| 杨雪飞,张志娣,黄挺,傅晓青,叶知锋,郭俊华,刘冰.益气养阴散结方对A549裸小鼠肿瘤脂滴相关蛋白与脂代谢相关蛋白的影响[J].浙江中西医结合杂志,2023,33(3): |
| 益气养阴散结方对A549裸小鼠肿瘤脂滴相关蛋白与脂代谢相关蛋白的影响 |
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| 投稿时间：2022-06-27 修订日期：2023-02-23 |
| DOI： |
| 中文关键词: 非小细胞肺癌 人肺腺癌细胞株A549 益气养阴散结方 脂滴相关蛋白 脂代谢相关蛋白 |
| 英文关键词:non-small cell lung cancer human alveolar epithelial cell line A549 Yiqi Yangyin Sanjie Decoction lipid droplet-associated proteins lipid metabolism-associated proteins |
| 基金项目:1.项目名称：益气养阴散结方对非小细胞肺癌模型小鼠肿瘤细胞脂滴异常的影响，资助部门：浙江省基础公益研究计划项目，项目编号：LGF18H290001； 2.项目名称：益气养阴散结方干预非小细胞肺癌肿瘤细胞脂滴异常的机制研究，资助部门：浙江省卫生计生委中医药管理局，项目编号：2020ZB160； 3. 张志娣全国名老中医药专家传承工作室。 |
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| 中文摘要: |
| 摘要 目的 考察益气养阴散结方对A549裸小鼠肿瘤脂滴相关蛋白、脂代谢相关蛋白的影响。方法取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5组，每组6只，即：模型组，顺铂组和益气养阴散结方低、中、高剂量组。接种次日起给药，模型组给予生理盐水0.3 ml，顺铂组给予顺铂4mg/kg，益气养阴散结方低、中、高剂量组5g/kg 、10g/kg、20g/kg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（lipid storage droplet protein 5，LSDP5）、47KDa尾连蛋白（tail interacting protein of 47 kDa，TIP47）、脂肪分化相关蛋白（Adipose Differentiation-Related Protein，ADRP）、脂肪细胞型脂肪酸结合蛋白（adipocyte-fatty acid binding protein，A-FABP）、Delta样因子-1（Delta-like 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 免疫印迹结果表明，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，差异具有统计学意义（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，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，差异具有统计学意义（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；A-FABP，4.02±0.40、3.40±0.23比5.67±0.75；DLK1，5.10±0.40比7.93±0.91；caveolin-1，5.03±0.50、4.23±0.35比6.33±0.93。P＜0.05或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、脂代谢相关蛋白在肿瘤中的表达。 |
| 英文摘要: |
| Abstract Objective To explore effects of Yiqi Yangyin Sanjie Decoction on the expression of lipid droplet-associated proteins in A549 tumor-bearing nude mice. Methods A total of 30 male BALB/c mice aged from 4 to 6 weeks were subcutaneously inoculated with A549 human lung cancer cell line via axilla to establish tumor-bearing nude mice models, and randomly divided into five groups: model group, cisplatin group, Yiqi Yangyin Sanjie(YQYYSJ) Decoction low/medium/high dose groups.From the next day of tumor inoculation, model group were given normal saline 0.3ml/d,cisplatin group were given cisplatin 4mg/kg?d,YQYYSJ Decoction low/medium/high dose groups were given YQYYSJ Decoction 5g/kg?d, 10g/kg?d,and 20g/kg?d for 8 weeks. To examine the expression 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（A-FABP）, Delta-like 1（DLK-1）and Caveolin-1, western blotting and immunohistochemical analyses were performed. Results Western blotting analysis showed the expression of many a lipid droplet- and lipid metabolism-associated proteins significantly decreased in low and high dose groups of YQYYSJ Decoction, compared with model group(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；FABP, 0.52±0.14 and 0.31±0.09 vs. 0.59±0.19；DLK1, 0.75±0.23 and 0.64±0.08 vs. 1.07±0.21；caveolin-1, 0.60±0.25 and 0.41±0.09 vs. 1.09±0.31. P＜0.05 or P＜0.01). Immunohistochemical analysis revealed immunohistochemical scores of many a lipid droplet- and lipid metabolism-associated proteins significantly decreased in medium and high dose groups of YQYYSJ Decoction, compared with model group(Perilipin-1, 4.17±0.32 and 3.90±0.57 vs.5.70±0.97；TIP47, 5.13±0.50 and 3.73±0.31 vs.5.67±0.70；ADRP, 4.67±1.01and 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；DLK1, 5.10±0.40 vs.7.93±0.91；caveolin-1, 5.03±0.50 and 4.23±0.35 vs.6.33±0.93; P＜0.05 or P＜0.01). Conclusion High dose of YQYYSJ Decoction can significantly decreased the expression of lipid droplet- associated proteins, suggesting a promising future of experimental and clinical studies. |
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