【摘要】：目的探討泡球蚴囊液對(duì)大鼠肝星狀細(xì)胞絲裂素活化蛋白激酶(mitogenactivated protein kinas，MAPK)信號(hào)通路的影響。 方法采用大鼠肝星狀細(xì)胞HSC-T6與不同蛋白濃度的泡球蚴囊液共培養(yǎng)，共11組，濃度分別為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。培養(yǎng)24h后收集細(xì)胞,同時(shí)收集對(duì)照組(無(wú)泡球蚴囊液干預(yù))細(xì)胞。利用熒光實(shí)時(shí)定量PCR檢測(cè)ERK1/2、JNK1/2和p38MAPK的變化。 結(jié)果泡球蚴原囊液與HSC共培養(yǎng)24h后大部分細(xì)胞發(fā)生固縮，呈脫落前兆；1/2原囊液組部分細(xì)胞固縮，呈脫落前兆，而部分HSC-T6細(xì)胞收縮成長(zhǎng)梭形，細(xì)胞生出許多細(xì)長(zhǎng)的偽足，是正常狀態(tài)的2倍以上；1/4原囊液組部分細(xì)胞呈收縮的長(zhǎng)梭形，生出許多細(xì)長(zhǎng)的偽足，但大部分細(xì)胞仍呈現(xiàn)為貼壁的胞體較大的不規(guī)則多邊形，且伸出較短的偽足與其他細(xì)胞連接成片狀；1/8原囊液以下組與空白對(duì)照組細(xì)胞從形態(tài)上則無(wú)差異。 實(shí)時(shí)熒光定量檢測(cè)顯示ERK1/2、 JNK1/2和p38MAPK mRNA水平在泡球蚴囊液0.4mg/mL時(shí)顯著增加，在6.8mg/mL時(shí)相對(duì)于對(duì)照組高表達(dá)，與對(duì)照組比較有顯著性差異(P0.05)。 結(jié)論不同濃度泡球蚴囊液均可通過(guò)與大鼠肝星狀細(xì)胞表面受體結(jié)合激活大鼠肝星狀細(xì)胞MAPK信號(hào)通路，，其中濃度為6.8mg/mL的泡球蚴囊液干預(yù)結(jié)果較為顯著。
[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.
【學(xué)位授予單位】：青海大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R532.32

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