论文部分内容阅读
目的研究三七总皂甙(PNS)对大鼠肝卵圆细胞(HOC)增殖模型肝脏p-ERK1/2表达的影响,探讨PNS调节HOC增殖的可能信号途径。方法将66只健康雄性Wistar大鼠随机分成对照组(n=6)、模型组(n=30)和PNS组(n=30)。模型组、PNS组采用2-AFF/PH方法建立HOC增殖模型,PNS组大鼠建模时(即第一次2-AAF灌胃前1 h)予以PNS 25 mg/(kg.d)腹腔内注射,并持续至实验结束。模型组、PNS组在术后第4小时,第4、8、12、16天随机取6只大鼠检测。对照组大鼠未予特殊处理。采用组织病理技术观察肝组织的形态学变化;免疫组织化学和细胞形态学方法计数HOC;免疫组织化学及Western blot分析p-ERK1/2蛋白水平。结果对照组、模型组和PNS组术后第4小时未见HOC增殖。模型组术后第4天汇管区有HOC增殖反应,第8天HOC增殖达峰值,第12天HOC从汇管区向肝实质内浸润,第16天HOC增殖较第12天减少。与模型组比较,PNS组第4、8、16天HOC增殖均减少(均P<0.05),第12天HOC增殖增多(P<0.05)。对照组大鼠肝组织p-ERK1/2蛋白水平很低,模型组p-ERK1/2蛋白表达术后各时点分别为对照组的(2.24±0.17)、(3.87±0.35)(、5.28±0.48)、(2.96±0.33)、(2.12±0.19)倍;PNS组各时点p-ERK1/2的表达分别为对照组的(1.56±0.23)、(2.77±0.19)、(3.53±0.35)、(3.62±0.45)(、1.36±0.16)倍。与模型组比较,PNS组术后第12天p-ERK1/2表达升高(P<0.01),第4小时,第4、8、16天表达均减少(均P<0.05)。结论在HOC介导肝再生过程中,HOC的p-ERK1/2表达与其增殖趋势相同;PNS可使大鼠肝脏的HOC增殖及p-ERK1/2的表达降低、滞后、持续时间延长。
Objective To investigate the effect of panax notoginseng saponins (PNS) on the expression of hepatic p-ERK1/2 in rat hepatic oval cell (HOC) proliferation model, and to explore the possible signal pathways of PNS regulating HOC proliferation. Methods Sixty-six healthy male Wistar rats were randomly divided into control group (n=6), model group (n=30) and PNS group (n=30). The HOC proliferation model was established using 2-AFF/PH method in the model group and the PNS group. PNS 25 mg/(kg.d) was intraperitoneally injected into the PNS group when the rats were modeled (ie, 1 h before the first 2-AAF administration). Inject and continue until the end of the experiment. In the model group and the PNS group, 6 rats were randomly selected on the 4th, 4th, 8th, 12th and 16th days after operation. Rats in the control group were not treated specifically. Histopathological techniques were used to observe the morphological changes of liver tissues; HOC was counted by immunohistochemistry and cytomorphology methods; p-ERK1/2 protein levels were analyzed by immunohistochemistry and Western blot. Results There was no HOC proliferation at 4 hours after operation in the control, model, and PNS groups. On the 4th day after operation, the HOC proliferative response was found in the portal area of the model group. On the 8th day, HOC proliferation peaked. On the 12th day, HOC infiltrated into the parenchymal hepatic parenchyma from the portal area. On the 16th day, HOC proliferation decreased compared with the 12th day. Compared with the model group, the proliferation of HOC on the 4th, 8th and 16th days of the PNS group decreased (all P<0.05), and on the 12th day, HOC proliferation increased (P<0.05). The level of p-ERK1/2 protein in the liver of control rats was very low. The expression of p-ERK1/2 protein in the model group was (2.24±0.17), (3.87±0.35) (5.28±), respectively. 0.48), (2.96±0.33), (2.12±0.19) times; the expression of p-ERK1/2 at each time point in the PNS group was (1.56±0.23), (2.77±0.19), (3.53±0.35) in the control group; , (3.62 ± 0.45) (, 1.36 ± 0.16) times. Compared with the model group, the expression of p-ERK1/2 was increased on the 12th day after operation in the PNS group (P<0.01), and decreased on the 4th, 4th, 8th, and 16th days (P<0.05). Conclusions HOC p-ERK1/2 expression is similar to that of HOC in the process of liver regeneration mediated by HOC. PNS induces the decrease of HOC proliferation and p-ERK1/2 expression, and the prolongation of lag time and duration of rat liver.