The Clinical Value Of Matrix Metalloproteinases-9 For Diagnosing Relapse Status Of Multiple Sclerosis

1. Objective:Multiple sclerosis is a kind of central nervous system(CNS) chronic inflammatory demyelinating diseases.The typical pathological features is that demyelination of CNS,glial cell proliferation and varying degrees of axonal damage. At present a lot of studies have shown that patients with Multiple sclerosis exist blood-brain barrier damage at the early time, and speculate that it may be a critical component of the pathogenesis of Multiple sclerosis.Matrix metalloproteinases-9 is a kind of zinc ion dependent proteases, it can destroy BBB, disintegrate matrix proteins which is in the vascular endothelial cells and myelin basic protein, and then lead to an increased microvascular endothelial permeability, so that T lymphocytes and other inflammatory cells penetrate in CNS through the impaired BBB basement membrane.The typical pathological features of MS,such as the damage of BBB, infiltration of inflammatory cell,degradation of myelin sheath and impairment of axis cylinder are all relate to the abnormally increased activity of MMP-9.The incidence rate of cervical spondylotic radiculopathy is 50-60%, the outstanding of cervical vertebrae intervertebral disk and hyperplasia of links stimulate and oppress cervical nerve root, which lead to cervicobrachialgia commonly in the beginning, and then this kind of pain radiate toward epipodite, the patients will feel paresthesia such as numbness and allergy. These clinical manifestation of cervical spondylotic radiculopathy are similar to MS.The damage of BBB play an important role in the pathophysiological change of MS, the change of microvascular endothelial permeability take place earlier than the characteristic change of MS iconography. Active MS focus can be discovered to be obvious in magnetic resonance imaging(MRI) contrast enhancement, it results frominflammatory change around the pathological blood vessel, increased microvascular endothelial permeability and extravasation of contrast media. MMP-9 is the important component leading to damage of BBB, the serum level of MMP-9 in MS increase suddenly is relate to the new form of focus, so that we can speculate that the level of MMP-9 is different between the relapse and remission status of MS.In our study, We detect the serum levels of MMP-9(matrix metalloproteinases-9)to observe whether MMP-9 detection is the method for differentiating R-R MS(relapse– remission type of Multiple sclerosis)from cervical spondylosis, and to evaluate the clinical value of MMP-9 for differentiating relapse status of MS from remission status of MS.2. Methods:2.1 The group of the experimentThere are five groups in this study: two MS(Multiple sclerosis) groups, two cervical spondylosis groups, and one control group; twenty six patients in relapse status of MS and remission status of MS, thirty patients of cervical spondylosis, and thirty objects without disease. We definite these diseases through CT(computed tomography),EMG(electromyography)and MRI(magnetic resonance imaging) contrast enhancement. We take 2ml venous blood from every people respectively for detection of MMP-9.2.2 The equipments of the experimentThe equipments for detection are ELIASA(enzyme-linked immunosorbent assay)plate,digital display electrothermal incubator with a constant temperature,hydroextractor, pipette, whirlpool admixer, drying oven, tritorium and microdosis motor-operated tissue homogenizer, etc. We use double antibody sandwich ABC-ELISA in this study.First, we take the single antibody of anti-human MMP-9 cover ELIASA plate and let MMP-9 of proof sample and sample combine with the single antibody, then add the biotinylated anti-human MMP-9 in order to form the immune compound,which link with the plate,then make horseradish peroxidase marked Streptavidin combine with biotin,add substrate fluid until it change into blue, in the end add stop buffer- sulphuricacid. We measure the OD figure at the point of 450 nm, because the density of MMP-9is in direct proportion to the OD figure, we can obtain the density of MMP-9 through rendering standard curve, then we analyse the data with statistics.3.Results:3.1 The serum levels of MMP-9 in relapse status and remission status group of MS is higher than the control group respectively, there is statistically significant(P<0.05).3.2 The serum levels of MMP-9 in relapse status and remission status group of cervical spondylosis is same with the control group respectively, there is not statistically significant(P<0.05).3.3 The serum levels of MMP-9 in relapse status and remission status group of MS is higher than cervical spondylosis respectively, there is statistically significant(P <0.05).4. Conclusions:4.1 The serum levels of MMP-9 has the clinical value for differentiating R-R MS(relapse – remission type of Multiple sclerosis)from cervical spondylosis.4.2 To detect the serum levels of MMP-9 regularly in the R-R MS patients can diagnose the relapse of MS in early time, and then prevent the status from deterioration,it can afford help for diagnosis and treatment.4.3 The serum levels of MMP-9 has no clinical value for diagnosing cervical spondylosis.