首页|黄芪-莪术对Lewis肺癌小鼠NF-κB通路及肿瘤自噬的干预作用研究

黄芪-莪术对Lewis肺癌小鼠NF-κB通路及肿瘤自噬的干预作用研究

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目的 探讨黄芪-莪术配伍抑制NF-κB通路及诱导Lewis肺癌小鼠肿瘤自噬的作用机制.方法 采用腋窝皮下接种肺癌Lewis细胞悬液法建立Lewis肺癌小鼠模型,建模成功后将40只小鼠按随机数字表法分为模型组、中药组、顺铂组和中药+顺铂组,每组各10只.在建模后第3天开始给药,顺铂组给予腹腔注射顺铂(2 mg/kg),1次/3 d,共6次,并每天予灭菌双蒸水灌胃(8.2 g/kg);中药组每日予黄芪-莪术浓煎液灌胃(8.2 g/kg),1次/d,连续18 d,并腹腔注射稀释液[2 mg/(kg·3 d)];中药+顺铂组按照以上给药方法予中药和顺铂;模型组每日予等体积灭菌双蒸水灌胃并腹腔注射稀释液剂量同上.期间观察小鼠毛发、动态及进食、饮水情况,每2 d测量小鼠体质量、每4 d测算小鼠肿瘤体积.在造模20 d后,Western Blot法检测肿瘤组织NF-KBp65、p-NF-κB p65、IκB-α、p-IκB-α、LC3和Beclin-1的蛋白表达水平,ELISA法测定肿瘤组织中TNF-α和IL-1β的含量,并采用透射电镜观察肿瘤细胞超微形态的变化.结果 与模型组比较,中药组体质量无明显差异(P>0.05),顺铂组、中药+顺铂组体质量明显较轻,差异有统计学意义(P<0.01),各用药组小鼠肿瘤体积明显减小,差异有统计学意义(P<0.01)、LC3-Ⅱ/LC3-Ⅰ及Beclin-1/β-actin值均明显升高,差异有统计学意义(P<0.01)、p-NF-κB p65/NF-κB p65及pIκBα/IκBα值、TNF-α和IL-1β均明显降低,差异有统计学意义(P<0.05);与顺铂组比较,中药+顺铂组肿瘤体积值无明显差异(P>0.05)、Beclin-1/β-actin值明显升高,差异有统计学意义(P<0.01)、p-NF-κB p65/NF-KB p65及pIκBα/IκBα值及TNF-α和IL-1β均明显降低,差异有统计学意义(P<0.05),LC3-Ⅱ/LC3-Ⅰ值无明显差异(P>0.05);与中药组比较,中药+顺铂组体质量较轻(P<0.01),肿瘤体积较小,差异有统计学意义(P<0.01),Beclin-1/β-actin值升高,差异有统计学意义(P<0.01),p-NF-κB p65/NF-κB p65、pIκBα/IκBα、TNF-α 和 IL-1 β 值均降低,差异有统计学意义(P<0.05),LC3-Ⅱ/LC3-Ⅰ 值差异无显著性意义(P>0.05).结论 黄芪-莪术配伍可以通过抑制IκB-α的磷酸化减少其降解,达到抑制NF-κB活化,并改善肿瘤炎性微环境,同时上调自噬相关蛋白的表达水平,诱导细胞自噬,进而抑制肿瘤生长.
Impact of Radix Astragali Rhizoma Curcumae on the NF-κB Pathway and Tumor Autophagy in Lewis Lung Cancer Mice
Objective To investigate the mechanism of action of Radix Astragali-Rhizoma Curcumae in inhibi-ting the NF-κB pathway and inducing tumor autophagy in Lewis lung cancer mice.Methods The Lewis lung cancer mouse model was established through subcutaneous injection of Lewis cell suspension into the axillary region.After suc-cessful modeling,40 mice were assigned into four groups by random number table:model group,Chinese medicine group,cisplatin group,and Chinese medicine+cisplatin group,with 10 in each group.On the 3rd day after modelling,the cisplatin group received intraperitoneal injection of cisplatin(2 mg/kg),once every three days for a total of six doses.Additionally,they were administered sterilized double-distilled water by gavage(8.2 g/kg)daily.The Chinese medicine group received Radix Astragali Rhizoma Curcumae concentrated decoction(8.2 g/kg)by gavage,once per day for 18 consecutive days,a-long with intraperitoneal injection of diluted solution(2 mg/kg·3d).The Chinese medicine+cisplatin group was given both Chinese medicine and cisplatin following the same administration method mentioned above.For the model group,an e-quivalent volume of sterilized double-distilled water was administered by gavage,and the same diluted solution was ad-ministered via intraperitoneal injection on a daily basis.Throughout the study period,the mice were observed for changes in hair,activity,food consumption,and water intake.The body mass of the mice was measured every two days,and the tumor volume was measured every four days.After 20 days of tumor modeling,the protein expressions of NF-κB p65,p-NF-κB p65,IκB-α,p-IκB-α,LC3,and Beclin-1 in the tumor tissues were assessed through Western Blot.Additionally,the levels of TNF-α and IL-1 β in the tumor tissues were determined using ELISA.The ultramorphological changes of tumor cells were examined by transmission electron microscopy.Results Compared with the model group,the Chinese medicine group showed no significant difference in body mass(P>0.05),while the cisplatin and Chinese medicine+cis-platin groups exhibited significantly lower body mass(P<0.01).The tumor volume was reduced(P<0.01),and LC3-Ⅱ/LC3-Ⅰ and Beclin-1/β-actin were up-regulated in all treatment groups(P<0.01),while p-NF-κB p65/NF-κB p65,pIκBα/IκBα,TNF-α,and IL-1β were down-regulated(P<0.05).Compared with the cisplatin group,the Chinese medicine+cisplatin group had no obvious difference in tumor volume and LC3-Ⅱ/LC3-Ⅰ(P>0.05),but had increased Beclin-1/β-actin(P<0.01)and decreased p-NF-κB p65/NF-κB p65,pIκBα/IκBα,TNF-α,and IL-1β(P<0.05).The Chinese medicine+cisplatin group had lower body mass(P<0.01),smaller tumor volume(P<0.01),higher Beclin-1/β-actin(P<0.01),and lower p-NF-κB p65/NF-κB p65,pIκBα/IκBα,TNF-α,and IL-1β(P<0.05)than the Chinese medicine group.The difference in LC3-Ⅱ/LC3-Ⅰ was not significant between the two groups(P>0.05).Conclusion Radix Astragali Rhizoma Curcumae has the ability to inhibit the activation of NF-κB and improve the inflammatory microenvironment of tumors.This is achieved by reducing the degradation of IκB-α through the inhibition of its phosphorylation.Moreover,the combination up-regulates the expression of autophagy-related proteins and induces cellular autophagy,ultimately leading to the inhibition of tumor growth.

Radix Astragali Rhizoma CurcumaeNF-κBAutophagyTumor

于泓洋、李潇、田培裕、窦永起

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中国人民解放军医学院,北京 100853

71901部队,山东聊城 252000

中国人民解放军总医院第六医学中心中医医学部,北京 100089

黄芪-莪术 NF-κB 自噬 肿瘤

国家自然科学基金资助项目

82174118

2024

世界中西医结合杂志
中华中医药学会

世界中西医结合杂志

CSTPCD
影响因子:1.053
ISSN:1673-6613
年,卷(期):2024.19(1)
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