泡球蚴囊液对大鼠肝星状细胞MAPK信号通路影响的初步研究

发布时间：2018-07-16 08:02

【摘要】：目的探讨泡球蚴囊液对大鼠肝星状细胞丝裂素活化蛋白激酶(mitogenactivated protein kinas，MAPK)信号通路的影响。方法采用大鼠肝星状细胞HSC-T6与不同蛋白浓度的泡球蚴囊液共培养，共11组，浓度分别为13.5mg/mL的泡球蚴囊液原液，6.8mg/mL、3.4mg/mL、1.7mg/mL、0.9mg/mL、0.4mg/mL、0.2mg/mL、0.1mg/mL、0.05mg/mL、0.025mg/mL、0.01mg/mL。培养24h后收集细胞,同时收集对照组(无泡球蚴囊液干预)细胞。利用荧光实时定量PCR检测ERK1/2、JNK1/2和p38MAPK的变化。结果泡球蚴原囊液与HSC共培养24h后大部分细胞发生固缩，呈脱落前兆；1/2原囊液组部分细胞固缩，呈脱落前兆，而部分HSC-T6细胞收缩成长梭形，细胞生出许多细长的伪足，是正常状态的2倍以上；1/4原囊液组部分细胞呈收缩的长梭形，生出许多细长的伪足，但大部分细胞仍呈现为贴壁的胞体较大的不规则多边形，且伸出较短的伪足与其他细胞连接成片状；1/8原囊液以下组与空白对照组细胞从形态上则无差异。实时荧光定量检测显示ERK1/2、 JNK1/2和p38MAPK mRNA水平在泡球蚴囊液0.4mg/mL时显著增加，在6.8mg/mL时相对于对照组高表达，与对照组比较有显著性差异(P0.05)。结论不同浓度泡球蚴囊液均可通过与大鼠肝星状细胞表面受体结合激活大鼠肝星状细胞MAPK信号通路，，其中浓度为6.8mg/mL的泡球蚴囊液干预结果较为显著。
[Abstract]:Objective to investigate the effect of hydatid cyst fluid on mitogen-activated protein kinase (mitogenactivated protein) signaling pathway in rat hepatic stellate cells. Methods Rat hepatic stellate cells HSC-T6 were co-cultured with hydatid cyst fluid of different protein concentrations. In 11 groups, 6.8mg / mL1.7mg / mL 0.2mg / mL 0.1 mg / mL ~ 0.025mg / mL ~ 0.025 mg / mL ~ (-1) 0.025mg / mL ~ (-1) mg / mL ~ (-1), respectively, in 11 groups of rat hepatic stellate cells (HSC-T6) were co-cultured with different protein concentrations. The concentration of HSC-T6 was 6.8mg / mL ~ (-1) mg / mL ~ (-1) ~ (-1) mg / mL ~ (-1) 0.2mg / mL ~ (0.1) mg / mL ~ (-1). Cells were collected after 24 hours of culture and control group (without the intervention of alveolar hydatid fluid). The changes of ERK1 / 2 JNK1 / 2 and p38 MAPK were detected by fluorescence real time quantitative PCR. Results after 24 hours of co-culture with HSC, most of the cells of Echinococcus alveolaris had pyknosis, and some of the cells in the group of 1 / 2 / 2 of the procytial fluid showed pyknosis, while some of the HSC-T6 cells contracted and formed fusiform, and the cells produced many slender pseudopodia. More than 2 times the normal state, some of the cells in the primordial fluid group showed long spindle shape of contraction, producing many slender pseudopods, but most of the cells still showed large irregular polygons attached to the wall of the cell body. And there was no difference in morphology between the group with short pseudopodia and other cells connecting to the other cells to form a flake of 1 / 8 procapsular fluid and the blank control group. Real-time fluorescence quantitative analysis showed that ERK1 / 2, JNK1 / 2 and p38 MAPK mRNA levels were significantly increased at 0.4 mg / mL in hydatid cyst fluid, and were significantly higher than those in control group at 6.8 mg / mL compared with control group (P0.05). Conclusion different concentrations of hydatid cyst fluid can activate MAPK signaling pathway of rat hepatic stellate cells by binding to rat hepatic stellate cell surface receptor.
【学位授予单位】：青海大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R532.32

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