Characterization of matrix metalloproteinase-2 and-9 in cerebrospinal fluid of clinically normal dogs
Characterization of matrix metalloproteinase-2 and-9 in cerebrospinal fluid of clinically normal dogs
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DOI：
10.2460/ajvr.2002.63.1359
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发表时间：
2002-10-01
影响因子：
1
通讯作者：
Inzana, TJ
中科院分区：
文献类型：
--
作者：
Bergman, RL;Inzana, KD;Inzana, TJ
方法：对23只犬进行脑脊液（CSF）分析，并对犬进行麻醉，收集CSF样品，然后对犬实施安乐死。立即评价每份CSF样本的RBC计数、WBC计数、蛋白质和葡萄糖浓度，并进行细胞学检查。当蛋白质浓度< 25 mg/dL且CSF含有<6 WBC/穆尔和< 25 RBC/穆尔时，认为样品正常。样品储存在-70 ℃。收集脑组织切片并进行处理用于组织学检查。结果-CSF样品的平均WBC计数< 1 WBC/穆尔（范围，0至3 WBC/mL）。平均蛋白浓度为12 mg/dL（范围：8 - 17 mg/dL）。RBC计数为3.65 RBC/穆尔（范围为0至21 RBC/穆尔）。所有CSF样本均对酶谱凝胶产生了明确的禁令，其对应于酶原MMP-2的人商业标准。其他主要的清晰条带在酶谱凝胶上未检测到。在任何样品中均未检测到与MMP-9相关的条带。ELISA结果显示平均+/- SD酶原MMP-2浓度为5.61 +/- 1.92 ng/mL（范围为3.36 - 10.83 ng/mL）。结论和临床相关性-在临床正常犬的CSF中可检测到MMP-2的酶原形式，而MMP-9则不可检测。需要对患有各种神经系统疾病的犬CSF中的MMPs进行进一步研究。
Objective-To characterize matrix metalloproteinase (MMP)-2 and -9 in CSF of clinically normal dogs.Sample Population-Samples of CSF collected from 23 dogs.Procedure-Dogs were anesthetized, CSF samples were collected, and dogs were then euthanatized. Each CSF sample was evaluated immediately for RBC count, WBC count, and protein and glucose concentrations, and cytologic examination also was performed. Samples were considered normal when protein concentration was < 25 mg/dL and CSF contained < 6WBCs/muL and < 25 RBCs/muL. Samples were stored at -70degreesC. Sections of brain tissue were collected and processed for histologic examination. The MMPs were evaluated by use of gelatin zymography and a polyclonal antibody-based sandwich ELISA.Results-Mean WBC count for CSF samples was < 1 WBC/muL (range, 0 to 3 WBCs/mL). Mean protein concentration was 12 mg/dL (range, 8 to 17 mg/dL). an RBC count was 3.65 RBCs/muL (range, 0 to 21 RBCs/muL). All CSF samples generated a clear ban on zymography gels that corresponded to the human commercial standard of proenzyme MMP-2. Other major clear bands were not detected on zymography gels. Bands correlating to MMP-9 were not detected in any samples. The ELISA results revealed a mean +/- SD proenzyme MMP-2 concentration of 5.61 +/- 1.92 ng/mL (range, 3.36 to 10.83 ng/mL).Conclusions and Clinical Relevance-The proenzyme form of MMP-2 is detectable in CSF of clinically normal dogs, whereas MMP-9 is not detectable. Additional investigation of MMPs in CSF from dogs with various diseases of the nervous system is indicated.