高良姜素通过巨噬细胞极化及抑制锌指蛋白Zbed3诱导大鼠肝癌细胞凋亡的研究

Effect of galangin on apoptosis of rat hepatocellular carcinoma cells induced by macrophage polarization and inhibition of zinc finger protein Zbed3

目的 探讨高良姜素诱导巨噬细胞极化对肝癌大鼠瘤体体积及锌指蛋白Zbed3的影响。方法 健康6~8周龄SD雄性大鼠74只,体质量250~300 g,常规饲养,自由饮食饮水。随机分为对照组(健康大鼠常规饲养,n=14)、模型组(模型大鼠常规饲养,n=13)、高良姜素低剂量组(模型+25 mg/kg/d高良姜素,n=13)、高良姜素中剂量组(模型+50 mg/kg/d高良姜素,n=13)及高良姜素高剂量组(模型+100 mg/kg/d高良姜素,n=13)。HE染色检测肝组织病理形态,TUNEL检测肝癌细胞凋亡,游标卡尺测定肿瘤体积,免疫组化检测CD11c、CD206表达,Wbstern Blot检测Zbed3、GSK3β、Axin、β-catenin、c-myc、cyclin-D1水平。结果 对照组大鼠肝小叶结构清晰,肝细胞排列有序,形态正常;模型组大鼠瘤组织呈膨胀性生长,癌细胞大小不一,肝细胞索挤压,排列不齐,并可见纤维素样结构;高良姜素低、中、高剂量组大鼠均出现肿瘤细胞间隙增宽,肿瘤细胞出现不同程度的肿胀变性,呈现泡沫状改变,肿瘤细胞巢可见大量炎细胞浸润,各组均可见不同程度的肿瘤细胞坏死灶。与对照组相比,模型组大鼠CD11c、GSK3β、Axin降低,CD206、Zbed3、β-catenin、c-ymc、cyclin-D1升高(P<0.05)。与模型组相比,高良姜素低剂量组肝癌细胞凋亡率、CD11c、GSK3β、Axin升高,瘤体体积、CD206、Zbed3、β-catenin、c-ymc、cyclin-D1降低(P<0.05)。与高良姜素低剂量组相比,高良姜素中剂量组大鼠肝癌细胞凋亡率、CD11c、GSK3β、Axin升高,瘤体体积、CD206、Zbed3、β-catenin、c-ymc、cyclin-D1降低(P<0.05)。与高良姜素中剂量组相比,高良姜素高剂量组大鼠肝癌细胞凋亡率、CD11c、GSK3β、Axin升高,瘤体体积、CD206、Zbed3、β-catenin、c-ymc、cyclin-D1降低(P<0.05)。结论 高良姜素通过调控Wnt/β-catenin通路,促进巨噬细胞向M1型转化并抑制其向M2型转化,降低了Zbed3水平从而促进癌细胞凋亡并减小瘤体体积。

Objective This paper investigated the effect of galangin-induced macrophage polarization on tumor volume and zinc finger protein Zbed3 in rats with liver cancer.Methods Seventy-four healthy male SD rats aged 6 to 8 weeks with body weight of 250 to 300 g were fed conventionally with free diet and water.The parameters were randomly divided into control group(normal feeding of healthy rats,n=14),model group(model rats were reared routinely,n=13),low dose of galangin(model+25 mg/kg/d galangin,n=13),medium dose group of golangin(model+50 mg/kg/d golangin,n=13),high-dose group of golangin(model+100 mg/kg/d golangin,n=13).HE staining was used to detect the pathological morphology of liver tissue.Apoptosis of hepatoma cells was detected by TUNEL.Tumor volume was measured with vernier calipers.The expression of CD11c and CD206 was detected by immunohistochemistry.The levels of Zbed3,GSK3β,Axin,β-catenin,C-myc and cyclin-D1 were detected by Western Blot.Results Rats in control group had clear hepatic lobule structure and normal liver cells.The tumor tissue of the model group showed dilatant growth,cancer cells were of different sizes,and the cords of liver cells were extruded and disarranged,with fibrin-like structure.In low,medium and high dose galangin groups,tumor cell space was widened,tumor cells showed swelling and foamy changes to varying degrees,tumor cell nests were infiltrated by a large number of inflammatory cells,and tumor cell necrosis foci of varying degrees were observed in all groups.Compared with control group,CD11c,GSK3βand Axin in model group were decreased,and CD206,Zbed3,β-catenin,C-YMC and cyclin-D1 were increased(P<0.05).Compared with model group,apoptosis rate,CD11c,GSK3βand Axin were increased,and tumor volume,CD206,Zbed3,β-catenin,C-YMC and cyclin-D1 were decreased in GF group(P<0.05).Compared with low dose group,apoptosis rate,CD11c,GSK3βand Axin were increased,and tumor volume,CD206,Zbed3,β-catenin,C-YMC and Pressume cyclin-D1 were decreased in medium dose group(P<0.05).Compared with medium dose group,apoptosis rate,CD11c,GSK3βand Axin were increased in high dose group,and tumor volume,CD206,Zbed3,β-catenin,C-YMC and cyclin-D1 were decreased(P<0.05).Conclusion By regulating the Wnt/β-catenin pathway,galangin promoted the transformation of macrophages into M1-type and inhibited the transformation of macrophages into M2-type,and reduced the level of Zbed3,thus promoting the apoptosis of cancer cells and reducing the tumor volume.