青蒿素通过HIF-1α/LDHA通路抑制糖酵解改善肺血管重塑

doi:10.3969/j.issn.1006-5725.2025.01.003

摘要/Abstract

摘要：

目的探索青蒿素对野百合碱（MCT）诱导的肺动脉高压（PAH）大鼠血流动力学和血管重塑的作用及分子机制。方法将30只SD大鼠随机分为3组（n = 10）：对照组、MCT诱导的PAH模型组（MCT组，60 mg/kg）、青蒿素干预组（50 mg/kg）。造模28 d后检测大鼠右心室收缩压（RVSP）、肺动脉平均压（mPAP）、心率及右室肥厚指数（RVHI）评估PAH进展，HE染色及α-SMA免疫组化法观察肺血管形态及肌化情况，并计算肺小动脉中膜厚度百分比（WT%）、血管肌层面积百分比（WA%）及肌化血管比例评估肺血管重塑，real time PCR及Western blot分别检测HIF-1α、LDHA的mRNA和蛋白水平，应用丙酮酸及乳酸测试试剂盒检测丙酮酸、乳酸含量。结果与对照组相比，MCT组大鼠RVSP及mPAP升高，心率增快，RVHI增高（P < 0.05）；血管壁明显肌化并增厚，WT%及WA%升高，肺小动脉肌化程度显著增高（P < 0.05），提示PAH模型构建成功。与MCT组相比，青蒿素干预组大鼠RVSP、mPAP、心率及RVHI均降低（P < 0.05），WT%、WA%及肺小动脉肌化程度降低（P < 0.05）。与对照组相比，MCT组大鼠肺组织中HIF-1α、LDHA的mRNA及蛋白水平升高，乳酸和丙酮酸的含量、乳酸/丙酮酸比值升高（P < 0.05）；青蒿素干预组HIF-1α、LDHA mRNA及蛋白水平较MCT组降低，乳酸和丙酮酸的含量、乳酸/丙酮酸比值较MCT组降低（P < 0.05）。结论青蒿素可以改善PAH大鼠的血流动力学及肺血管重塑，其机制可能与抑制HIF-1α/LDHA信号通路介导的糖酵解相关。

关键词: 肺动脉高压, 肺血管重塑, 青蒿素, 糖酵解

Abstract:

Objective Aimed at investigating the effect and molecular mechanism of artemisinin on hemodynamics and vascular remodeling in monocrotaline（MCT） -induced pulmonary arterial hypertension （PAH） rats. Methods 30 male SD rats were randomly divided into 3 groups （n = 10）： control group， MCT-induced PAH group （MCT group， 60 mg/kg） and artemisinin intervention group （50 mg/kg）. At 28 days after modeling， the right ventricular systolic pressure （RVSP）， mean pulmonary artery pressure （mPAP）， heart rate and right ventricular hypertrophy index （RVHI） were measured to evaluate the development of PAH. HE staining and α-SMA immunohistochemistry were used to observe the morphology and assess muscularization of pulmonary arterioles， and the percentage of medial wall thickness （WT%），the percentage of vascular wall area （WA%） and the proportion of muscular vessels were calculated to evaluate the degree of pulmonary vascular remodeling. The mRNA and protein levels of HIF-1α and LDHA were detected by real-time PCR and Western blot， respectively. Pyruvate and lactate concentration in lung tissue was measured using pyruvate and lactateassay kit. Results Compared with the control group， the RVSP， mPAP， heart rate and RVHI were significantly increased in MCT-induced PAH rats （all P < 0.05）. Histological analysis showed that the increasedmedial wall thickness of small pulmonary arteries and vascular muscularization were observed in MCT-treated rats compared with control rats. WT%， WA% and muscularization degrees of pulmonary arterioles were higher in MCT-treated rats than those in the control group （all P < 0.05）， suggesting successful construction of PAH model. Compared with the MCT group， the RVSP， mPAP， heart rate and RVHI decreased in the rats treated with artemisinin（all P < 0.05）， accompanied with lower WT% and WA% （P < 0.05）， and muscularization of pulmonary arterioles was improved （P < 0.05）. Further study showed the mRNA and protein levels ofHIF-1α and LDHA in lung tissue of MCT-induced PAH rats were higher than those in the control group， the content of lactate and pyruvate and the ratio of lactate to pyruvate were higher than that in the control group （all P < 0.05）. However， the mRNA and protein levels of HIF-1α and LDHA in lung tissue of rats treated with artemisinin were lower than those in the MCT group， the content of lactate and pyruvate and the ratio of lactate to pyruvate were lower than that in the MCT group （all P<0.05）. Conclusion Artemisinin improves hemodynamic and pulmonary vascular remodeling in PAH rats through inhibiting HIF-1α/LDHA signaling pathway-mediated glycolysis.

Key words: pulmonary arterial hypertension, pulmonary vascular remodeling, artemisinin, glycolysis

中图分类号:

R543.2

史文花,陈玉倩,张永红,李丛,翟翠,杨妮,柯蕊. 青蒿素通过HIF-1α/LDHA通路抑制糖酵解改善肺血管重塑[J]. 实用医学杂志, 2025, 41(1): 15-22.

Wenhua SHI,Yuqian CHEN,Yonghong ZHANG,Cong LI,Cui ZHAI,Ni YANG,Rui KE. A study of underlying mechanisms of artemisinin inhibiting glycolysis through HIF-1α/LDHA pathway to improve pulmonary vascular remodeling[J]. The Journal of Practical Medicine, 2025, 41(1): 15-22.

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引物名称	核苷酸序列
HIF-1α	上游：5′-ACCTTCATCGGAAACTCCAAAG-3′
HIF-1α	下游：5′-ACTGTTAGGCTCAGGTGAACT-3′
LDHA	上游：5′-GTTGTTGGGGTTGGTGCTGT-3′
LDHA	下游：5′-TCGCCCTTGAGTTTGTCTTC-3′
β-actin	上游：5′-TTACTGCCCTGGCTCCTAG-3′
β-actin	下游：5′-CGTACTCCTGCTTGCTGATC-3′

组别	RVSP/mmHg	mPAP/mmHg	心率 /（次/min）	RVHI /%
对照组	19.56 ± 2.43	17.16 ± 1.96	254 ± 15	0.32 ± 0.05
MCT组	48.51 ± 3.21^*	37.16 ± 2.79^*	383 ± 19^*	0.56 ± 0.04^*
青蒿素干预组	31.02 ± 3.52^#	22.25 ± 1.80^#	278 ± 16^#	0.40 ± 0.03^#
F值	66.67	72.65	54.09	27.16
P值	0.000	0.000	0.000	0.001

组别	MT	WA	肌化程度
对照组	12.96 ± 3.47	16.27 ± 4.03	17.93 ± 3.10
MCT组	35.82 ± 4.89^*	42.15 ± 5.05^*	50.48 ± 4.66^*
青蒿素干预组	20.73 ± 2.96^#	30.06 ± 3.95^#	28.39 ± 4.52^#
F值	23.99	34.73	48.63
P值	0.001	0.001	0.000

组别	HIF-1α mRNA	LDHA mRNA	HIF-1α蛋白	LDHA蛋白
对照组	1.00 ± 0.12	1.00 ± 0.13	1.00 ± 0.17	1.00 ± 0.09
MCT组	1.94 ± 0.14^*	1.73 ± 0.16^*	2.12 ± 0.19^*	1.85 ± 0.10^*
青蒿素干预组	1.25 ± 0.10^#	1.19 ± 0.15^#	1.16 ± 0.20^#	1.31 ± 0.11^#
F值	24.52	18.67	44.19	32.68
P值	0.000	0.001	0.000	0.001

组别	乳酸/（mmol/L）	丙酮酸/（μmol/L）	乳酸/丙酮酸
对照组	0.88 ± 0.16	58.13 ± 6.73	15.18 ± 9.14
MCT组	1.91 ± 0.15^*	71.36 ± 8.92^*	26.86 ± 8.25^*
青蒿素干预组	1.13 ± 0.13^#	61.17 ± 9.24^#	18.55 ± 9.33^#
F值	53.01	25.18	38.42
P值	0.000	0.001	0.000