【摘要】 目的 探讨益气养阴散结方对A549裸鼠肿瘤脂滴相关蛋白、脂代谢相关蛋白的影响。方法取4～6周龄BALB/c裸鼠30只，皮下接种A549细胞株以复制肺腺癌小鼠模型，按照随机数字表法分为五组，模型组、顺铂组和益气养阴散结方低、中、高剂量组，每组6只。接种次日起给药，模型组给予生理盐水0.3 mL/d；顺铂组给予顺铂4 mg/kg，每周2次；益气养阴散结方低、中、高剂量组给予5、10、20 g/（kg·d）益气养阴散结方，连续给药8周。采用免疫印迹、免疫组化检测脂周素-1(perilipin-1)、脂滴蛋白5(LSDP5)、47kDa尾连蛋白（TIP47）、脂肪分化相关蛋白（ADRP）、脂肪细胞型脂肪酸结合蛋白（A-FABP）、Delta样因子-1(DLK-1)、小窝蛋白-1(caveolin-1)等脂滴相关蛋白、脂代谢相关蛋白在肿瘤中的表达。结果 30只裸鼠成瘤，成瘤率100%。在实验期内，模型组和益气养阴散结方中剂量组各死亡裸鼠1只。免疫印迹结果表明，与模型组比较，益气养阴散结方低、高剂量组显著下调脂滴相关蛋白、脂代谢相关蛋白的表达[perilipin-1:(0.76±0.23)、（0.58±0.18）比（0.93±0.22）;LSDP5:(0.79±0.23)、（0.38±0.11）比（1.06±0.30）;TIP47:(0.49±0.19)、（0.24±0.06）比（0.71±0.25）;ADRP:(0.48±0.15)、（0.27±0.08）比（0.61±0.17）;A-FABP:(0.52±0.14)、（0.31±0.09）比（0.59±0.19）;DLK1:(0.75±0.23)、（0.64±0.08）比（1.07±0.21）;caveolin-1:(0.60±0.25)、（0.41±0.09）比（1.09±0.31）,P<0.05或P<0.01]。免疫组化结果表明，与模型组比较，益气养阴散结方中、高剂量组显著下调脂滴相关蛋白、脂代谢相关蛋白的表达[perilipin-1:(4.17±0.32)、（3.90±0.57）比（5.70±0.97）;TIP47:(5.13±0.50)、（3.73±0.31）比（5.67±0.70）;ADRP:(4.67±1.01)、（4.17±0.61）比（6.13±0.81）;AFABP:(4.02±0.40)、（3.40±0.23）比（5.67±0.75）;caveolin-1:(5.03±0.50)、（4.23±0.35）比（6.33±0.93）,P<0.05或P<0.01]，高剂量组下调DLK1表达[（5.10±0.40）比（7.93±0.91）,P<0.01]。结论 高剂量益气养阴散结方可显著降低perilipin-1、LSDP5、TIP47、ADRP、A-FABP、DLK1、caveolin-1等脂滴相关蛋白、脂代谢相关蛋白在A549裸鼠肿瘤中的表达。更多还原

【Abstract】 Objective To assess effects of the Yiqi Yangyin Sanjie(YQYYSJ) recipe on expression of lipid droplet-and lipid metabolism-associated proteins in nude mouse lung cancer A549 cell xenografts. Methods A total of 30 male BALB/c mice of 4 to 6 weeks old were subcutaneously inoculated with human lung cancer A549cells and then randomly divided into model, cisplatin, and YQYYSJ at low, medium, and high dose(n =6 per group). One day after tumor cell injection, the model mice were given normal saline(0.3 mL/d) and cisplatin-treated mice received 4 mg/kg cisplatin twice a week, while mice treated with the YQYYSJ recipe were given YQYYSJ at 5, 10, and 20 g/（kg·d）, respectively for 8 weeks. At the end of experiments, mice were sacrificed and tumor xenografts were resected for assessment of perilipin-1, lipid storage droplet protein 5(LSDP5), tail interacting protein of 47 kDa(TIP47), adipose differentiation-related protein(ADRP), adipocyte-fatty acid binding protein(AFABP), Delta-like 1(DLK-1), and caveolin-1 using Western blot and immunohistochemistry, respectively. Results There was 100% tumorigenesis rate in these 30 nude mice. However, during the experimental period of time, one mouse each was dead in the model and medium dose of YQYYSJ treatment groups. However, low and high doses of YQYYSJ treatment significantly reduced expression of various lipid droplet-and lipid metabolism-associated proteins compared with those of the model group(P<0.05 or P<0.01), i.e., perilipin-1(0.76±0.23 and 0.58±0.18 vs.0.93±0.22), LSDP5(0.79±0.23 and 0.38±0.11 vs. 1.06±0.30, TIP47(0.49±0.19 and 0.24±0.06 vs. 0.71±0.25), ADRP(0.48±0.15 and 0.27±0.08 vs. 0.61±0.17), A-FABP(0.52±0.14 and 0.31±0.09 vs. 0.59±0.19), DLK1(0.75±0.23 and0.64±0.08 vs. 1.07±0.21), and caveolin-1(0.60±0.25 and 0.41±0.09 vs. 1.09±0.31). Moreover, immunohistochemical analysis also confirmed medium and high doses of YQYYSJ treatment in reduction of these proteins in mouse xenografts(P<0.05 or P<0.01), i.e., perilipin-1(4.17±0.32 and 3.90±0.57 vs. 5.70±0.97), TIP47(5.13±0.50 and3.73±0.31 vs. 5.67±0.70), ADRP(4.67±1.01 and 4.17±0.61 vs. 6.13±0.81), A-FABP(4.02±0.40 and 3.40±0.23 vs.5.67±0.75), caveolin-1(5.03±0.50 and 4.23±0.35 vs. 6.33±0.93), while a high YQYYSJ dose downregulated DLK1level, compared to that of the model group(5.10±0.40 vs. 7.93±0.91; P<0.01). Conclusion A high YQYYSJ dose was able to significantly reduce level of lipid droplet-associated proteins(perilipin-1, LSDP5, TIP47, ADRP, AFABP, DLK1 and caveolin-1)in lung cancer A549 cell xenografts in nude mice.更多还原