重组人粒细胞集落刺激因子对急性肝衰竭大鼠的作用

发布时间：2018-06-18 13:34

本文选题：重组人粒细胞集落刺激因子D-氨基半乳糖 + 急性肝衰竭　；参考：《天津医科大学》2013年硕士论文

【摘要】：目的评价重组人粒细胞集落刺激因子(rhG-CSF)对于D-氨基半乳糖(D-GalN)诱导的急性肝衰竭大鼠的影响。方法把雄性SD大鼠随机分为正常对照组、肝衰模型组、rhG-CSF立即给药组、rhG-CSF延迟给药组。除正常对照组外,其余三组均腹腔内注射D-GalN (1400mg/kg)建立大鼠急性肝衰竭模型。RhG-CSF立即给药组于造模后2小时立即给予皮下注射rhG-CSF (50μg/kg), rhG-CSF延迟给药组于造模后72小时给予皮下注射rhG-CSF (50μg/kg),以上两组分别于造模后72小时、2小时各接受一次皮下注射生理盐水。肝衰模型组于造模后2小时、72小时给予两次皮下注射生理盐水。为观察rhG-CSF对急性肝衰竭大鼠不同时间点的影响,于造模后12、24、48、72、120小时,分别处死肝衰模型组及rhG-CSF立即给药组5只大鼠,取血检测丙氨酸氨基转移酶、总胆红素、外周血白细胞,取肝组织做HE染色观察肝脏病理变化,并应用免疫组织化学方法测定PCNA阳性细胞、TNF-α阳性细胞；以上两组均取10只大鼠观察造模后120小时生存率。RhG-CSF延迟给药组以皮下注射rhG-CSF为观察始点,以肝衰造模120小时为观察终点,并与肝衰模型组、rhG-CSF立即给药组进行比较。结果肝脏病理结果显示：RhG-CSF立即给药组与肝衰模型组相比,除肝衰造模120小时外,其余各个时间点病理结果均显示肝细胞坏死更为严重,同时伴有更为明显的胆管再生；肝衰造模后120小时的病理结果显示,rhG-CSF立即给药组较肝衰模型组肝脏组织结构恢复较好伴有更少的淋巴细胞浸润,而rhG-CSF延迟给药组其肝脏组织结构恢复又明显好于rhG-CSF立即给药组。免疫组织化学方法结果显示：RhG-CSF立即给药组PCNA阳性细胞数、TNF-a阳性细胞数于各个时间点,水平均明显高于肝衰模型组;而rhG-CSF延迟给药组于造模后120小时,PCNA阳性细胞数明显高于肝衰模型组,但相比于rhG-CSF立即给药组无明显差别,TNF-a阳性细胞数相比于肝衰模型组、rhG-CSF立即给药组则均明显减少。血清丙氨酸氨基转移酶及总胆红素水平结果显示：RhG-CSF立即给药组在肝衰造模后各个时间点其水平均高于肝衰模型组,但血清丙氨酸氨基转移酶仅于造模后24小时两组差异有统计学意义(P0.05),总胆红素水平仅于造模后12,24小时差异有统计学意义(P0.05);RhG-CSF延迟给药组于造模后120小时,血清丙氨酸氨基转移酶及总胆红素水平与肝衰模型组及rhG-CSF立即给药组相比差异均无统计学意义。外周血白细胞计数结果显示：于造模后各个时间点,rhG-CSF立即给药组均高于肝衰模型组,除造模后12小时外,其余时间点差异均有统计学意义(P0.05)；RhG-CSF延迟给药组于造模后120小时,外周血白细胞计数明显高于肝衰模型组,差异有统计学意义(P0.05),但与rhG-CSF立即给药组相比,外周血白细胞计数较之减少,差异无统计学意义(P0.05)。RhG-CSF立即给药组和肝衰模型组相比,生存率差异无统计学意义(30%,78.9+8.95小时VS20%,77.30±10.18小时)；RhG-CSF延迟给药组6只成功给予rhG-CSF大鼠中5只存活。结论在ALF急性期应用rhG-CSF加重肝脏炎症反应；在急性期过后应用rhG-CSF促进肝脏组织的修复,降低炎症反应,并有提高ALF生存率的可能,结果有待进一步研究证实。
[Abstract]:Objective to evaluate the effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) on D- galactoside (D-GalN) - induced acute liver failure in rats.
Methods the male SD rats were randomly divided into the normal control group, the liver failure model group, the rhG-CSF immediate administration group and the rhG-CSF delayed administration group. Except the normal control group, the other three groups were intraperitoneally injected with D-GalN (1400mg/kg) to establish the rat acute hepatic failure model.RhG-CSF immediately given the subcutaneous injection of rhG-CSF (50 mu g/kg) immediately after the model. The rhG-CSF delayed injection group was given a subcutaneous injection of rhG-CSF (50 g/kg) 72 hours after the model, and the above two groups were given a subcutaneous injection of saline at 72 hours after making the model, and the liver failure model group was given two subcutaneous injection of saline after 72 hours after the model of the model, in order to observe the difference between the rhG-CSF and the rats with acute hepatic failure. At 12,24,48,72120 hours after the model, 5 rats were sacrificed to the liver failure model group and the rhG-CSF immediate administration group. The serum alanine aminotransferase, total bilirubin, peripheral blood leukocytes and liver tissue were detected by HE staining, and the pathological changes of the liver were observed by HE staining. The PCNA positive cells were detected by the immunoimmunoassay method, and the positive of TNF- alpha was positive. Cells in the above two groups were taken 10 rats to observe the 120 hour survival rate of.RhG-CSF delayed injection group by subcutaneous injection of rhG-CSF as the beginning point, with liver failure model for 120 hours as the observation end point, and compared with the liver failure model group, rhG-CSF immediate drug delivery group.
Results the liver pathological results showed that the RhG-CSF group was compared with the liver failure model group. Except for the liver failure model 120 hours, the pathological results of the other time points showed that the hepatocyte necrosis was more serious and accompanied by more obvious bile duct regeneration. The pathological results of the 120 hours after the liver failure model showed that the rhG-CSF immediate administration group was more than the liver failure. The liver tissue structure of the model group was well restored with less lymphocyte infiltration, and the liver tissue structure of the rhG-CSF delayed drug group was better than that of the rhG-CSF immediate administration group. The results of immunohistochemical staining showed that the number of PCNA positive cells in the group of RhG-CSF was immediately given, and the number of TNF-a positive cells was at every time point, and the level was obvious. The number of PCNA positive cells in the rhG-CSF delayed administration group was significantly higher than that in the liver failure model group at 120 hours after the model group, but the number of TNF-a positive cells was significantly lower than that in the rhG-CSF group. The serum alanine aminotransferase and total bilirubin were significantly reduced in the rhG-CSF group. The level results showed that the level of RhG-CSF immediately after the liver failure model was higher than that in the liver failure model group, but the serum alanine aminotransferase was statistically significant (P0.05) only 24 hours after the model (P0.05), and the total bilirubin level was statistically significant (P0.05) only after the model of the model (P0.05); RhG-CSF delayed administration. There was no significant difference in serum alanine aminotransferase and total bilirubin level between the liver failure model group and the rhG-CSF group at 120 hours after the model group. The results of the peripheral blood white blood cell count showed that the rhG-CSF immediate administration group was higher than the liver failure model group at each time point after the model building, except 12 hours after the model building. The time point difference was statistically significant (P0.05); the white blood cell count in peripheral blood was significantly higher than that in the liver failure model group at 120 hours after the RhG-CSF delayed delivery group, and the difference was statistically significant (P0.05), but compared with the immediate rhG-CSF group, the white blood cell count of the peripheral blood was less than that of the rhG-CSF group, and the difference was not statistically significant (P0.05).RhG-CSF immediately given the drug There was no significant difference in the survival rate between the group and the liver failure model group (30%, 78.9+8.95 hours VS20%, 77.30 + 10.18 hours), and 6 of the RhG-CSF delayed drug delivery groups were given 5 survival in the rhG-CSF rats.
Conclusion the use of rhG-CSF in the acute phase of ALF aggravates the liver inflammation, and the application of rhG-CSF to promote the repair of liver tissue, reduce the inflammatory response and improve the survival rate of ALF after the acute stage. The results need to be further confirmed.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R575.3

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