葛根芩连汤含药血清调节缺氧诱导L02细胞糖代谢的影响及代谢机制

doi:10.3969/j.issn.1006-5725.2025.07.002

Abstract

Abstract:

Objective The study aimedto investigate the effects and metabolic mechanisms of Gegen Qinlian Decoction （GQD） containing serum on hypoxia?induced glucose metabolism in L02 cells. Methods The effects of five hypoxia durations （6， 12， 18， 24， and 48 hours） on glucose consumption and cell viability of L02 cells were examined under hypoxic conditions to determine the optimal hypoxia time. Normal hepatocytes served as the normal control group. L02 cells with hypoxia?induced reduction in glucose consumption were divided into several groups： hypoxia group， metformin 2 mmol/L group， and 25 g/kg GQD groups treated with 5%， 10%， and 15% GQD containing serum. Glucose consumption was used as an indicator of drug efficacy. High?resolution liquid chromatography tandem quadrupole time?of?flight mass spectrometry （UPLC?Q?TOF?MS） was employed to collect metabolite signals from each group. Data were analyzed by using Progenesis QI software， and potential biomarkers were identified through online databases such as HMDB. Finally， metabolic pathways of potential biomarkers were analyzed via the Metabo Analyst 5.0 website. Results An 18 hour hypoxia period was identified as the optimal duration for the replication of the hypoxia?induced L02 cell model. GQD containing serums at 5% and 10% significantly increased glucose consumption in hypoxia?induced L02 cells after 18 hours. 14 biomarkers of hypoxia?induced L02 cells were identified， with the levels of 13 biomarkers significantly increased and 1 biomarker significantly decreased. GQD containing serum notably regulated the levels of 3 biomarkers. Conclusion GQD containing serum might improve hypoxia?induced abnormal glucose metabolism in L02 cells and enhance glucose consumption by modulating glycerophospholipid metabolism， glycosylphosphatidylinositol biosynthesis， and sphingolipid metabolism.

Key words: Gegen Qinlian Decoction containing serum, L02 cells, hypoxia, glucose metabolism, metabolomics

CLC Number:

R285.5

Yan YOU,Hongjing CUI,Chengcheng PENG,Li JIANG,Qiyun ZHANG,Bingtao LI,Guoliang XU. The analysis of effect of serum containing Gegen Qinlian Decoction on regulating hypoxia-induced glucose metabolism in L02 Cells and related metabolic mechanisms[J]. The Journal of Practical Medicine, 2025, 41(7): 936-943.

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组别	组成
正常对照组	15%空白血清+10%FBS培养基（常氧）
缺氧组	15%空白血清+10%FBS培养基（1%氧浓度）
二甲双胍组	15%空白血清+10%FBS培养基+2 mmol/L二甲双胍溶液（1%氧浓度）
GQD低剂量组（GQD-L）	10%空白血清+5%含药血清+10%FBS培养基（1%氧浓度）
GQD中剂量组（GQD-M）	5%空白血清+10%含药血清+10%FBS培养基（1%氧浓度）
GQD高剂量组（GQD-H）	15%含药血清+10%FBS培养基（1%氧浓度）

质谱条件	正离子模式	负离子模式
离子化方式	Dual ESI源	Dual ESI源
电离源温度	100 ℃	100 ℃
锥孔气流速	50 L/h	50 L/h
去溶剂温度	400 ℃	400 ℃
毛细管电压	3.5 kV	2.5 kV
锥孔电压	40 V	40 V
扫描范围	50 ~ 1 000 Da	50 ~ 1 000 Da

模式	关键生物标记物	HMDB号	m/z	t_R/min	变化趋势
模式	关键生物标记物	HMDB号	m/z	t_R/min	缺氧组 vs. 正常对照组
正离子	戊酸糠酯	HMDB37727	114.0364	11.56	↑^?
	CE（22：2（13Z，16Z））	HMDB06737	722.6908	0.45	↑^?
	PC（14：0/24：0）	HMDB07893	782.6435	0.47	↑^?
	SM（d18：1/24：1（15Z））	HMDB12107	851.6434	0.47	↑^?
	琥珀酰丙酮	HMDB00635	176.0909	0.4	↑^?
	SM（d18：1/22：1（13Z））	HMDB12104	784.6661	0.45	↑^?
	CMP	HMDB01202	615.1244	6.16	↑^?
	PGP（a-13：0/18：2（9Z，11Z））	HMDB0116513	863.4516	9.68	↑^?
负离子	PS（14：0/18：0）	HMDB12334	780.5069	0.45	↑^?
	PC（14：0/20：4（5Z，8Z，11Z，14Z））	HMDB07883	774.5044	0.45	↑^?
	脱氧胞苷	HMDB00014	306.0092	0.73	↓^?
	PE（15：0/20：0）	HMDB08899	714.5415	0.47	↑^?
	GlcCer（d18：1/18：0）	HMDB04972	748.5686	0.45	↑^?
	PE（16：1（9Z）/24：1（15Z））	HMDB08981	878.5373	0.45	↑^?

模式	代谢物名称	峰面积
模式	代谢物名称	正常对照组	缺氧组	GQD-L组	GQD-M组	GQD-H组
负离子	PE（15：0/20：0）	231.19 ± 55.13	564.38 ± 103.14^?	405.97 ± 25.51^#	748.92 ± 255.7	505.65 ± 204.58
	GlcCer（d18：1/18：0）	670.56 ± 312.64	1 412.56 ± 259.35^?	1 003.14 ± 49.54^#	2 050.09 ± 790.94	1 382.13 ± 516.55
	PE（16：1（9Z）/24：1（15Z））	835.12 ± 188.89	1 426.25 ± 244.79^?	1 008.15 ± 48.65^#	1 857.58 ± 618.74	1 343.45 ± 510.98

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The analysis of effect of serum containing Gegen Qinlian Decoction on regulating hypoxia-induced glucose metabolism in L02 Cells and related metabolic mechanisms

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扫描模式	时间（t_R）/min	A相/%	B相/%
正离子模式	0	99	1
	2	99	1
	15	64	36
	18	40	60
	20	20	70
	22	20	80
	24	99	1
负离子模式	0	99	1
	2	99	1
	15	64	36
	18	40	60
	20	20	70
	22	20	80
	24	99	1