Ozkan H, Akkoç N, Aydin A, Olgun N, Irken G, Akyol F, Cevik N T
Department of Pediatrics, Faculty of Medicine, Dokuz Eylül University, Izmir, Turkey.
Biol Neonate. 1995;68(2):100-3. doi: 10.1159/000244224.
In this study, using flow cytometry, we investigated the autofluorescence emitted by lymphocytes, monocytes and neutrophils exposed to different unconjugated bilirubin concentrations and investigated the relationship between these parameters. Different unconjugated bilirubin concentrations were prepared from a newborn serum with an unconjugated bilirubin concentration of 800 m mumol/l. The same concentrations of unconjugated bilirubin have been prepared from pure bilirubin. 10 microliters of cord blood were incubated at room temperature for 15 min with 90-microliter solutions with different bilirubin concentrations prepared from both serum and pure bilirubin. After incubation, cells were washed three times with PBS, erythrocytes were lysed by lysing buffer and run through flow cytometry immediately. Autofluorescence was measured by recording mean fluorescence channels for lymphocytes, monocytes and neutrophils. There was a good correlation between serum concentrations of unconjugated bilirubin that cells were exposed to and the autofluorescence intensity of neutrophils (r = 0.904, p < 0.005), monocytes (r = 0.759, p < 0.05) and lymphocytes (r = 0.766, p < 0.01). Results obtained with pure bilirubin were also similar. Autofluorescence emitted by lymphocytes was lower than that of monocytes (p < 0.01) or neutrophils (p < 0.0005).
在本研究中，我们使用流式细胞术，研究了暴露于不同浓度非结合胆红素的淋巴细胞、单核细胞和中性粒细胞发出的自发荧光，并研究了这些参数之间的关系。非结合胆红素的不同浓度是由未结合胆红素浓度为800 μmol/l的新生儿血清制备而成。相同浓度的非结合胆红素也由纯胆红素制备。将10微升脐血与由血清和纯胆红素制备的不同胆红素浓度的90微升溶液在室温下孵育15分钟。孵育后，细胞用磷酸盐缓冲液（PBS）洗涤三次，红细胞用裂解缓冲液裂解，然后立即进行流式细胞术检测。通过记录淋巴细胞、单核细胞和中性粒细胞的平均荧光通道来测量自发荧光。细胞所暴露的非结合胆红素血清浓度与中性粒细胞（r = 0.904，p < 0.005）、单核细胞（r = 0.759，p < 0.05）和淋巴细胞（r = 0.766，p < 0.01）的自发荧光强度之间存在良好的相关性。用纯胆红素获得的结果也相似。淋巴细胞发出的自发荧光低于单核细胞（p < 0.01）或中性粒细胞（p < 0.0005）。
