ABSTRACT
Objective To investigate the effects and mechanisms of Glutamine (Gln) supplementation on neurobehavioral outcome,neuronal apoptosis,microglia polarization,and neuroinflammatory response after traumatic brain injury (TBI) in rats.Methods TBI animal models were established using Feeney's method.Sixty Wistar rats were randomly divided into three groups:control group (Con),traumatic brain injury group (TBI),and glutamine supplementation group (TBI+Gln).We measured rat behavioral outcomes by modified neurologic severity score (mNSS) tests at day 1,3,7 and 14 after TBI.Apoptotic neurons were determined by Nissl staining.The microglia polarization relatived protein (Iba-1,CD16,CD86) expressions in TBI cerebral cortices were determined by immunohistochemistry staining and western blotting,respectively.While,the serum levels of tumor necrosis factor-α (TNF-α),interleukin (IL)-1 and interferon (IFN)-γ were tested by enzyme linked immunosorbent Assay (ELISA).Results Compared with the Con group,the levels of neurobehavioral outcome,neurons apoptosis,microglia polarization and neuroinflammatory factors were significantly increased in the other two groups (P=0.00).Compared with the TBl group,glutamin supplementation improvedneurobehavioral outcome [7 d:(10.74±0.25) points vs.(8.94±0.24) points,P=0.01;14 d:(8.77± 0.16) points vs.(7.43±0.13) points,P=0.03].Meanwhile,glutamin supplementation suppressed the apoptotic rates of neurons [3d:(80.18±8.38)％ vs.(65.47±7.02)％,P=0.01;7 d:(58.90±6.12)％ vs.(42.73±4.88)％,P=0.01;14d:(39.56±2.95)％vs.(31.12±3.16)％,P=0.01],inhibited protein expressions of Iba-1 and CD16,and increased the protein expression of CD86,which promoted the phenotypic shift of microglia from M1 to M2 phenotype,inhibited microglial activation,and thus reduced TBI-induced neuroinflammatory factors [TNF-α:(125.42 ± 12.81) pg/ml vs.(74.36 ± 9.25) pg/ml,P =0.01;IL-1:(69.04±8.48) pg/ml vs.(34.73±3.92) pg/ml,P=0.01;TNF-α:(89.75±9.40) pg/ml vs.(45.62±6.64) pg/ml,P=0.02].Conclusion Glutamine supplementation can markedly reduce neuron apoptosis and improve neurological outcomes after TBI,possibly mediated by promoting the phenotypic shift of microglia from M1 to M2 phenotype and thus reducing TBI-induced neuroinflammatory factors.
ABSTRACT
BACKGROUND: Transplantation of fragments of ovarian cortex is performed without vascular reanastomosis, therefore, to increase the tolerance of ovarian tissue to the freezing-thawing and ischemic injuries is critical for follicular survival and functional longevity of the graft.OBJECTIVE: To investigate the effect of follicle stimulating hormone (FSH) on the morphological and function of sheep ovary tissue in the process of cryopreservation, so that to provide new freezing method for human ovary tissues. METHODS: Healthy BALB/c strain female nude mice were equally and randomly divided into 3 groups and subjected to heterotopic transplantation of fragments of sheep ovarian cortex. (1) Control group: transplantation following sampling; (2) experimental group: transplantation following freezing-thawing, and the solution was free of FSH; (3) FSH group: the freezing and thawing and culture fluid contained FSH. The estrous cycle recovering rate, estrous cycle recovering time, the number of follicle were observed following transplantation. The histological changes, and the level of E2 in blood serum were observed at 4 weeks after transplantation.RESULTS AND CONCLUSION: There were no significant differences in estrous cycle recovering rate and number of follicle/PHF between FSH and control groups (P> 0.05), but the number of follicle was greater than the experimental group (P< 0.05). Moreover, the time of estrous cycle recovering in FSH group was similar to control group, but shorter than experimental group (P < 0.05). There were more developing follicular in FSH group, but few in experimental group at 4 weeks. No significant difference was detected in E2 level in blood serum between FSH and control group (P > 0.05), but significantly greater than the experimental group (P< 0.05). Results show that FSH addition in vitrification fluid can improve ovarian follicle survival following cryopreserved sheep ovary tissue transplantation.