灵芝多糖对小鼠胃肿瘤活性的体内外抑制作用

Anti-gastric Tumor Effect of Ganoderma Polysaccharides in Vitro and in Vivo

摘要:

目的：通过体外肿瘤细胞抑制实验及体内抗肿瘤实验，探讨灵芝多糖体内外抗肿瘤作用，并阐明其作用机制。方法：体外实验，使用胃癌细胞株MKN45及AGS，分为空白组、顺铂组、灵芝多糖（20，10，5 g·L^-1）组，加入药物孵育24，48 h后，使用细胞计数试剂盒（cell counting kit-8，CCK-8）检测细胞增殖，流式细胞仪检测细胞周期和凋亡的变化。体内实验，60只BALB/c裸鼠随机分为模型组、阿霉素组、灵芝多糖高、中、低剂量（200，100，50 mg·kg^-1）组，除空白组外，其余各组使用Walker-256肿瘤细胞移植法建立胃肿瘤模型。1周后，除空白和模型组灌胃蒸馏水外，其余各组给予相应药物灌胃。4周后测量肿瘤大小，并取部分胃组织，采用实时荧光定量PCR法（Real-time PCR）检测B淋巴细胞瘤-2（Bcl-2），Bcl-2相关X蛋白（Bax）基因的表达。结果：与空白组比较，灵芝多糖高、中、低剂量组作用24，48 h后，均可以明显抑制胃癌细胞株MKN45和AGS的生长（P<0.05）；灵芝多糖高、中剂量组作用24 h均可以有效促进胃癌细胞MKN45和AGS的凋亡（P<0.05），可以抑制AGS胃癌细胞的细胞周期，使其停留在G₁期（P<0.05）。体内实验表明，灵芝多糖高、中剂量组可以有效抑制胃部肿瘤的生长，同时增加Bax基因表达量，抑制Bcl-2基因表达（P<0.05）。结论：灵芝多糖具有体内外抑制胃肿瘤细胞生长的作用，其作用与增加Bax基因表达，抑制Bcl-2基因表达，从而促进肿瘤细胞凋亡有关。

Abstract:

Objective: To explore the anti-gastric tumor effect and mechanism of Ganoderma polysaccharides based on in vitro and in vivo experiments. Method: In the in vitro experiment, MKN45 and AGS cells were used and randomly divided into blank control, cis-platinum, high, middle and low-dose Ganoderma polysaccharides (20, 10, 5 g·L^-1) groups. Cell counting kit-8 (CCK-8) and flow cytometry methods were used to detect cell proliferation, and flow cytometry was used to detect apoptosis and cycle change, after incubation for 24 and 48 h. In the in vitro experiment, 60 BALB/c naked mice were randomly divided into blank control, model, adriamycin, high, middle and low-dose Ganoderma polysaccharides (200, 100, 50 mg·kg^-1) groups. The gastric tumor model was established by transplanting Walker-256 cell. One week later, each group was given the corresponding drugs. And 4 weeks later, the tumor size was measured, and Real-time PCR method was used to observe the changes in Bcl-2 assaciated X protein (Bax) and B-cell lymphoma-2 (Bcl-2) gene expressions. Result: High, middle and low-dose Ganoderma polysaccharides groups showed inhibition in MKN45 and AGS proliferation in 24, 48 h (P<0.05). High and middle-dose Ganoderma polysaccharides could promote MKN45 and AGS cell apoptosis in 24 h(P<0.05), and inhibit AGS cell cycle in G₁ stage (P<0.05). In vivo experiment showed that high and middle-dose Ganoderma polysaccharides could inhibit the growth of gastric tumor. Meanwhile, the increase on Bax gene expression, and inhibition on Bcl-2 gene expression were observed. Conclusion: Ganoderma polysaccharides could inhibit the growth of gastric tumor in vitro and in vivo. And the mechanism is correlated with improvement on Bax gene expression, inhibition on Bcl-2 gene expression and promotion in tumor cell apoptosis.