Expression Of GR And Inflammatory Factors In Lymphoid Tissue Of Mouse Facial Nerve Palsy Model Induced By Herpes Simplex Virus Type1

Bell's palsy is an acute idiopathic unilateral complete or incomplete peripheral facial nerve paralysis.It's incidence is accounting for50%to70%of of all facial palsy cases. The reasons of Bell's palsy are not yet clear, more and more researchs demonstrated that Herpes simplex virus type1(HSV-1) is the possible reason of this disease. It confirmed this view that researchers succeeded to bulid the animal models of facial paralysis via induced by HSV-1.Although Bell's palsy is a self-limiting disease, a large number of clinical cases have been suggested that the treatment using Glucocorticoids(GC) effectly could Shorten the course, relieve symptoms and improve the cure rate. However, at the present, treatment of GC to facial paralysis is lack of theoretical basis because of limitations of clinical researchs. Medication timing, dosage and treatment of GC for Bell's palsy is lack of Standardize principles, so there are some defects in the clinical treatment.The main mechanism of Anti-inflammatory effects of GC is immunological suppression. Cellular immunity may play a major role on anti-virus after HSV-1infection. Cells of the acquired immune system, specifically T lymphocytes have been considered playing an important role during a neurological injury or disease. Neural immune system interactions such as T lymphocyte circulation within the central nervous system is prepared to respond to stimulus when needed. Sudies have shown that CD4+T cell lymphocytes are benefit for facial motoneuron survival after facial nerve transaction. CD4+T cells can also secrete various cytokines to playing a pro-inflammatory and anti-inflammatory effects, and it can also promote producing CD8+T cells to further remove the virus. So, the CD4+T cells play a important role in the peripheral nerve disease.CD4+T cell have two classic subsets which are T helper type1(Th1) and Th2cell that secrete pro-inflammatory cytokines, such as interferon-γ(INF-γ), interleukin-2(IL-2) or anti-inflammatory cytokines, such as IL-4, IL-15, respectively. IL-2, an inflammatory cytokine, plays a important role in the growth and differentiation of lymphocytes.The potential role of IL-2in peripheral nervous system are remain controversial. IL-4has broad range of biologic and immunological activities and is a important indicator of Th2effect cells.The pattern of GC palys a role of anti-inflammatory is binding with glucocorticoid Receptor (GR). Our preliminary studies demonstrated that the expression of GR on facial nerve motor nucleus and cerebral facial motor cortex in facial palsy mice via HSV-1infection present a dynamic change with time. This experiment reflected the expression pattern of GR in local tissues. In this paper, we will discuss the expression pattern of GR in spleen of facial palsy mice infected by HSV-1and the expression of inflammatory cytokines. Furthermore, we will make tentative researchs on the regulation of GC on these cytokines. PartⅠGlucocorticoid Receptor Expression in Spleen of Mouse Facial Nerve Palsy Model Induced byHSV-1Objective:To research the expression of glucocorticoid receptor(GR) in spleen of mouse facial nerve palsy model induced by herpes simplex virus type1.Methods:Four-week-old male Balb/c mice were selected and randomly divided into three groups including blank control group, saline control group and experimental group. HSV-1was inoculated into the left auricle of experimental mice by scratching with26-gauge needle, and the same volume of saline was placed in the left auricle of control group. The spleens were taken out at6h,12h,24h,2d,3d,5d and7d after producing facial paralysis in mice.Western blot and immunohistochemistry were used to detect the level of GR protein, Real-time PCR was used to measure the level of GR mRNA.Results:Expression of GR mRNA in spleens of experimental group mice was slightly decreased after producing facial paralysis, sharply declined at24h, showing a significant difference compared with saline control group (p<0.05).The expression gradually reached a peak at3d, having a significant difference compared with saline control group (p<0.05). At7d,The expression eventually returned to the baseline level which had no difference with saline control group(P>0.05).GR protein expression in experimental group was similar to GR mRNA.Conclusion:Expression of GR in spleen of mouse facial nerve palsy model induced by herpes simplex virus type1appears in a time-depend pattern which is first reduce,then increase and finally return to baseline level.It prompts that GR in spleen of the model shows a regularity expression,and it maybe play a role in the process of the disease. Part ⅡInflammatory factors expression in the lymphoid organs of mouse facial nerve palsy model induced by HSV-1Objective:To research the expression of CD4+T lymphocyte,IL-2and IL-4in the mouse dranining cervical lymph node(DCLN) and spleen during_facial nerve palsy model induced by herpes simplex virus type1(HSV-1) and the inhibitory effects of glucocorticoids..Methods:Four-week-old male Balb/c mice were selected and randomly divided into three groups including normal control group(group N), saline control group(group S) and experimental group(group H). HSV-1was inoculated into the surface of posterior auricle of mouse of group H to set up an animal model. The paralyzed mice were divided in three groups as A,B and C. In group A, Mice were sacrificed at1,3d,5d,7d after producing facial paralysis in mice. In other two groups, were injected daily for three days with methylprednisolone sodium succinate (MPSS) or with combined administration of MPSS and glucocorticoid receptor blocker (RU486). The expression and of CD4+T lymphocyte,IL-2and IL-4in the DCLN,MLN and spleen was detected by Fluorescence Activted cell sorter(FACS).Results:In group A, CD4+T lymphocyte in DCLN reached the peak at the5d(39.30±0.98%) after mouse producing facial paralysis, and the peak time was the7d(28.97±1.10%) in spleen. The expressions of CD4+IL-2+and CD4+IL-4+cells in DCLN and spleen reached the peak was5d(1.76±0.21%,1.11±0.13%) and7d(2.16±0.33%,1.54±0.26%) respectively after mouse had developd facial nerve palsy, which were significantly higher than those in the group S and group N(p<0.05). The numbers above-mentioned in the MLN of mouse have no significant difference between group A and S,N. Besides (p>0.05), the expression of the CD4+T lymphocyte,CD4+IL-2+and CD4+IL-4+cells could be inhibited by MPSS, and the datas showed significant differences compared with group A and group C(p<0.05). Conclusion:It prompts that CD4+T lymphocyte,IL-2and IL-4participate in the immune response of the disease,which was take place in DCLN and spleen. MPSS can effectively attenuate HSV-1-mediated damages in nerve system, which is closely associated to its inhibitory effect on expressions of the cells. Background:To research the expression of glucocorticoid receptor(GR) in spleen of mouse facial nerve palsy model induced by herpes simplex virus type1.Methods:Four-week-old male Balb/c mice were selected and randomly divided into three groups including blank control group, saline control group and experimental group.HSV-1was inoculated into the left auricle of experimental mice by scratching with26-gauge needle, and the same volume of saline was placed in the left auricle of control group. The spleens were taken out at6h,12h,24h,2d,3d,5d and7d after producing facial paralysis in mice.Western blot and immunohistochemistry were used to detect the level of GR protein, Real-time PCR was used to measure the level of GR mRNA.Results:Expression of GR mRNA in spleens of experimental group mice was slightly decreased after producing facial paralysis, sharply declined at24h, showing a significant difference compared with saline control group (p<0.05).The expression gradually reached a peak at3d, having a significant difference compared with saline control group (p<0.05). At7d,The expression eventually returned to the baseline level which had no difference with saline control group(p>0.05). GR protein expression in experimental group was similar to GR mRNA.Conclusion:Expression of GR in spleen of mouse facial nerve palsy model induced by herpes simplex virus type1appears in a time-depend pattern which is first reduce,then increase and finally return to baseline level.It prompts that GR in spleen of the model shows a regularity expression,and it maybe play a role in the process of the disease Background:To research the expression of CD4+T lymphocyte,IL-2and IL-4in the mouse dranining cervical lymph node(DCLN) and spleen during_facial nerve palsy model induced by herpes simplex virus type1(HSV-1) and the inhibitory effects of glucocorticoids.Methods:HSV-1was inoculated into the surface of posterior auricle of mouse to set up an animal model. The paralyzed mice were divided in three groups(group A,B,C)as detailed in text. Mice, in group A, were sacrificed at different time points and, in other two groups, were injected daily for three days with methylprednisolone sodium succinate (MPSS) or with combined administration of MPSS and glucocorticoid receptor blocker (RU486). The expression and of CD4+T lymphocyte,CD4+IL-2+and CD4+IL-4+cells in the DCLN and spleen was detected by Fluorescence Activted cell sorter(FACS).Results:In group A, CD4+T lymphocyte in DCLN reached the peak at the5d after mouse producing facial paralysis, and the peak time was the7d in spleen.The expresstions of CD4+IL-2+and CD4+IL-4+cells in DCLN and spleen reached the peak was5d and7d respectively. Besides, the expression of the cells could be inhibited by MPSS, All the datas showed significant differences compared with control group individuals(P<0.05).Conclusion:It prompts that CD4+T lymphocyte,IL-2and IL-4participate in the immune response of facial nerve palsy induced by HSV-1,which was take place in DCLN and spleen. MPSS can effectively attenuate HSV-1-mediated damages in nerve system, which is closely associated to its inhibitory effect on expression of the cells.