黄蘑碱溶性多糖对荷瘤小鼠MMPs表达的调节及其抗肿瘤作用机制

Regulation of huangmo alkali-dissolved polysaccharide on expression of MMPs and its mechanism of anti-tumor effect in tumor-bearing mice

目的:观察黄蘑碱溶性多糖(Fb)对荷瘤小鼠基质金属蛋白酶(MMPs)的调节作用,探讨其抗肝癌作用机理。方法:建立小鼠体内移植性肝癌模型,随机分为5组:阴性对照组(control),阳性对照环磷酰胺组(CTX)和Fb小、中、大剂量治疗组(Fb20,Fb40和Fb80)。测定Fb抑瘤率(IR),观察其整体抗肝癌作用;用明胶酶谱分析和电泳凝胶定量分析系统测定酶解条带的灰度值观察肿瘤组织MMP-2和MMP-9表达水平;免疫组化方法检测种植性肝癌组织内MMPs及其组织抑制因子(TIMPs)的阳性率和分布特点,分析Fb对其蛋白表达的调节作用。结果:Fb20、Fb40和Fb803个治疗组肝癌H22抑瘤率分别为38.7%、51.2%和58.3%,与阴性对照组比较明显增高(P<0.05或P<0.01);Fb40和Fb80组肿瘤组织内TIMP-1表达率分别高达70.0%和84.5%,与阴性对照组(41.7%)和CTX组(44.4%)比较均表现出明显的上调作用(P<0.05);Fb40和Fb80组肿瘤组织内MMP-9的表达分别为47.7%和44.4%,与阴性对照组(60.0%)比较,显示Fb对肝癌组织中MMP-9有下调趋势,但无统计学意义(P>0.05);Fb40组MMP-2表达阳性率为20.0%,与阴性对照(58.3%)比较有明显下调作用(P<0.05)。结论:Fb可以有效地抑制移植性肝癌H22在小鼠体内的生长和侵袭,其作用机制可能与其对MMPs/TIMPs活性的调节有关。

Objective To study the regulation of the huangmo alkali-dissolved polysaccharide （Fb） on expression of MMPs and its anti-hepatocarcinama effect in tumor-bearing mice. Methods Transplanted liver cancer models of mice were set up, the mice were equally divided into control group, CTX group, and three treatment groups of low, middle, high doses of Fb （Fb20, Fb40 and Fb80）. The inhibitory rate （IR） was determined to observe the anti-tumor effect in vivo ; Gelatin zymography and quantified analytical system were used to detect gray-scale value of straps to observe the expressions of MMP-2 and MMP-9; the regulation of Fb on the protein expressions of MMP-2, 9 and TIMP-1 was analyzed, the positive rate was detected by immunohistochemistry. Results IR of Fb20, Fb40 and Fb80 groups （38.70%, 51.2% and 58.3%） were higher than that in control group （P〈0.05 or P〈0.01）; the expression rates of TIMP-1 in Fb40 and Fb80 groups （70.0% and 84.5%） were higher than those in control group and CTX group （41.7% and 44. 4%） （P〈 0. 05）, the expressions of MMP-9 were down-regulated in Fb40 （47.7%） and Fb80 （44. 4%） groups, but there was no difference, compared with control group （60.0%） （P〉 0. 05）. The expression of MMP-2 in Fb40 group （20.0%） was lower than that in control group （58.3%） （P〈 0. 05）. Conclusion Fb has strong activity to restrain the tumor growth and metastasis in transplanted liver cancer H22, the anti-tumor mechanism of Fb probably correlates with its effect on regulating the activities of protein expressions of MMPs and TIMPs.