| Epilepsy, which is essentially an abnormality of cerebral function resulting from the discharges of some cerebral neuron in over high excitement state. Epilepsy is one of the most common diseases or syndromes of central nervous system. Status Epilepticus (SE) is a neurological emergency with an associated mortality rate of 10%–12%, which many affect the patients'life greatly. In patients with temporal lobe epilepsy (TLE), various experimental epilepsy and SE models, the lesions of neurons representing typical apoptotic features were verified by morphological and biochemical methods. The effect of apoptosis on SE has drawn much attention recently though it is still unclear about the detailed signaling pathways that trigger apoptosis is incomplete.Apoptosis is a fundamental process of cell death that occurs via activation of distinct signaling pathways involving mitochondria, mitochondrial regulatory proteins, and activation of Caspases. Ultimately, cells undergo nuclear chromatin condensation, DNA fragmentation, and formation of apoptotic bodies. Apoptosis occurs through three main pathways: the mitochondrial- dependent pathway; the death receptor dependent pathway and the endoplasmic reticulum (ER) pathway. The more important pathway is the mitochondrial-dependent pathway, which induces the leads to the release of Cytochrome c from mitochondria and activation of the death signal. The Cytochrome c quickly combines with the Apaf-1 and Caspase-9, which forms a complex called apoptosome. The activation of Caspase-9 leads to a series of activating of its downstream Caspases, which finally results in programmed cell death.Bcl-2 family proteins, Caspase family proteins and inhibitor of apoptosis proteins (IAPs) are the principal regulators of mitochondrial- dependent apoptotic pathway. The Bcl-2 family of proteins regulates apoptosis by modulating mitochondrial permeability and the release of Cytochrome c. The Bcl-2 family of proteins includes proapoptotic members such as Bax, Bad, and antiapoptotic members such as Bcl-2, Bcl-xL. Although Bad and Bax are located in the cytosol , but translocate to the mitochondria and form a proapoptotic complex with Bcl-2,and then release Cytochrome c. Proteins opposing these pro-apoptotic proteins are antiapoptosis proteins, The antiapoptotic protein Bcl-2 resides in the outer mitochondrial wall, which inhibits the release of pro-apoptotic molecules to mitochondria by multiple mechanisms, including maintenance of mitochondrial membrane integrity and binding to pro-apoptosis members of the Bcl-2 family. The ratio of Bcl-2/Bax protein has also been suggested to determine cell survival or death. Among the BH3-only class of proteins, Bad was the first identified and the best characterized. Bad is a pro-death member of Bcl-2 family that initiates apoptosis by binding to Bcl-2 and Bcl-xL on the outer mitochondrial membrane, causing the release of Cytochrome c into cytosol. The function of Bad is to bind Bcl-2 and Bcl-xl, and block the antiapoptotic action of these proteins. For example: Bad may also inhibit the ability of Bcl-2 and Bcl-xL from forming channels in the mitochondrial membrane, or bind and stabilize the voltage dependent anion channel (VDAC). Bad is an example of a proapoptotic protein that is regulated by phosphorylation, It is phosphorylated on ser-136 by Akt in the PI3K/Akt pathway, Phosphorylation at the site promotes the binding of Bad to 14-3-3 proteins. When bound to 14-3-3 proteins, Bad is sequestered in the cytosol and unable to interact with Bcl-2 or Bcl-xL at the outer mitochondrial membrane. Caspase-9, as well as the other Caspases, exists in an inactive zymogen pro-Caspase state that is activated through proteolytic processing. Cytochrome c, once released in cytosol, interacts with Apaf-1, leading to the activation of Caspase-9 proenzymes. Active Caspase-9 then activates Caspase-3, which subsequently activates the rest of the Caspase cascade and leads to apoptosis. The IAPs are a family of intracellular anti-apoptotic proteins, first identified in Baculovirus, which play a key role in cell survival by modulating death-signaling pathways at a post-mitochondrial level. They currently include X-linked IAP (XIAP), human IAP-1 (Hiap-1), Survivin, Livin and so on. Xchromosome-linked inhibitor of apoptosis protein (XIAP), one of the first mammalian IAP family members cloned, is the most potent inhibitor of apoptosis in IAP family members. It has been shown that XIAP is a direct inhibitor of Caspase-3 and Caspase-9 and can modulate the Bax/Cytochrome c pathway by inhibiting Caspase-9.PI3K (phosphatidylinositol 3-kinase) is a kinase that plays a critical role in signaling pathways important to cell survival. PI3K/Akt signaling pathway is utilized by many cell types for regulating cell cycle, growth, metabolism and inhibition of apoptosis. Akt is known to mediate cell survival by regulating several effectors including Bad, Bcl-2, Bax, Caspase-9, XIAP and so on, which are the important regulatory factors of the mitochondrial-dependent apoptotic pathway meanwhile. The first anti-apoptotic Akt target identified was the pro-apoptotic protein Bad. Activated Akt can phosphorylate Bad specifically at Ser136 and thus can inactivate Bad. Phosphorylated Bad prevents apoptosis, because unphosphorylated Bad can form heterodimers with the anti-apoptotic proteins Bcl-xl or Bcl-2, and then antagonize their anti-apoptotic functions. In addition, researches have also shown that Bax is regulated by phosphorylation of Ser184 in an Akt-dependent manner and that phosphorylation inhibits Bax effects on the mitochondria by maintaining the protein level in the cytoplasm, heterodimerized with antiapoptotic Bcl-2 family members. Akt also phosphorylates and activates the cyclic AMP-response element-binding protein (CREB) on Ser-133, which increases the transcription of anti-apoptotic genes, such as Bcl-2. Akt also phosphorylates the IKK on Thr-23 and releases NF-кB, and then NF-кB up-regulates the expression of Bcl-2. Akt phosphorylates XIAP at residue serine 87 in vitro and in vivo, and interacts with XIAP at physiological protein concentration. These evidences show that Akt and its downstream targets constitute a major cell survival pathway.Erythropoietin (EPO) was originally described for its role in hematopoiesis, which can increase red blood cells by protecting erythroid progenitors against apoptosis. We now know that there are EPO and its receptor (EPO-R) on many cell types of central nervous system in rodent and human brain, such as astrocytes, oligodendrocytes, microglia, and endothelial cells. Exogenous administration of EPO revealed neuroprotective activity in vitro and in vivo in models of central and peripheral neuronal injury occurred in the contexts of trauma, stroke and inflammation. In addition, studies in rodent models of TLE revealed significant effects of rHuEPO(recombinant Human erythropoietin, rHuEPO) in antagonizing the development of SE, but did not determine whether rHuEPO was neuroprotective. Neuroprotective effect induced by rHuEPO applied systemically requires that it crosses the blood–brain barrier (BBB), and many researches have showed rHuEPO administered peripherally crosses the BBB via a receptor-mediated mechanism. In addition, many researches show that pretreatment or even post-treatment of rHuEPO provides protection to neurons after SE. EPO binds to EPO receptor first and then activates janus tyrosine kinase 2 (JAK2), which triggers at least three intracellular signaling cascades through the anti-apoptotic role played by EPO: signal transducer and activator of transcription 5 (STAT5); phosphatidylinositol-3 kinase (PI3K)/Akt and RAS/mitogen-activated protein kinase (MAPK). Pro-survival actions of PI3K occur through activation of the anti-apoptotic effector Akt. Akt activation leads to phosphorylation of certain proteins that promote cell survival. Akt, also known as protein kinase B, is the most well characterized target of PI3K, whose catastaltica would inhibit the activation of Akt. Many experiments presumed the activation of PI3K/Akt signaling pathway through detecting the expression of p-Akt, which showed neuroprotective effects and confirmed by many researches. however, documents demonstrated that EPO could also play neuroprotective effect through the signal pathway of JAK/ STAT; ERK 1/2; NF-κB. To clarify whether the PI3K/Akt signaling pathway involved in the neuroprotective progression originated by rHuEPO in vivo model, we administered LY294002 before rHuEPO treatment, an inhibitor of PI3K. Based on the above mentioned facts, the purpose of our research is to evaluate the anti-apoptosis neuroprotective effect of exogenous rHuEPO post-treatment in PTZ kindled SE rats, and discuss whether rHuEPO modulates the regulatory factors of mitochondrial-dependent apoptotic pathway to mediate the anti-apoptosis neuron protective effects through PI3K/Akt signaling pathway.Part I Changes of p-Akt expression levels and neurons apoptosis in hippocampus of statural epilepticus rats kindled by PTZ and the effect of rHuEPOObjective: To establish a status epilepticus rat model kindled by PTZ and observe the changes of p-Akt expression levels and neurons apoptosis in hippocampus of status epilepticus rat and the effect of rHuEPO.Methods: Adult male Sprague–Dawley (SD) rats were selected to establish a status epilepticus rat model kindled by PTZ. First, a dose of 20 mg/kg was intraperitoneal injection (i.p.), then followed additional doses of 10mg/kg were administered to the rat every 10 min until the onset of SE. Rats were regarded as fully kindled when they exhibited seizure stages 4-5, this procedure lasted for at least 30 minutes. Rats were divided into 5 groups randomly. In the PTZ group, rats were subjected to PTZ i.p.; in the rHuEPO group, rats were subjected to rHuEPO 5000U/kg i.p., the procedure were performed 30 minutes after SE kindled by the PTZ; in LY294002 group, rats were subjected to intracerebral ventricle (ICV) infusion of LY294002 5μl and rHuEPO 5000U/kg i.p., and the procedure were performed 10 min and 30min respectively after SE kindled by PTZ; in DMSO Control group, rats were subjected to ICV infusion of DMSO5μl and rHuEPO 5 000U/kgi.p., and the procedure were performed 10 min and 30min respectively after the SE kindled by the PTZ; in Control group, animals received the same volume of physiological saline i.p. as their paired PTZ- exposed congeners. Then those rats were recorded with electroencephalogram (EEG). 10 rats in each group were anesthetized by 10% chloral hydrate and perfused with 4% paraformaldehyde solution. The brain tissue containing hippocampus was cut and embedded with paraffin. The apoptotic neurons of the hippocampal region were detect through TUNEL method and the positive cell numbers of p-Akt were detected through immunohistochemical SP methods. 10 rats in each group were anesthetized by 10% chloral hydrate and the hippocampi were quickly separated. Total protein for p-Akt was extracted. The expression levels of p-Akt protein were detected by Western blot.Results: EEG changes: the EEG records of the rats in Control group were dominant withαwaves andβwaves and the background was normal without any discharges. The rats in PTZ group were recorded in EEG as sharps, spike-slow waves and most polyspike discharge, with high potential on a relatively normal background. The EEG records of the rats in rHuEPO group, LY294002 group and DMSO group were demonstrated the repressed discharges. The results of TUNEL stains: compared with the Control group, a significant increases for the number of TUNEL-positive cells were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group (P <0.05). Meanwhile, the same increase were also found for the number of TUNEL-positive cells in the PTZ group compared with those in the rHuEPO group, the LY294002 group and the DMSO group (P <0.05). However, compared with the LY294002 group, a significant decrease for the number of TUNEL-positive cells were observed in the rHuEPO group and the DMSO group (P<0.05). The number of TUNEL-positive cells in the DMSO group were higher than in the rHuEPO group, but with no significant difference (P>0.05). The results of immunohistochemical stains: compared with the Control group, the significant increases for the number of p-Akt-IR (immunohistochemical reaction) cells were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group( P<0.05). Meanwhile, the same increases were also found for the number of p-Akt-IR cells in the rHuEPO group, the LY294002 group and the DMSO group (P<0.05) compared with those in the PTZ group. However, there were the significant decreases in the number of p-Akt-IR cells in the LY294002 group compared with those in the rHuEPO group and the DMSO group (P<0.05). The numbers of p-Akt-IR cells in the rHuEPO group were higher than in the DMSO group, but the difference was insignificant (P>0.05). The results of Western blot: the protein levels of p-Akt in hippocampus of rats in each group were determined by p-Akt/β-actin. compared with the Control group, the significant increases for the protein levels of p-Akt were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group (P<0.05). At the same time, the same increase were also found for the protein levels of P-Akt in the rHuEPO group, the LY294002 group and the DMSO group (P<0.05) compared with those in the PTZ group. However, there were significant decrease for the protein levels of P-Akt in the LY294002 group compared with those in the rHuEPO group and the DMSO group (P<0.05). The protein levels of p-Akt in the rHuEPO group were higher than those in the DMSO group, but the difference were insignificant (P>0.05).Conclusion: The status epilepticus rat model kindled by PTZ in the experiment did have impairments of neuronal apoptosis. This model is the perfect animal model which is feasible to study the damaged neuronal apoptosis after status epilepticus. The rHuEPO showed the effect of antiapoptosis neuroprotection in SE rats, the pathway of PI3K/Akt may be one of the neuroprotective ways of rHuEPO. The possible mechanism is rHuEPO activated the PI3K/Akt pathway and then up-regulated the expression of p-Akt to displayed neuroprotective effect.Part II Changes of the Bcl-2 proteins family expression levels in hippocampus of statural epilepticus rats kindled by PTZ and the effect of rHuEPOObjective: To observe the changes of Bcl-2 proteins family expression levels in hippocampus of status epilepticus rat kindled by PTZ and the effect of rHuEPO, and to explore whether the rHuEPO regulated the expression of Bcl-2,Bax,Bad that are the regulatory factors of mitochondrial-dependent apoptotic pathway through PI3K/Akt signaling pathway, and played anti-apoptotic neuroprotective effect.Methods: 10 rats in each group were anesthetized by 10% chloral hydrate and perfused with 4% paraformaldehyde solution. The brain tissue containing hippocampus was cut and embedded with paraffin. the immunohistochemical reaction positive cell numbers of Bcl-2,Bax,Bad were detected through immunohistochemical SP methods. 10 rats in each group were anesthetized by 10% chloral hydrate and the hippocampi were quickly separated. Total RNA was extracted strictly by Trizol. The expression of Bcl-2mRNA,BaxmRNA,BadmRNA was measured by RT-PCR.10 rats in each group were anesthetized by 10% chloral hydrate and the hippocampi were quickly separated. Total protein for Bcl-2,Bax,Bad was extracted. The expression levels of Bcl-2,Bax,Bad protein were detected by Western blot.Results: The results of immunohistochemical stains: compared with the Control group, the significant increases for the number of Bcl-2-IR cells were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group( P<0.05). Meanwhile, the same increases were also found for the number of Bcl-2-IR cells in the rHuEPO group, the LY294002 group and the DMSO group (P<0.05) compared with those in the PTZ group. However, there were the significant decreases in the number of Bcl-2-IR cells in the LY294002 group compared with those in the rHuEPO group and the DMSO group (P<0.05). The numbers of Bcl-2-IR cells in the rHuEPO group were higher than in the DMSO group, but the difference was insignificant (P>0.05). Compared with the Control group, the significant increases for the number of Bax-IR,Bad-IR cells were observed in the PTZ group(P<0.05), the rHuEPO group, the LY294002 group and the DMSO group. Meanwhile, the same increase were also found for the number of Bax-IR,Bad-IR cells in the PTZ group compared with those in the rHuEPO group, the LY294002 group and the DMSO group(P<0.05). However, compared with the LY294002 group, a significant decrease in the number of Bax-IR,Bad-IR cells were found in the rHuEPO group and the DMSO group(P<0.05). The number of Bax-IR,Bad-IR cells in the rHuEPO group were lower than in the DMSO group, but the difference was insignificant (P>0.05).The results of RT-PCR: the expression levels of Bcl-2mRNA,BaxmRNA,BadmRNA were determined by calculating the density ratio of Bcl-2mRNA/β-actin mRNA,BaxmRNA/β-actinmRNA,BadmRNA/β-actinmRNA. Compared with the Control group, the significant increases for Bcl-2mRNA were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group (P<0.05). Meanwhile, the same increase were also found for Bcl-2mRNA in the rHuEPO group, the LY294002 group and the DMSO group compared with those in the PTZ group (P<0.05). However, there were significant decrease for Bcl-2mRNA in the LY294002 group compared with those in the rHuEPO group and the DMSO group (P<0.05). The expression level of Bcl-2mRNA in the rHuEPO group were higher than in the DMSO group, but the difference was insignificant (P>0.05). Compared with the Control group, significant increases for BaxmRNA,BadmRNA were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group(P<0.05). Meanwhile, the same increase were also found for BaxmRNA,BadmRNA in the PTZ group compared with those in the rHuEPO group, the LY294002 group and the DMSO group(P<0.05). However, compared with the LY294002 group, a significant decrease for BaxmRNA,BadmRNA were found in the rHuEPO group and the DMSO group(P<0.05). The expression level of BaxmRNA,BadmRNA in the rHuEPO group were lower than in the DMSO group, but the difference was insignificant(P>0.05).The results of Western blot: The protein levels of Bcl-2,Bax,Bad in hippocampus of rats in each group were determined by Bcl-2/β-actin,Bax/β-actin,Bad/β-actin respectively. compared with the Control group, the significant increases for the protein levels of Bcl-2 were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group (P<0.05). At the same time, the same increase were also found for the protein levels of Bcl-2 in the rHuEPO group, the LY294002 group and the DMSO group (P<0.05)compared with those in the PTZ group. However, there were significant decrease for the protein levels of Bcl-2 in the LY294002 group compared with those in the rHuEPO group and the DMSO group( P<0.05). The protein levels of Bcl-2 in the rHuEPO group were higher than those in the DMSO group, but the difference was insignificant (P>0.05).Compared with the Control group, the significant increase for the protein levels of Bax and Bad were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group(P<0.05). Meanwhile, the same increase were also found for the protein levels of Bax and Bad in the PTZ group compared with those in the the rHuEPO group, the LY294002 group and the DMSO group (P<0.05). However, compared with the LY294002 group, the significant decrease for the protein levels of Bax and Bad were found in the rHuEPO group and the DMSO group (P<0.05). The protein levels of Bax and Bad in the rHuEPO group were lower than those in the DMSO group, but the difference was insignificant (P>0.05).Conclusion: There were elevation in expression levels of Bcl-2,Bax and Bad protein and Bcl-2mRNA,BaxmRNA and BadmRNA in the hippocampus of status epilepticus rat kindled by PTZ, which suggests that Bcl-2,Bax and Bad might participate in the mechanism of hippocampus neuronal apoptosis lesion after status epilepticus. rHuEPO posttreament could increase the expression levels of Bcl-2,Bcl-2mRNA and decrease the expression levels of Bax,BaxmRNA,Bad,BadmRNA, the possible mechanism is the rHuEPO regulated the expression of Bcl-2,Bax,Bad that are the regulatory factors of mitochondrial-dependent apoptotic pathway through PI3K/Akt signaling pathway, and played anti-apoptotic neuroprotective effect.Part III Changes of the Caspase-9 expression levels in hippocampus of statural epilepticus rats kindled by PTZ and the effect of rHuEPOObjective: To observe the changes of Caspase-9 expression levels in hippocampus of status epilepticus rat kindled by PTZ and the effect of rHuEPO, and to explore whether the rHuEPO regulated the expression of Caspase-9 that are the regulatory factors of mitochondrial-dependent apoptotic pathway through PI3K/Akt signaling pathway, and played anti-apoptotic neuroprotective effect.Methods: 10 rats in each group were anesthetized by 10% chloral hydrate and perfused with 4% paraformaldehyde solution. The brain tissue containing hippocampus was cut and embedded with paraffin, the immunohistochemical reaction positive cell numbers of Caspase-9 were detected through immunohistochemical SP methods. 10 rats in each group were anesthetized by 10% chloral hydrate and the hippocampi were quickly separated. Total RNA was extracted strictly by Trizol. The expression of Caspase-9mRNA were measured by RT-PCR.10 rats in each group were anesthetized by 10% chloral hydrate and the hippocampi were quickly separated. Total protein for Caspase-9 was extracted. The expression levels of Caspase-9 protein were detected by Western blot.Results: The results of immunohistochemical stains: compared with the Control group, the significant increases for the number of Caspase-9-IR cells were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group ( P<0.05), Meanwhile, the same increase were also found for the number of Caspase-9-IR cells in the PTZ group compared with those in the rHuEPO group, the LY294002 group and the DMSO group(P<0.05). However, compared with the LY294002 group, a significant decrease in the number of Caspase-9-IR were found in the rHuEPO group and the DMSO group(P<0.05). The number of Caspase-9-IR cells in the rHuEPO group were lower than in the DMSO group, but the difference was insignificant (P>0.05).The results of RT-PCR: the expression levels of Caspase-9mRNA were determined by calculating the density ratio of Caspase-9mRNA/β-actin mRNA. Compared with the Control group, the significant increases for Caspase-9mRNA were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group (P<0.05). Meanwhile, the same increase were also found for Caspase-9mRNA in the PTZ group compared with those in the rHuEPO group, the LY294002 group and the DMSO group (P<0.05). However, compared with the LY294002 group, a significant decrease for Caspase-9mRNA were found in the rHuEPO group and the DMSO group(P<0.05). The expression level of Caspase-9mRNA in the rHuEPO group were lower than in the DMSO group, but the difference was insignificant(P>0.05).The results of Western blot: The protein levels of Caspase-9 in hippocampus of rats in each group were determined by Caspase-9/β-actin. Compared with the Control group, the significant increase for the protein levels of Caspase-9 were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group(P<0.05). Meanwhile, the same increase were also found for the protein levels of Caspase-9 in the PTZ group compared with those in the the rHuEPO group, the LY294002 group and the DMSO group (P<0.05). However, compared with the LY294002 group, the significant decrease for the protein levels of Caspase-9 were found in the rHuEPO group and the DMSO group (P<0.05). The protein levels of Caspase-9 in the rHuEPO group were lower than those in the DMSO group, but the difference was insignificant (P>0.05).Conclusion: There were elevation in expression levels of Caspase-9 protein and Caspase-9mRNA in the hippocampus of status epilepticus rat kindled by PTZ, which suggests that Caspase-9 might participate in the mechanism of hippocampus neuronal apoptosis lesion after status epilepticus. rHuEPO posttreament could increase the expression levels of Caspase-9,Caspase-9mRNA, the possible mechanism is the rHuEPO regulated the expression of Caspase-9 that are the regulatory factors of mitochondrial-dependent apoptotic pathway through PI3K/Akt signaling pathway, and played anti-apoptotic neuroprotective effect.Part IV Changes of the XIAP expression levels in hippocampus of statural epilepticus rats kindled by PTZ and the effect of rHuEPOObjective: To observe the changes of XIAP expression levels in hippocampus of status epilepticus rat kindled by PTZ and the effect of rHuEPO, and to explore whether the rHuEPO regulated the expression of XIAP that are the regulatory factors of mitochondrial-dependent apoptotic pathway through PI3K/Akt signaling pathway, and played anti-apoptotic neuroprotective effect.Methods: 10 rats in each group were anesthetized by 10% chloral hydrate and perfused with 4% paraformaldehyde solution. The brain tissue containing hippocampus was cut and embedded with paraffin. the immunohistochemical reaction positive cell numbers of XIAP were detected through immunohistochemical SP methods. 10 rats in each group were anesthetized by 10% chloral hydrate and the hippocampi were quickly separated. Total RNA was extracted strictly by Trizol. The expression of XIAPmRNA was measured by RT-PCR.10 rats in each group were anesthetized by 10% chloral hydrate and the hippocampi were quickly separated. Total protein for XIAP were extracted. The expression levels of XIAP protein were detected by Western blot.Results: The results of immunohistochemical stains: compared with the Control group, the significant increases for the number of XIAP-IR cells were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group( P<0.05). Meanwhile, the same increases were also found for the number of XIAP-IR cells in the rHuEPO group, the LY294002 group and the DMSO group (P<0.05) compared with those in the PTZ group. However, there were the significant decreases in the number of XIAP-IR cells in the LY294002 group compared with those in the rHuEPO group and the DMSO group (P<0.05). The numbers of XIAP-IR cells in the rHuEPO group were higher than in the DMSO group, but the difference was insignificant (P>0.05). The results of RT-PCR: the expression levels of XIAPmRNA were determined by calculating the density ratio of XIAPmRNA/β-actin mRNA. Compared with the Control group, the significant increases for XIAPmRNA were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group (P<0.05). Meanwhile, the same increase were also found for XIAPmRNA in the rHuEPO group, the LY294002 group and the DMSO group compared with those in the PTZ group (P<0.05). However, there were significant decrease for XIAPmRNA in the LY294002 group compared with those in the rHuEPO group and the DMSO group (P<0.05). The expression level of XIAPmRNA in the rHuEPO group were higher than in the DMSO group, but the difference was insignificant (P>0.05). The results of Western blot: The protein levels of XIAP in hippocampus of rats in each group were determined by XIAP/β-actin. compared with the Control group, the significant increases for the protein levels of XIAP were observed in the PTZ group, the rHuEPO group, the LY294002 group and the DMSO group (P<0.05). At the same time, the same increase were also found for the protein levels of XIAP in the rHuEPO group, the LY294002 group and the DMSO group (P<0.05)compared with those in the PTZ group. However, there were significant decrease for the protein levels of XIAP in the LY294002 group compared with those in the rHuEPO group and the DMSO group( P<0.05). The protein levels of XIAP in the rHuEPO group were higher than those in the DMSO group, but the difference was insignificant (P>0.05).Conclusion: There were elevation in expression levels of XIAP protein and XIAPmRNA in the hippocampus of status epilepticus rat kindled by PTZ, which suggests that XIAP might participate in the mechanism of hippocampus neuronal apoptosis lesion after status epilepticus. rHuEPO posttreament could increase the expression levels of XIAP,XIAPmRNA, the possible mechanism is the rHuEPO regulated the expression of XIAP that are the regulatory factors of mitochondrial-dependent apoptotic pathway through PI3K/Akt signaling pathway, and played anti-apoptotic neuroprotective effect.
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