益气养阴方抗C57小鼠Lewis肺癌转移作用与调节MMP-9、TIMP-1蛋白表达的实验研究

Experimental Research on Anti-metastasis Effect of "Yiqi Yangyin Decoction" and Its Regulatory Effects on Expressions of MMP-9 and TIMP-1 of Lewis Lung Cancer in C57 Mice

目的免疫组化法观察益气养阴方对C57BL/6小鼠Lewis肺癌瘤体MMP-9、TIMP-1蛋白表达的影响以及抗肺转移作用,探讨该方抗转移作用的机理。方法24只C57BL/6小鼠随机分为3组,每组8只,接种Lewis肺癌瘤株,造模后第2天按如下方案进行干预:荷瘤对照组:0.9%NaCl溶液灌服;中药组:益气养阴方煎汤灌服;顺铂组:顺铂溶液腹腔内注射。第20天处死全部小鼠,称瘤重计算抑瘤率,计数肺转移数,计算抑制肺转移率,免疫组化Envision二步法测定瘤体MMP-9、TIMP-1蛋白表达情况。结果中药组和顺铂组的抑瘤率分别为35.02%和41.18%,具有统计学意义(P<0.05)。中药组和顺铂组的抑制肺转移率分别为54.20%和39.02%,差异无统计学意义。瘤重与肺转移数之间存在正相关。荷瘤对照组MMP-9和TIMP-1阳性率均为100%,中药组MMP-9阳性率显著低于对照组,TIMP-1阳性表达与对照组比较差异无统计学意义,但中度及强阳性率明显低于对照组。MMP-9和TIMP-1的表达与肺转移数成正相关,两者之间亦存在正相关。结论益气养阴方具有较好的抑瘤作用和抑制肺转移的作用;肺癌均表达MMP-9和TIMP-1蛋白,是促进转移的重要机理和判断指标;益气养阴方能够显著下调肺癌MMP-9的表达,影响TIMP-1的表达,这是其抑制肺癌转移的机理之一。

Objective To observe the effect of “Yiqi Yangyin Decoction” on the expressions of MMP-9 and TIMP-1 of Lewis lung cancer in C57BL/6 mice with immunohistechemistry and its anti-metastasis mechanisms. Methods Twenty-four C57BL/6 mice were randomized into three groups and all transplanted with Lewis lung cancer cells, and then received following interventions next day： Tumor-bearing control group was fed with 0.9% NaC1, and TCM group was treated with this Decoction, and DDP group was given intraperitaneal DDP injection. All of mice were sacrificed on the 20th day, weighed the tumor and counted lung metastases to calculate the inhibition rates of tumor growth and metastasis. The expressions of MMP-9, TIMP-1 in tumor tissues were detected by immunohistochemistry. Results The inhibition rates of tumor growth and metastasis were 35.02% and 54.20% in TCM group, and 41.18% and 39.02% in DDP group respectively（P 〈 0.05）. Tumor weight was positively correlated to lung metastasis. The positive rates of MMP-9 and TIMP-1 were both 100% in tumor-bearing group; the expres- sion rate of MMP-9 in TCM group was significantly lower than that in control group. Although there was no obvious difference of the expression rate of TIMP-1 between TCM group and control group, the moderate and strong positive rates in TCM group were obviously lower than in control group. The expressions of MMP-9 and TIMP-1 were both positively correlated to lung metastasis, and there was a positive correlation between these two proteins. Conclusion This Decoction has good effects on tumor-inhibition and anti-metastasis by down-regulating MMP-9 and TIMP-1 expressions.