没药甾酮对二乙基亚硝胺诱导大鼠肝癌的抑制作用及其机制

摘要/Abstract

摘要： 目的探究没药甾酮(GS)对二乙基亚硝胺(DEN)诱导大鼠肝细胞癌(HCC)的抑制作用及其机制。方法构建DEN诱导的SD大鼠肝细胞癌模型,将造模成功大鼠随机分为HCC组和GS组,选择自由喂养的大鼠为对照组,每组6只。GS组以50 mg/kg GS腹腔注射,连续注射4周,对照组和HCC组注射等量0.9% NaCl溶液。观察大鼠的肝脏大体形态并测量体质量、肝质量和肝体质量比; HE染色法检测大鼠肝脏病理变化;免疫组织化学和蛋白质印迹检测p53相关线粒体凋亡信号通路蛋白p53、Bax、Bcl-2、caspase-3、caspase-9的表达水平。结果与对照组大鼠体质量(501.04 ± 18.03)g相比,HCC组(406.07 ± 22.91)g和GS组(357.83 ± 29.01)g均显著下降(P<0.001)。HCC组肝质量和肝体质量分别为(27.68 ± 6.34)g、(6.88 ± 1.87)g,GS组分别为(18.25 ± 2.20)g、(5.13 ± 0.72)g,均显著高于对照组[分别为(13.67 ± 1.40)g、(2.74 ± 0.38)g],且GS组低于HCC组(P=0.002、0.003)。与对照组相比,肉眼可见HCC组肝脏体积变大、表面多个癌结节呈弥漫分布,GS组亦可见多个呈弥漫分布的癌结节,但较HCC组,癌结节明显减少;镜下可见HCC组肝细胞异型增生、局灶结节性增生现象,而GS组的肝细胞异型增生坏死及局灶结节性增生则明显改善。免疫组化结果显示,GS组p53(56.42 ± 6.84)、Bax(43.02 ± 5.27)、caspase-3(13.37 ± 2.50)蛋白表达水平均显著高于对照组(19.33 ± 2.98、16.07 ± 3.22、3.42 ± 1.04)和HCC组(39.32 ± 5.99、27.42 ± 3.74、9.63 ± 1.53),且3组比较差异均有统计学意义(P<0.05);而Bcl-2(8.52 ± 3.31)蛋白表达水平显著低于对照组(22.03 ± 3.74)和HCC组(15.38 ± 2.10),3组比较差异均有统计学意义(P<0.05);3组caspase-9(1.27 ± 0.71、1.07 ± 0.70、1.43 ± 0.81)蛋白表达水平差异均无统计学意义(P>0.05)。蛋白质印迹亦得出来相似的结果。结论 GS对DEN诱导的大鼠HCC有一定的抑制作用,其作用机制可能与p53相关线粒体凋亡信号通路有关。

关键词: 没药甾酮, 肝细胞癌, p53, 二乙基亚硝胺

Abstract: Objective To explore the inhibitory effect and its mechanism.of guggulsterone (GS) on diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC) in rats. Methods DEN-induced HCC model was constructed in SD rats. Successful HCC modeling rats were randomly divided into the HCC group (n=6) and the GS group (n=6, 50 mg/kg), and free-feeding rats were randomly selected as the control group (n=6). The GS group was continuously injected with GS for four weeks, while the control group and HCC group were injected with an equal amount of physiological saline. The liver gross morphology was observed and the body mass, liver mass and liver-body mass ratio were measured in rats. Pathological changes of rat liver were detected by hematoxylin and eosin (HE) Staining staining, and the expression levels of p53-associated mitochondrial apoptotic signaling pathway proteins, p53, B-cell lymphoma 2 (Bcl-2), BCL2-associated X protein (Bax), caspase-3, and caspase-9 were detected by immunohistochemistry staining and Western blot methods, respectively. Results Compared with the control group (501.04 ± 18.03) g, the body mass of rats was significantly decreased in both HCC (406.07 ± 22.91) g and GS groups [(357.83 ± 29.01)g, P<0.001]. The liver mass and liver body mass ratio of the HCC group (27.68 ± 6.34, 6.88 ± 1.87) g and the GS group (18.25 ± 2.20, 5.13 ± 0.72) g were significantly higher than those of control group (13.67 ± 1.40, 2.74 ± 0.38) g, and the GS group was lower than the HCC group (P=0.002, P=0.003). The liver of the HCC group was enlarged with multiple cancerous nodules diffusely distributed on the surface, and multiple cancerous nodules diffusely distributed were also seen in the GS group, but the number of cancerous nodules was significantly reduced compared with that in the HCC group. Microscopically, the phenomena of hepatocellular heterogeneous hyperplasia and focal nodular hyperplasia were seen in the HCC group, whereas hepatocellular heterogeneous hyperplasia necrosis and focal nodular hyperplasia were significantly improved in the GS group. Immunohistochemistry showed that the protein expression levels of p53 (56.42 ± 6.84), Bax (43.02 ± 5.27) and caspase-3 (13.37 ± 2.50) in the GS group were significantly higher than those in the control group (19.33 ± 2.98, 16.07 ± 3.22, 3.42 ± 1.04) and the HCC group (39.32 ± 5.99, 27.42 ± 3.74, 9.63 ± 1.53), and there was a statistically significant difference in the comparison of the three groups (P<0.05), while the protein expression level of Bcl-2 (8.52 ± 3.31) was significantly lower than that in the control group (22.03 ± 3.74) and the HCC group (15.38 ± 2.10), and there was a statistically significant difference among the comparison of the three groups (P<0.05). The caspase-9 protein expression level was not statistically different among the three groups (1.27 ± 0.71, 1.07 ± 0.70, 1.43 ± 0.81, P>0.05). Similar results were obtained by Western blot. Conclusion GS inhibited DEN-induced HCC, which may be related to the p53-related mitochondrial apoptotic signaling pathway.

Key words: Guggulsterone, Hepatocellular carcinoma, P53, Diethylnitrosamine

刘雄涛, 王怡恺, 薛鹏军, 康沛, 张欣, 石娟娟. 没药甾酮对二乙基亚硝胺诱导大鼠肝癌的抑制作用及其机制[J]. 肝脏, 2025, 30(9): 1225-1229.

LIU Xiong-tao, WANG Yi-kai, XUE Peng-jun, KANG Pei, ZHANG Xin, SHI Juan-juan. Inhibitory effect and its mechanism of guggulsterone on diethylnitrosamine-induced hepatocellular carcinoma in rats[J]. Chinese Hepatolgy, 2025, 30(9): 1225-1229.

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