栽培杨黄不同提取物对H22荷瘤小鼠的抗肿瘤作用及机理

Antitumor effect and mechanism of different extracts of cultivated Phellinus vaninii on H22 tumor bearing mice

摘要:

为探讨栽培杨黄子实体不同提取物对H₂₂荷瘤小鼠的抑瘤作用及机制，将150只ICR小鼠随机分为空白组、模型组、CTX组、杨黄水提取物组、乙醇提取物组、石油醚提取物组及粗多糖组。采用H₂₂肝癌细胞建立实体瘤模型，给药第10天处死小鼠，计算脾、胸腺脏器指数及抑瘤率，检测血清中TNF-α、INF-γ、VEGF及苏木精-伊红和免疫组化染色法观察肿瘤组织病理变化，同时Western blotting检测肿瘤相关蛋白表达。结果表明，石油醚提取物的高剂量组抑瘤率(73.21%)最好，且血清中的TNF-α、IFN-γ、VEGF的含量升高；能显著促进肿瘤组织坏死、消融等现象。水提物组免疫组化则出现负调控，表现出抑瘤率不显著。Western blotting结果表明，与模型组相比各给药组的凋亡基因Caspase-3、Caspase-9和通路基因NF-κB、JAK均高表达，且随剂量的递增表达量逐渐降低。综上所述，杨黄子实体石油醚提取物抑瘤效果显著，其作用机理可能是通过线粒体途径实现，药物经体内代谢后，引起细胞中Bax、Bcl-2、TNF等激活Caspase-3和Caspase-9凋亡蛋白，引起细胞核染色质或DNA凝聚或降解，破坏肿瘤细胞的正常增殖，从而诱导其凋亡，抑制肿瘤的生长。

关键词:瓦尼木层孔菌;细胞凋亡;Caspase-3;Caspase-9;免疫组化染色;Western blotting

Abstract:

In order to explore the antitumor effect and mechanism of different extracts of cultivated Phellinus vaninii fruit body on H₂₂ tumor bearing mice,150 ICR mice were randomly divided into blank group,model group,CTX group,P.vaninii water extract group,ethanol extract group,petroleum ether extract group and crude polysaccharide group.H₂₂ liver cancer cells were used to establish a solid tumor model and the mice were sacrificed on the 10th day after administration.The spleen and thymus organ index and tumor inhibition rate were calculated,the serum levels of TNF-α,INF-γ,VEGF,and hematoxylin-eosin were detected,and the immunohistochemical staining method was used to observe the pathological changes of tumor tissues,while Western blotting was used to detect the expression of tumor-related proteins.The high-dose petroleum ether extract group showed the best tumor inhibition rate (73.21%),increased serum levels of TNF-α,IFN-γ,and VEGF,as well as significantly promoted tumor necrosis and ablation.The immunohistochemistry of the water extract group showed negative regulation,indicating an insignificant tumor suppression.Western blotting showed the apoptosis genes Caspase-3,Caspase-9 and pathway genes NF-κB and JAK were all highly expressed in each administration group compared with the model group,and their expression levels gradually decreased with increasing doses.In summary,the petroleum ether extract of P. vaninii fruit body showed a significant anti-tumor effect which is presumably mediated through the mitochondrial pathway.The metabolism of drug in the body induces activation of Caspase-3 and Caspase-9 apoptotic proteins by Bax,Bcl-2,and TNF,which further caused nuclear chromatin or DNA to condense or degrade,and subsequently destroy the normal proliferation of tumor cells,thereby inducing their apoptosis and inhibiting tumor growth.

Key words:Phellinus vaninii;cytotoxicity;Caspase-3;Caspase-9;immunohistochemistry;Western blotting