养阴扶正汤调控肺腺癌Warburg 效应分子机制研究
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R285.5

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黑龙江自然科学基金项目(H2016095)


Study on Molecular Mechanism of Yangyin Fuzheng Tang Regulating Warburg Effect of Lung Adenocarcinoma
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    摘要:

    目的:探讨养阴扶正汤调控肺腺癌Warburg 效应的分子机制。方法:建立人肺腺癌A549 裸鼠移植瘤模型,模型制备成功后,随机分为模型组(Control 组),养阴扶正汤低、中、高剂量组(YYFZL 组、YYFZM 组、YYFZH 组),顺铂组(DDP组),顺铂联合养阴扶正汤低、中、高剂量组(DDP+YYFZL 组、DDP+YYFZM 组、DDP+YYFZH 组) 8 组,每组6 只。除Control组外,各组用相应的药物干预14 天后处死,绘制肿瘤生长曲线、称取瘤重;检测瘤组织中乳酸、三磷酸腺苷(ATP) 的含量;Western blot、RT-PCR 检测瘤组织中葡萄糖转运体1 (GLUT1)、已糖激酶2 (HK2)、丙酮酸激酶(PKM)、乳酸脱氢酶A(LDHA) 的蛋白质及mRNA 表达。结果:与Control 组比较,其余各组移植瘤体积均减小(P<0.01);与DDP 组比较,DDP+YYFZH 组A549 移植瘤体积减小(P<0.05)。与Control 组比较,其余各组瘤重降低(P<0.01,P<0.05);且随着养阴扶正汤剂量的升高,瘤重下降;与DDP 组比较,DDP+YYFZL 组、DDP+YYFZM 组、DDP+YYFZH 组瘤重降低(P<0.05)。与Control 组比较,除YYFZL 组外,其余各组瘤组织乳酸含量减少(P<0.01);与DDP 组比较,DDP+YYFZH 组瘤组织乳酸含量减少(P<0.01)。与Control 组比较,YYFZM 组、YYFZH 组、DDP 组、DDP+YYFZM 组、DDP+YYFZH 组瘤组织ATP 含量减少(P<0.01)。与Control 组比较,其余各组HK2、LDHA 蛋白相对表达量减少(P<0.01,P<0.05);除YYFZL 组以外,其余各组GLUT1 蛋白相对表达量减少(P<0.01,P<0.05);除YYFZL 组外,其余各组PKM 蛋白相对表达量减少(P<0.01,P<0.05);与DDP 组比较,DDP+YYFZH 组LDHA 蛋白相对表达量减少(P<0.05)。结论:养阴扶正汤可通过降低肺腺癌Warburg 效应代谢途径中关键酶的蛋白质表达,来抑制Warburg 效应,从而起抗癌作用。

    Abstract:

    Abstract:Objective:To investigate the molecular mechanism of Yangyin Fuzheng tang regulating Warburg effect of lung adenocarcinoma. Methods:After the successful establishment of human lung adenocarcinoma A549 nude-mice transplanted tumor models, they were randomly divided into eight groups: the model group(the control group), the Yangyin Fuzheng tang groups with low,medium and high dose respectively(the YYFZL group,YYFZM group and YYFZH group),the cisplatin group(the DDP group), the cisplatin and Yangyin Fuzheng tang groups with low, medium and high dose respectively(the DDP + YYFZL group, DDP + YYFZM group and DDP + YYFZH group), with six mice in each group. Except for those in the control group, the mice in the other groups were sacrificed after 14 days of corresponding drug intervention; the tumor growth curve was drawn,and the tumor was weighed;the contents of lactic acid and adenosine triphosphate(ATP) in tumor tissues were detected;the protein and mRNA expressions of glucose transporter1(GLUT1),hexokinase 2(HK2),pyruvate kinase(PKM) and lactate dehydrogenase A(LDHA) in tumor tissues were detected by Western blot and RT- PCR. Results: Compared with that in the control group, the volumes of transplanted tumor in other groups were all decreased(P<0.01); compared with that in the DDP group,the volume of transplanted tumor A549 in the DDP+YYFZH group was decreased(P< 0.05). Compared with that in the control group,the tumor weight in the other groups were all decreased(P<0.01,P<0.05); with the increase of the dose of Yangyin Fuzheng tang,the tumor weight was decreased;compared with that in the DDP group,the tumor weight in the DDP+YYFZL group,the DDP+YYFZM group and the DDP+YYFZH group were decreased (P<0.05). Compared with that in the control group,the contents of lactic acid in tumor tissues in the other groups except for the YYFZL group were decreased(P<0.01);compared with that in the DDP group,the content of lactic acid in tumor tissues in the DDP + YYFZH group was decreased(P<0.01). Compared with that in the control group, ATP contents in tumor tissues in the YYFZM group, the YYFZH group, the DDP group, the DDP + YYFZM group and the DDP + YYFZH group were decreased(P<0.01). Compared with that in the control group, the relative protein expression of HK2 and LDHA in the other groups were decreased(P<0.01,P<0.05);relative protein expressions of GLUT1 in the other groups except for the DDP+YYFZL group were decreased(P<0.01,P<0.05);except for that in the YYFZL group,relative protein expressions of PKM in the other groups were decreased(P<0.01,P<0.05). Compared with that in the DDP group,the relative expression of LDHA in the DDP + YYFZH group was decreased(P<0.05). Conclusion: Yangyin Fuzheng tang can inhibit the Warburg effect by reducing the protein expressions of key enzymes in the metabolic pathway of Warburg effect in lung adenocarcinoma,thus playing an anticancer role.

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陈宏,孙玺媛,尹钢,魏冬梅,姜梅,刘锐.养阴扶正汤调控肺腺癌Warburg 效应分子机制研究[J].新中医,2020,52(20):1-5

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  • 在线发布日期: 2020-10-19
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