江西中医药大学
纸质出版:2021
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[1]李佳萍,余功,谢斌.清燥救肺汤对Lewis肺癌荷瘤小鼠肺癌组织氧化磷酸化能量代谢的影响[J].中医杂志,2021,62(05):439-444.
LI Jiaping, YU Gong, XIE Bin. Effect of Qingzao Jiufei Decoction (清燥救肺汤) on Energy Metabolism of Oxidative Phosphorylation in Lung Carcinoma Tissue of Lewis Tumor-Bearing Mice[J]. Journal of Traditional Chinese Medicine2021, 62(5): 439-444.
[1]李佳萍,余功,谢斌.清燥救肺汤对Lewis肺癌荷瘤小鼠肺癌组织氧化磷酸化能量代谢的影响[J].中医杂志,2021,62(05):439-444. DOI: 10.13288/j.11-2166/r.2021.05.015.
LI Jiaping, YU Gong, XIE Bin. Effect of Qingzao Jiufei Decoction (清燥救肺汤) on Energy Metabolism of Oxidative Phosphorylation in Lung Carcinoma Tissue of Lewis Tumor-Bearing Mice[J]. Journal of Traditional Chinese Medicine2021, 62(5): 439-444. DOI: 10.13288/j.11-2166/r.2021.05.015.
目的探讨清燥救肺汤治疗肺癌的可能作用机制。方法 50只C57BL/6J小鼠随机分为模型组
环磷酰胺(CTX)组
清燥救肺汤高、中、低剂量组
每组10只。各组小鼠分别接种Lewis肺癌细胞建立肺癌荷瘤小鼠模型。24h后
模型组每日给予0.2ml生理盐水灌胃
每日2次;CTX组隔日1次以50mg/kg CTX腹腔注射;清燥救肺汤高、中、低剂量组分别以11、5.5、2.75g/(kg·d)清燥救肺汤灌胃
每日2次。清燥救肺汤各剂量组在接种前后14天均灌胃
其余各组在接种细胞24h后继续给药14天。Real-time PCR法检测肺癌组织中三磷酸腺苷合酶β亚基(ATP 5B)mRNA表达
蛋白免疫印迹法检测细胞色素C氧化酶亚单位Ⅳ(COXⅣ)蛋白表达
高效液相色谱法检测一磷酸腺苷(AMP)、二磷酸腺苷(ADP)、三磷酸腺苷(ATP)的含量;JC-1法检测线粒体膜电位(MMP)变化。结果与模型组比较
CTX组及清燥救肺汤高、中剂量组ATP 5B mRNA
COXⅣ蛋白表达、ATP含量和线粒体膜电位显著降低(P<0.05或P<0.01);与CTX组比较
清燥救肺汤高、中、低剂量组ATP 5B mRNA、COXⅣ蛋白表达升高
清燥救肺汤中、低剂量组ATP含量、MMP水平
清燥救肺汤高剂量组AMP/ATP显著升高
清燥救肺汤中、低剂量组AMP/ATP及清燥救肺汤低剂量组ADP/ATP显著降低(P<0.05或P<0.01);与清燥救肺汤高剂量组比较
清燥救肺汤中、低剂量组ATP 5B mRNA、COXⅣ蛋白表达、ATP含量升高
AMP/ATP值、ADP/ATP值显著降低
清燥救肺汤低剂量组MMP水平明显升高(P<0.05或P<0.01)。与清燥救肺汤中剂量组比较
清燥救肺汤低剂量组ATP 5B mRNA、COXⅣ蛋白表达、ATP含量上调(P<0.01)。结论清燥救肺汤能抑制肺癌细胞增殖
其机制可能与抑制氧化磷酸化能量代谢途径关键限速酶COXⅣ蛋白表达
降低线粒体膜电位
抑制ATP 5B活性
减少ATP生成及升高AMP/ATP值有关。
Objective To explore the underlying mechanism of Qingzao Jiufei Decoction(清燥救肺汤
QZJFD) in the treatment of lung carcinoma. Methods Fifty C57 BL/6 J male mice were randomly divided into five groups
with 10 mice in each group. The groups included a model group
a cyclophosphamide(CTX) group
and QZJFD high
medium and low dose group. Lewis lung carcinoma cells were used to establish the mice tumor-bearing models of lung carcinoma. Twenty-four hours later
the model group was administered with 0.2 ml normal saline by gavage
twice daily; CTX group was injected intraperitoneally with 50 mg/kg cyclophosphamide
once every other day; the high
medium and low dose QZJFD group received 11
5.5 and 2.75 g/(kg·d) Qingzao Jiufei Decoction by gavage respectively
twice daily. QZFJD groups were given intragastric administration 14 days before and after the modelling
while other groups only received treatment 24 hours after the modelling for 14 days. The mRNA expression of adenosine triphosphate synthase β subunit(ATP 5 B) in lung carcinoma tissues was detected by real-time PCR; the protein expression of cytochrome c oxidase subunit Ⅳ(COX Ⅳ) was detected by western blotting; the adenosine monophosphate(AMP)
adenosine diphosphate(ADP)
and adenosine triphosphate(ATP) were detected by high performance liquid chromatography. JC-1 method was used to detect the changes of mitochondrial membrane potential(MMP). Results The expression of ATP 5 B mRNA
COX Ⅳ protein
ATP
and MMP significantly reduced in CTX group and QZJFD high and medium dose group compared to those in model group(P<0.05 or P<0.01). Compared to those in CTX group
the expression of ATP 5 B mRNA and COX IV protein in all QZJFD groups
ATP and MMP levels in QZJFD medium and low dose group
as well as AMP/ATP in QZJFD high dose group significantly increased
while AMP/ATP in QZJFD medium and low dose group
and ADP/ATP in QZJFD low dose group significantly decreased(P<0.05 or P<0.01). Compared to QZJFD high dose group
both the medium and low dose group had higher expression of ATP 5 B mRNA and COX Ⅳ protein
higher ATP level
and lower levels of AMP/ATP and ADP/ATP
and the low dose group also had significantly higher MMP level(P<0.05 or P<0.01). A statistically significant increase was seen in QZJFD low dose group when compared to the medium dose group in terms of ATP 5 B mRNA and COX Ⅳ protein expression
and the ATP level(P<0.01). Conclusion QZJFD may inhibit the proliferation of lung carcinoma cells
by inhibiting the protein expression of COX Ⅳ
the key rate-limiting enzyme of oxidative phosphorylation energy metabolism
reducing mitochondrial membrane potential
inhibiting ATP 5 B activity
reducing ATP production
and increasing the ratio of AMP/ATP.
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