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BACKGROUND AND PURPOSE spinal cord injury (spinal cord injury, SCI) level wartime common a serious nervous system trauma, and often leads to a patient's limbs, varying degrees of paralysis or sphincter disturbances, not only to cause the patient pain, and also increase the burden on the social and family . SCI world total incidence of 20-40 million / person / year, the United States the incidence of 28 to 55 people / million / year, an annual increase of about 1 million cases of injury when the average age was 31.7 years old. Care and treatment of these patients, the total cost of more than seven billion U.S. dollars annually. No comprehensive statistics in China, but Beijing statistical incidence of spinal cord injury in 2002 was 60 / million, much higher than the overall level in the United States and the world, as China's economic development, transportation, construction and coal mine accidents as a result of the SCI Vietnam increasingly become the primary threat to human health and quality of life issues. SCI due to disability is currently no effective treatment for recovery of neurological function has been the clinical treatment of the problem, the research on spinal cord injury and repair in terms of its social significance or scientific sense, are very necessary and urgent. Current research on spinal cord injury has made considerable progress, summarized in the following aspects: (1) neuroprotection (Neuroprotection), in addition to the primary injury after spinal cord trauma, more importantly, through a variety of mechanisms (such as hypoxia-ischemia, excitatory amino acid toxicity, ion homeostasis, calcium overload, apoptosis and inflammation) lead to secondary damage is the main reason for the loss caused by spinal cord function. The primary damage is difficult to save, and secondary injury can rescue. (2) nerve regeneration (Neuroregeneration), research in this area in the 1990s progress, the main findings of the central nervous system has the ability to regenerate, but because the external microenvironment causes inhibition of nerve regeneration, nerve regeneration inhibitory molecule, glial scarring and cysts, and other physical and chemical barriers. (3) nerve graft (Neurotransplantation), mainly through peripheral nerve tissue engineering (various sources) stem cell transplant bridging necrosis areas, promoting regeneration to repair the injured spinal cord. (4) neural plasticity neural remodeling (Neuroplasticity) study found that in patients after SCI can restore some of the features, and spinal cord plasticity spontaneously, as long as the residual small portion of the nerve fibers of the spinal cord function reconstruction that Help, clinical through rehabilitation therapy to help patients with spinal cord function reconstruction. While the above strategy has achieved good results in animal studies, but the clinical efficacy is unsatisfactory, evidence-based medicine evidence to show that the drug for the secondary injury, such as the the high hopes NMDA receptor inhibitor, nerve ganglioside fat, free radical scavenging agents are invalid SCI. Shows that promote spinal cord injury repair and regeneration strategy the road is a long way off, therefore, to find a spinal cord injury and repair a new breakthrough and new therapeutic targets, with important medical value and social significance. Early it has been noted that the central nervous system (central nervous system, CNS) injury, the female patient's functional recovery and prognosis often than men, further clinical studies (including epidemiological survey) and animal experiments have also been similar discovery and promotion of neural trauma and neurodegenerative diseases (such as Alzheimer's disease, Parkinson's disease), many scholars believe that this major and estrogen, and that estrogen has a clear protective effect of nerve damage is. A long time, the soluble cell estrogen receptors of ERα / β (ER) has been considered to be the only receptor of its role. Class receptors by estrogen binding, conformational change, and transferred to the nucleus with specific DNA Series - estrogen response element (estrogen responsive element, ERE) combination, regulation of transcription and expression of a series of genes, which affect cell the development, proliferation, and apoptosis. The classical pathway, this pathway is an estrogen generally takes a long time, the need for several hours to several days, referred to as the slow gene action of estrogen. However, a large number of studies have shown that estrogen in tens of seconds, a few minutes to tens of minutes onset, such as the regulation of vascular tone, and quick to promote the release of NO, Ca2 mobilization, etc.. In addition to the classic slow gene pathways are suggesting that estrogen, yet by not awareness of non-genetic mechanism for rapid onset. Science published an article in 2005 is ample evidence that the existence of the new estrogen receptor-GPR30 (a G protein-coupled receptor)-mediated pathway of estrogen, as the rapid effects of estrogen-mediated membrane receptor in the protective effect of estrogen on the spinal cord injury may play an important role, is an important new target for spinal cord injury repair research. Purpose. Distribution pattern observed GPR30 space in the spinal cord of normal rats and expression in the spinal cord cells. 2. Understand SCI after E2, E2-BSA, G-1, saline motor function in rats after intervention recovery and GPR30 protein expression. Method 1. The subject for the study of adult female SD rats, morphological studies of normal adult rat spinal cord: immunohistochemical staining method to observe GPR30 in the spinal cord of the cervical, thoracic, lumbar, sacral segmental The spatial distribution characteristics; cellular localization using immunofluorescence double labeling was observed GPR30 in the rat spinal cord, and to explore the possible role of GPR30 in spinal cord injury. 2 to establish combat injuries of the spinal cord experimental model, around the normal group, the contrast between the SCI group, SCI Group E2, SCI E2-BSA group, SCG-group 1 for the following studies: 1) functional studies: The rats of each group rat lower limb motor function score (BBB score), compare each set of results, and discussed the role of GPR30 in SCI. 2) the use of Western-blotting technology to interfere with each group of spinal cord tissue GPR30 protein levels were detected compared between GPR30 protein levels, further reveal the role of GPR30 in SCI. Results 1.GPR30 in the normal rat spinal cord segment space distribution: GPR30 can be seen to have a clear expression of the various segments of the rat spinal cord, but the main expression in the spinal cord gray matter, in the white matter of the distribution segment basic similar evenly distributed, no expression intensive, but there are some differences in the gray matter of the distribution. GPR30 mainly distributed in the gray matter of the cervical spinal cord in the ventral horn of the gray matter around the central canal also seen more expression, no GPR30 expression in the dorsal horn of the basic, but GPR30 positive signal is visible near the dorsal horn of the gray matter. GPR30 is still mainly in the thoracic spinal cord gray matter in the ventral horn of distribution, but only see a very small amount of the distribution around the central canal were no obvious GPR30 positive signal in the periaqueductal gray dorsal horn and dorsal horn. GPR30 is still mainly in the lumbar spinal cord gray matter distribution in the ventral horn, visible in the gray matter of joint distribution of a very small amount, but in the dorsal horn and periaqueductal gray seen a lot of GPR30 positive signal distribution. Visible in the sacral intramedullary GPR30 is still mainly in the gray matter of the ventral horn of distribution and the distribution is relatively sparse, and no significant aggregation, no significant distribution in the gray matter of the dorsal horn. From the cell structure is positioned within the the GRAY cell in the spinal cord of each segment GPR30 are expressed in the cytoplasmic membrane, and GPR30 positive signals were not detected in the nucleus. 2.GPR30 expression in normal rat spinal cord cells: use double immunofluorescence staining techniques GPR30 nerves in the spinal cord of normal rat spinal cord staining observed yuan clear expression in oligodendrocytes microglia also expressed, but no expression in astrocytes. Functional studies: BBB score after sham controls after SCI SCI group, E2, E2-BSA group, G-1 group packet given intervention, the results are as follows: 4. Detected by Western-blotting intervention GPR30 protein expression levels groups: saline group, E2 group, E2-BSA group and the G-1 group GPR30 protein levels than those of the normal control group. The injury visible saline group protein expression among the intervention group was significantly lower than the E2 group, E2-BSA group, G-1 group, saline group the protein expression of growth trends are consistent with the E2 group. E2 protein levels increased slowly in the early stages after injury, reached the highest value in the fourteenth day, after which protein expression levels slowly decreased protein expression levels at days 1, 3, E2 group was significantly higher than that of E2-BSA group, but is visible in the first seven days a rapid increase in protein expression levels of E2-BSA group, the expression was significantly more than E2 group. E2-BSA group also increased the level of protein expression within 3 days after injury is very slow, but the expression level of the highest value on the seventh day or so. G-1 protein expression levels is to start early segments after injury a rapid increase in the first three days, have been significantly higher than the other groups, and reached the highest value in about seven days, and thereafter the amount of protein to start fast reduced, and in the first 14 days was significantly lower than the expression level of the E2 group. Conclusions normal adult male rat spinal cord, estrogen membrane receptor GPR30 was expressed in the gray matter of the spinal cord, but is mainly distributed in the gray matter, and expression in motor neurons of the spinal cord ventral foot. Spinal cord cells in normal adult male rats, GPR30 in the cytoplasm of neurons expressed in microglia and oligodendrocytes clear expression in astrocytes no clear expressed. In the present experimental conditions, ordinary estrogen E2, combined with bovine serum albumin of estrogen E2-BSA (impermeable membrane) and GPR30-specific agonist G-1 are of SCI has a clear protective effect, and G-1 in early after SCI protective effect of estrogen than ordinary. 4 estrogen membrane receptor GPR30 participation mediated protective effect of estrogen on the SCI, but other ways of estrogen also may be involved in mediated by such effects.
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