川楝素对H22肝癌荷瘤小鼠的抑瘤作用

Anticancer effects of the Chinese medicine toosendanin on the mice bearing H22 hepatocellular carcinoma

目的:建立荷瘤小鼠模型,探讨川楝素对肝癌的抑瘤作用及其机制。方法:建立H22肝癌移植瘤小鼠模型,随机分为0.9%氯化钠溶液对照组、环磷酰胺(cyclophosphamide,CTX)组(20mg/kg)、川楝素低剂量组(0.173mg/kg)和川楝素高剂量组(0.690mg/kg)共4组。各组药物处理后,测量小鼠体内肿瘤大小,观察肿瘤的生长曲线;剥瘤后称重,计算小鼠的肿瘤抑制率;行肿瘤组织病理形态学及HE染色观察,透射电子显微镜观察其超微结构改变;同时,观察川楝素对荷瘤小鼠心、肝、脾、肾、胸腺及睾丸组织的影响;免疫组织化学法检测肿瘤组织内Bcl-2、Bax和Fas蛋白的表达。结果:川楝素可显著抑制小鼠体内肿瘤的生长,川楝素低剂量组(0.173mg/kg)和高剂量组(0.690mg/kg)的抑瘤率分别为66.23%和87.01%(P<0.05);透射电子显微镜观察可见肿瘤组织超微结构中出现凋亡小体;HE染色显示小鼠心、肝、脾、肾及睾丸脏器形态正常,而胸腺组织中可见胸腺小叶的数量及面积减少甚至消失;免疫组织化学检测证实,小鼠肿瘤组织内Bcl-2蛋白表达下调,Bax和Fas蛋白表达上调(P<0.05)。结论:川楝素能够明显抑制小鼠移植瘤的生长,此作用可能与抑制肿瘤细胞增殖及促进肿瘤细胞凋亡有关。

Objective：To establish a mouse model bearing H22 hepatocellular carcinoma,and to explore the effects of the Chinese medicine toosendanin（TSN）on the solid carcinoma as well as its probable mechanisms.Methods：The BALB/c mice bearing H22 hepatocellular carcinoma were established and randomly divided into four groups：normal saline（NS）control group,low-dose（0.173 mg/kg）toosendanin（TSN）group,and high-dose（0.690 mg/kg）TSN group.The size of transplanted tumor was measured during treatment to draw the tumor growth volume curves in different groups.The transplanted tumors were excised after treatment and weighted to calculate the tumor growth inhibitory rates.The pathological histomorphology of transplanted tumor was observed under a light microscope after hematoxylin-eosin（HE）staining and then the ultramicro-structure changes were observed under a transmission electron microscope.The pathological features of the heart,liver,spleen,kidney,thymus and testis in tumor-bearing mice were observed.The expressions of Bcl-2,Bax and Fas were detected by immunohistochemistry.Results：The experiment in H22 tumor-bearing BALB/c mice documented TSN had obvious inhibitory effects on tumor growth.Compared with the control group,the tumor inhibition rates in low-dose TSN group and high-dose TSN group were 66.23% and 87.01%,respectively.Transmission electron microscopy showed that the tumor cells had apoptotic body-like morphology.The morphology of the liver,heart,spleen,kidney and testis tissues was normal and no significant difference in the morphology of these tissues between the control group and TSN group.Compared to the control group,the number and area of thymus lobules in TSN group and cyclophosphamide group were decreased or even absent.The expression of Bcl-2 protein was down-regulated,while the expressions of Bax and Fas proteins were up-regulated in tumor tissues treated with TSN compared with the control group.Conclusion：TSN can obviously inhibit the growth of transpanted tumor in the mice bearing hepatocellular carcinoma.The effect may be related to inhibiting proliferation and inducing apoptosis of tumor cells.