慢性乙肝患者PMNs内CXCL8及其受体CXCR1、CXCR2的表达

发布时间：2018-06-06 05:13

  本文选题：慢性乙肝 + 中性粒细胞　； 参考：《安徽理工大学》2014年硕士论文

【摘要】：目的：探讨慢性乙肝患者外周血CXCL8含量及中性粒细胞CXCL8、CXCR1、 CXCR2的表达水平。观察HBV侵染中性粒细胞后对CXCL8及其受体CXCR1、 CXCR2表达的影响。方法：筛选慢性乙肝患者42例,其中HBeAg阳性、阴性患者分别为17和25例,HBV-DNA阳性和阴性患者分别为13例和29例,以ELISA法检测患者血清CXCL8水平,Trizol提取PMNs总RNA,并逆转录为cDNA, qPCR法检测慢性乙肝PMNs内CXCL8、CXCR1、CXCR2mRNA表达量,以lgcDNA/lgGAPDH代表其最终mRNA水平。进一步观察血清CXCL8含量及PMNs内CXCL8、CXCR1、CXCR2mRNA表达水平与血清ALT水平的相互关系。采用SABC细胞免疫化学染色法观察患者外周血PMNs内CXCL8、CXCR1、CXCR2的表达,对比观察HBV侵染PMNs后对CXCL8、CXCR1、CXCR2表达的影响。 结果：慢性乙肝患者血清CXCL8含量为(432.43±216.33) pg/mL。其中, HBeAg(+)与HBeAg (-)患者CXCL8含量分别为(612.15±165.56) pg/mL和(310.21±152.42) pg/mL,差异有统计学意义(P0.01). HBV-DNA (+)与HBV-DNA (-)患者CXCL8含量分别为(593.74±174.03) pg/mL和(360.11±195.05) pg/mL,差异亦有统计学意义(P0.01)。患者外周血PMNs内CXCL8、CXCR1的mRNA水平分别为1.12±0.21、0.86±0.37,较正常对照组明显升高(P0.01)。其中HBeAg(+)患者CXCL8、CXCR1的mRNA水平分别为1.54±0.30、1.01±0.34,与HBeAg(-)组对照组相比,差异有显著性(P0.05);HBeAg(+)患者CXCR2的mRNA水平为1.21±0.15,与HBeAg(-)患者1.13±0.32相比,差异无统计学意义(P0.05)。HBV-DNA(+)组与HBV-DNA(-)组相比,CXCL8、CXCR1的mRNA水平分别为1.22±0.26、1.05±0.21与1.08±0.18、0.79±0.42,差异有显著性(P0.05)；CXCR2的mRNA水平分别为1.13±0.25和1.17±0.29,差异无统计学意义(P0.05)。血清CXCL8水平与ALT异常升高呈正相关0=0.859, P0.01), PMNs内CXCL8. CXCR1的mRNA与ALT异常升高亦有一定相关性(r=0.688,P0.01；r=0.585,P0.01),而CXCR2表达与ALT异常升高无明显相关性(r=0.088,P0.05)。SABC免疫细胞化学染色结果显示,CXCL8主要位于PMNs胞浆中,CXCR1、 CXCR2多见于胞浆和胞膜上。其中HBeAg(+)者CXCL8、CXCR1免疫着色较深,而CXCR2免疫着色较浅；PMNs内HBVDNA(+)者CXCL8、CXCR1免疫着色亦较深,而CXCR2免疫着色亦较浅。与对照组相比,CXCL8、CXCR1的表达水平差异均有显著性(P0.05),而CXCR2的表达无统计学意义(P0.05)。 结论：HBV除对肝细胞具有较高亲嗜性,还能侵染中性粒细胞,形成肝外潜伏或隐匿感染,此可能是导致慢性乙肝病情迁延反复、久治难愈的重要病因。慢性乙肝患者外周血CXCL8含量增高,PMNs内CXCL8、CXCR1、CXCR2的mRNA水平增高,其中,血清CXCL8水平及CXCL8、CXCR1的mRNA与血清ALT含量呈正相关。HBV侵染中性粒细胞后可干扰宿主细胞正常代谢,使CXCL8分泌增加,细胞膜上CXCR1、CXCR2表达增强。CXCL8主要位于PMNs胞浆中,CXCR1、 CXCR2多见于胞浆和胞膜上,其着色程度与患者HBeAg表达、HBV DNA载量密切相关。HBV侵染中性粒细胞后可使胞内分泌释放CXCL8进一步增加,胞膜CXCR1表达进一步增强。高表达CXCR1的中性粒细胞又与CXCL8相互作用,趋化吸引更多中性粒细胞至病灶,参与局部炎性损伤和组织修复。
[Abstract]:Objective: To investigate the CXCL8 content of peripheral blood and the expression level of CXCL8, CXCR1 and CXCR2 of neutrophils in patients with chronic hepatitis B. The effect of HBV on the expression of CXCL8 and its receptor CXCR1 and CXCR2 after HBV infection was observed. Methods: 42 cases of chronic hepatitis B were screened, of which 17 and 25 negative patients were positive and negative, and HBV-DNA positive and negative. The patients were 13 cases and 29 cases respectively. The serum CXCL8 level was detected by ELISA. The total RNA of PMNs was extracted by Trizol and cDNA. The qPCR method was used to detect the CXCL8, CXCR1, CXCR2mRNA expression in PMNs. The relationship between serum ALT level and the expression of CXCL8, CXCR1 and CXCR2 in peripheral blood PMNs were observed by SABC cell immunochemistry staining, and the effects of HBV infection on CXCL8, CXCR1, and CXCR2 expression were observed.
Results: the serum CXCL8 content of patients with chronic hepatitis B was (432.43 + 216.33) pg/mL., and the CXCL8 content of HBeAg (+) and HBeAg (-) patients was (612.15 + 165.56) pg/mL and (310.21 + 152.42) pg/mL respectively, the difference was statistically significant (P0.01). The CXCL8 content of HBV-DNA (+) and HBV-DNA (-) patients was (593.74 + 174.03) pg/mL and (360.11 + 195.05) respectively. G/mL, the difference was also statistically significant (P0.01). The level of CXCL8 and CXCR1 in the peripheral blood of the patients was 1.12 + 0.21,0.86 + 0.37, respectively, which was significantly higher than that in the normal control group (P0.01). The HBeAg (+) patients' CXCL8, CXCR1 mRNA level was 1.54 + 0.34, respectively, and the difference was significant compared with the control group (+). The mRNA level of CXCR2 was 1.21 + 0.15, compared with 1.13 + 0.32 of HBeAg (-) patients, the difference was not statistically significant (P0.05) in.HBV-DNA (+) group, the mRNA level of CXCL8, CXCR1 was 1.22 + 0.26,1.05 + 0.21 and 1.08 + 0.18,0.79 + 0.42 respectively, and the difference was significant (P0.05), and the levels were 1.13 + 0.25 and 1.17 + 0.29, respectively. There was no statistical significance (P0.05). The level of serum CXCL8 was positively correlated with the abnormal increase of ALT, 0=0.859, P0.01), and there was a correlation between mRNA and ALT abnormal increase of CXCL8. CXCR1 in PMNs. CL8 is mainly located in the cytoplasm of PMNs, and CXCR1 and CXCR2 are more common in the cytoplasm and cell membrane. Among them, HBeAg (+) is CXCL8, CXCR1 immunologic coloring is deeper, CXCR2 immune coloring is shallow, PMNs HBVDNA (+) CXCL8, CXCR1 immune coloring is also deep, and the immune coloring is also shallow. However, the expression of CXCR2 was not statistically significant (P0.05).
Conclusion: HBV has high affinity to liver cells, but also can infect neutrophils and form extrahepatic latent or concealed infection. This may be an important cause of chronic hepatitis B relapse and prolonged treatment. The content of CXCL8 in peripheral blood of chronic hepatitis B patients is higher, and the level of mRNA in CXCL8, CXCR1 and CXCR2 in PMNs is higher, in which the serum CXCL8 water is increased. CXCL8, CXCR1 mRNA and serum ALT content positively correlated with.HBV infection of neutrophils, which can interfere with normal metabolism of host cells and increase the secretion of CXCL8. The expression of CXCR1 and CXCR2 on the cell membrane is mainly in the PMNs cytoplasm, CXCR1, CXCR2 is mostly seen in the cytoplasm and the cytoplasm, and the degree of coloring is closely related to the patient's expression. After related.HBV infection of neutrophils, the endocrine release of CXCL8 can be further increased and the expression of CXCR1 in the cell membrane is further enhanced. The neutrophils with high expression of CXCR1 interact with CXCL8, and chemotaxis attract more neutrophils to the focus, and participate in local inflammatory injury and tissue repair.
【学位授予单位】：安徽理工大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R512.62

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