-2 cells.
OBJECTIVE
To investigate the polarization effect of Malibatol A on oxygen-glucose deprivation (OGD)-BV-2 cells, and the possible molecular mechanism involved in c-Abl-MST signaling pathway.


METHODS
The OGD BV-2 cell model was established. BV-2 cells were exposed to OGD for 8 h followed by reperfusion for 15 h with Malibatol A at different concentration of 0.5, 1, 2, 4, 8, 16 μM or without it. And then cells, mRNA and protein were harvested respectively. The cell viability and apoptosis were measured by MTT assay and flow cytometry. The mRNA of classical activated microglia (M1) markers (MCP-1, IL-1 and TNF-α) and alternatively activated microglia (M2) markers (Ym-1, CD206, IL-10, TGF-β) in BV-2 cells were measured by RT-PCR. Meanwhile, the proteins of Ym-1 and CD206 was assayed by flow cytometry. Furthermore, the expression of c-Abl and MST was measured by Western blot.


RESULTS
Malibatol A significantly decreased apoptosis and increased viability of OGD BV-2 cells in a dose-dependent manner. In the presence of Malibatol A, the mRNA levels of Ym-1, CD206, IL-10 and TGF-β mRNA was significantly increased in OGD-BV-2 cells, while the mRNA levels of MCP-1, IL-1 and TNF-α was obviously down-regulated. Meanwhile, the proteins of Ym-1 and CD206 was raised in OGD BV-2 cells with Malibatol A. Besides, Malibatol A also inhibited OGD-induced p-MST1(Y433) in BV-2 cells.


CONCLUSIONS
Malibatol A could attenuate OGD-induced BV-2 cell injury and promote M2 microglia polarization. The mechanism may be related to inhibition of MST1 phosphorylation at Y433.
a Department of Neurology , Drum Tower Hospital, Medical School of Nanjing University , Nanjing , PR China.
Leihua
L
Weng
b Department of Intensive Care Unit , Drum Tower Hospital, Medical School of Nanjing University , Nanjing , PR China.
Zhengzheng
Z
Wu
c Department of Neurology , Affiliated Zhongshan Hospital of Xiamen University , Xiamen , China.
Weihong
W
Zheng
a Department of Neurology , Drum Tower Hospital, Medical School of Nanjing University , Nanjing , PR China.
Hailan
H
Meng
a Department of Neurology , Drum Tower Hospital, Medical School of Nanjing University , Nanjing , PR China.
Lijuan
L
Han
a Department of Neurology , Drum Tower Hospital, Medical School of Nanjing University , Nanjing , PR China.
Sulei
S
Wang
d State Key Laboratory of Brain and Cognitive Sciences , Beijing , China.
Zengqiang
Z
Yuan
a Department of Neurology , Drum Tower Hospital, Medical School of Nanjing University , Nanjing , PR China.|e Jiangsu Key Laboratory for Molecular Medicine , Medical School of Nanjing University , Nanjing , China.|f Diagnosis and Therapy Center of Stroke in Jiangsu Province , Nanjing , China.
Yun
Y
Xu
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002461:Cell Line, Transformed; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000031:analogs & derivatives / Q000378:metabolism; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000172:deficiency*; D000860:Hypoxia / Q000473:pathology*; D037181:Lectins, C-Type / Q000378:metabolism; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins / Q000378:metabolism; D051379:Mice; D017628:Microglia / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013267:Stilbenes / Q000494:pharmacology*; D000099317:Hippo Kinases
D004317:Doxorubicin; D005947:Glucose; D037181:Lectins, C-Type; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D017346:Protein Serine-Threonine Kinases; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D013267:Stilbenes; C111734:malibatol A; C079109:N-(3-pyridylmethyl)adriamycin; C490660:Stk4 protein, mouse; D000099317:Hippo Kinases
MST1; Malibatol A; Oxygen-glucose deprivation; Polarization