c-Mpl and TPO expression in the human central nervous system neurons inhibits neuronal apoptosis

Figures

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  Figure 1. Expression of thrombopoietin (TPO) mRNA in human cerebral hemisphere and cerebellum. There was no correlation between the TPO levels in blood plasma and cerebrospinal fluid (CSF). (A) TPO mRNA expression in human cerebral hemisphere and cerebellum was detected by RT-PCR, n = 3. (B) Scatter plot of TPO levels in blood plasma and CSF, r = –0.09, P = 0.80, n = 10.

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  Figure 2. TPO levels in patients with acute cerebral infarction were higher than those in normal people. TPO levels in patients (n = 16) and normal people (control group, n = 45) were detected by ELISA. ** P < 0.01.

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  Figure 3. c-Mpl mRNA and protein were expressed in neural cells and tissues. (A) c-Mpl mRNA expression in human cerebral hemisphere, cerebellum, and C17.2 cells was detected by RT-PCR, n = 3. (B) c-Mpl protein expression in human cerebral hemispheres, hippocampus, cerebellum, brain stem, and spinal cord was detected by immunohistochemistry.

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  Figure 4. TPO demonstrated a neuroprotective effect in a neonatal hypoxic-ischemic rat model. Brain injury was estimated using the percentage of the weight reduction in the ipsilateral cerebral hemisphere compared to the contralateral hemisphere. Brain weights of ipsilateral cerebral hemisphere and contralateral hemisphere were measured in the sham-operated, vehicle-treated, and TPO-treated rats at 1 and 3 weeks of hypoxia-ischemia, n = 16. ** P < 0.01.

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  Figure 5. Brain morphology, histology, NSE staining of cortical neurons, and postural reflex test in sham-operated, vehicle-treated, and TPO-treated pups 3 weeks after surgery. Brain morphology and histology were examined at 3 weeks after surgery. Cortical neurons in the ipsilateral hemispheres were detected using NSE staining. (A) Normal external brain morphology and histology of sham-operated pups. (B) Brain morphology and histology of vehicle-treated pups. (C) Brain morphology and histology of TPO-treated pups. (D) The numbers of cortical neurons in the ipsilateral hemispheres of sham-operated, vehicle-treated, and TPO-treated pups, n = 3. (E) Postural reflex test scores in sham-operated (n = 12), vehicle-treated (n = 16), and TPO-treated pups (n = 16). * P < 0.05.

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  Figure 6. TPO promoted cell proliferation and activated the PI3K/AKT signal in C17.2 cells. C17.2 cells were treated with TPO for 72 h. These data were expressed as means ± SEM. Cell viability was detected by MTT. (A) Cell viability of C17.2 cells with different TPO concentrations, n = 3. (B) Cell viability of C17.2 cells that were treated with the PI3K inhibitor (LY294002, 50 μM) prior to TPO treatment, n = 8. (C) Levels of phosphorylated AKT (p-AKT) at different time intervals. (D) Levels of AKT and p-AKT in the groups treated with TPO and TPO+LY294002. * P < 0.05, ** P < 0.01.

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  Figure 7. Effect of TPO on Bcl-2 and BAX of C17.2 cells at different time intervals. Cells were stimulated for the selected times with 100 ng/mL of TPO. Bcl-2 and BAX were detected by Western blot. (A) Levels of Bcl-2 and BAX at different time intervals. (B) Semiquantitative evaluation of TPO for antiapoptotic effects via Bcl-2/BAX. * P < 0.05, ** P < 0.01.

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  Figure 8. TPO demonstrated a protective effect in a CoCl2-induced PC12 cell injury protection model. Cell viability was detected by MTT. Cell apoptosis and mitochondrial membrane potential were detected by flow cytometry. (A) Effect of different concentrations of CoCl2 on viability of PC12 cells. Cells were treated with CoCl2 for 24 h, n = 3. (B) Protective effect of different concentrations of TPO on CoCl2-induced (500 μmol/L) PC12 cells. Cells were treated with TPO for 48 h, n = 3. (C) Protective effect of 100 ng/mL of TPO on CoCl2-induced (500 μmol/L) PC12 cells. (D) Protective effect of 100 ng/mL of TPO on mitochondrial membrane potential of CoCl2-induced (500 μmol/L) PC12 cells. ** P < 0.01.