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目的:探讨益气除痰方对BALB/c裸小鼠肺癌A549移植瘤的抑制作用及其机制。方法:人肺腺癌细胞A549接种BALB/c裸鼠40只,随机分为模型组(生理盐水)、顺铂注射液(0.002 g/kg)组、益气除痰方低剂量(3.0g/kg)组、益气除痰方高剂量(6.0 g/kg)组、联合用药(益气除痰方6.0 g/kg+顺铂注射液组0.002 g/kg)组。造模第8天,中药灌胃,1次/d,每次0.2 m L,连续14 d。第17天,顺铂注射液腹腔注射,1次/d,每次0.2 m L,连续5 d。第22天,麻醉处死裸鼠,检测瘤体积和瘤质量,计算抑瘤率。采用免疫组织化学法、Western blot法及RTQ-PCR法检测肿瘤组织中钙联蛋白(calnexin,CNX)及调控转录因子X盒结合蛋白1(X-box binding protein 1,XBP1)的表达。结果:顺铂注射液组、益气除痰方高剂量组及联合用药组瘤质量及瘤体积较模型组显著降低(P<0.05或P<0.01),且联合用药组显著优于顺铂注射液组(P<0.01)。免疫组织化学法、Western blot法及RTQ-PCR法结果均显示顺铂注射液组、益气除痰方高剂量及联合用药组CNX及XBP1的表达较模型组显著降低(P<0.01),且联合用药组优于顺铂注射液组(P<0.05或P<0.01)。结论:益气除痰方能抑制A549肺癌生长,与顺铂联用能起到增效作用,其机制可能与下调相关分子伴侣蛋白CNX及XBP1的表达而抑制未折叠蛋白反应(unfolded protein response,UPR),从而导致肿瘤细胞凋亡有关。
Objective: To investigate the inhibitory effect of Yiqi Chu Tan Recipe on lung cancer A549 xenografts in BALB / c nude mice and its mechanism. Methods: Forty BALB / c nude mice were inoculated with human lung adenocarcinoma A549 cells and randomly divided into model group (normal saline), cisplatin injection (0.002 g / kg), low dose of Yiqi Chu Tan (3.0g / kg), and the combination of Yiqi Chu Tan Recipe (6.0 g / kg) and Yiqi Chu Tan Recipe (6.0 g / kg + cisplatin 0.002 g / kg). On the 8th day of modeling, traditional Chinese medicine was given intragastrically once daily for 0.2 m L for 14 days. On the 17th day, cisplatin injection was injected intraperitoneally once a day for 0.2 days for 5 days. On the 22nd day, the nude mice were killed by anesthesia, the tumor volume and tumor mass were measured, and the tumor inhibition rate was calculated. Immunohistochemistry, Western blot and RT-PCR were used to detect the expression of calnexin (CNX) and XBP1. Results: The tumor mass and tumor volume of cisplatin injection group, high-dose Yiqizhitan formula group and combination group were significantly lower than those of model group (P <0.05 or P <0.01), and the combined treatment group was significantly better than cisplatin injection Liquid group (P <0.01). The results of immunohistochemistry, Western blot and RTQ-PCR showed that the expressions of CNX and XBP1 in Cisplatin injection group, Yiqi Zutan Recipe high dosage group and combination group were significantly lower than those in model group (P <0.01), and Combination group than cisplatin injection group (P <0.05 or P <0.01). CONCLUSION: Yiqi Shutan can inhibit the growth of A549 lung cancer and synergize with cisplatin. The mechanism may be related to the down-regulation of the expression of the related chaperones CNX and XBP1 and the unfolded protein response UPR), leading to tumor cell apoptosis.