化痰散结方对肿瘤间质纤维母细胞活化相关蛋白表达的影响

Effect of Huatan Sanjie Recipe on Fibroblast Activition Protein in Orthotopic Transplanted Tumor in Rat Model

目的探讨化痰散结方对肿瘤间质纤维母细胞活化状态的影响。方法 walker-256细胞株建立大鼠胃种植瘤模型,40只大鼠,36只于胃部接种瘤块,其余4只不接种的大鼠为正常对照组,种植瘤块的Wistar大鼠随机分为化痰散结方高剂量组,化痰散结中药3m L/只(含生药3g·m L-1),化痰散结方低剂量组,化痰散结中药3m L/只(含生药1g·m L-1),模型对照组(给予生理盐水3m L/只);每组12只,各组于造模后次日立即给药,每日灌胃,连续给药21d;正常对照组每日仅正常饮食饲养;21d后通过免疫组化、Western-blot、Real-time PCR法检测各组胃种植瘤组织中及正常大鼠胃组织中CD34、成纤维细胞激活蛋白(fibroblast activation protein alpha,FAPα)、平滑肌肌动蛋白(α-smooth muscle actin,α-Sma)的表达。结果模型对照组与化痰散结方高、低剂量组胃种植瘤中CD34的蛋白及基因表达水平明显低于正常对照组大鼠胃组织(P<0.05或<0.01),而其FAPα和α-SMA的表达则明显高于正常对照组(P<0.05);与模型对照组相比,中药高、低剂量组CD34的表达均升高(P<0.05或<0.01);而中药高剂量组中FAPα与α-SMA的表达则明显低于模型对照组(P<0.01)。结论化痰散结方可以降低肿瘤间质纤维母细胞活化相关蛋白的表达,对间质纤维母细胞活化有一定的阻断作用。

Objective To investigate effect of Huatan Sanjie recipe on fibroblast activated status in tumor growth. Methods Walker- 256 cell lines were taken as the origin of rats model. Orthotopic transplanted tumor in rat model was set up 36 rats model and they were randomly divided into Chinese medicine high dose group,Chinese medicine low dose group,and model group. At the same time,4 rats were used as normal control group. The rats of Chinese medicine group were given Huatan Sanjie recipe. Rats in the model group were given normal saline while rats of normal control group wasn't intervened. All rats raised under the same conditions. After 21 days,the tumor tissue and normal gastric tissue were removed. Expression of CD34,fibroblast activationprotein alpha( FAPα) and alpha smooth muscle actin( α- Sma) in these tissue were observed by using immunohistochemical technique,Western- blot and Real- time PCR. Results Expression of CD34 in normal group was significant higher than that of the other three groups( P < 0. 05 or < 0. 01). However,expression of FAPα and α- Sma in this group was significant lower than those of the others( P < 0. 05 or < 0. 01).Compared with model group,Expression of CD34 in Chinese medicine group increased significantly( P < 0. 05 or < 0. 01),but expression of FAPα and α- Sma in Chinese medicine high dose group was significantly lower than that of model group( P < 0. 01). Conclusion Huatan Sanjie recipe can reduce the expression of fibroblast activation related proteins.