目的 通过野百合碱诱导的肺动脉高压(PH)大鼠模型,探讨香青兰总黄酮(DMTF)对PH大鼠的保护作用及作用机制.方法 建立野百合碱诱导的PH大鼠模型,采用超声法检测大鼠心脏功能,观察肺的病理变化并评估PH的发展状态以及肺血管重构程度.使用ELISA法检测血清相关指标,评价DMTF抗炎、抗氧化应激能力;使用Western blot法检测BMPR2、TGF-β1、Smad1/5/8、p-Smad1/5/8、Smad2/3 和p-Smad2/3 的蛋白表达.通过低氧诱导肺动脉平滑肌细胞(PASMCs)增殖模型,并用CCK-8法检测细胞增殖能力;使用流式细胞术检测细胞周期及凋亡,并采用 Western blot 法检测 BMPR2、TGF-β1、Smad1/5/8、p-Smad1/5/8、Smad2/3和p-Smad2/3的蛋白表达.结果 心脏超声和病理检查结果显示,PH大鼠平均肺动脉压力(mPAP)显著升高,血管壁增厚,α-SMA表达增加,血清中ET-1、IL-1β、IL-6、TNF-α和MDA含量升高,SOD、GSH-Px和NO含量降低(P＜0.05).DMTF药物干预后降低mPAP,并减轻或延缓肺动脉血管重构;升高血清SOD、GSH-Px和NO含量,降低ET-1、IL-1β、IL-6、TNF-α 和 MDA 含量;下调 PH 大鼠肺组织中 TGF-β1、p-Smad2/3 蛋白的表达,并上调BMPR2、p-Smad1/5/8蛋白的表达(P＜0.05).CCK-8和流式细胞术结果显示DMTF抑制了低氧诱导的PASMCs过度增殖,并下调TGF-β1、p-Smad2/3蛋白的表达,上调BMPR2、p-Smad1/5/8蛋白的表达(P＜0.05).结论 DMTF可降低肺动脉压,改善肺血管重构,对PH大鼠具有一定的保护作用,其机制可能与抑制炎症反应、减轻氧化应激反应和调节TGF-β/BMP信号通路有关.
Protective effect of total flavones in Dracocephalum Moldavica L.on pulmonary hypertension in rats via modulation of TGF-β/BMP signaling pathway
Objective To establish the rat model of pulmonary hypertension(PH)induced by monocrotaline and to determine the protective effect and mechanism of action of total flavone in Dracocephalum Moldavica L.(DMTF)on PH rats.Methods After the establishment of a monocrotaline-induced PH rat model,the cardiac function of rats was determined by ultrasound.The pathological changes in the lung were observed to evaluate the development of PH and the degree of pulmonary vascular remodeling.The serum nitric oxide(NO),endothelin-1(ET-1),interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione peroxidase(GSH-Px)were determined by ELISA,and bone morphogenetic protein receptor 2(BMPR2),transforming growth factor-β1(TGF-β1),Smad2/3 and p-Smad2/3 were determined by Western blot.The proliferation model of pulmonary artery smooth muscle cells(PASMCs)was induced by hypoxia in vitro,and the cell proliferation ability was determined by CCK-8 method.The cell cycle and apoptosis were detected with flow cytometry,and the protein expression levels of BMPR2,TGF-β1,Smad1/5/8,p-Smad1/5/8,Smad2/3,and p-Smad2/3 were determined by Western blot.Results Echocardiography and pathology showed significantly increased mean pulmonary artery pressure(mPAP)of the PH rats,thickened vascular walls,increased α-SMA expression,elevated serum content of ET-1,IL-1β,IL-6,TNF-α and MDA,and lowered the content of SOD,GSH-Px and NO(P＜0.05).DMTF intervention reduced mPAP,alleviated/delayed pulmonary artery vascular remodeling,elevated the serum content of SOD,GSH-Px and NO,and lowered the content of ET-1,IL-1β,IL-6,TNF-α and MDA,down-regulated the protein expression of TGF-β1 and p-Smad2/3 in the lung tissue of PH rats,and upregulated the protein expression of BMPR2(P＜0.05).CCK-8 and flow cytometry showed that DMTF inhibited hypoxia-induced hyperproliferation of PASMCs,reduced the protein expression of TGF-β1,p-Smad2/3,and promoted the protein expression of BMPR2 and p-Smad1/5/8(P＜0.05).Conclusion DMTF can reduce the pulmonary artery pressure and improve pulmonary vascular remodeling.The mechanism of protective effect on PH rats may be related to inhibiting inflammation,alleviating oxidative stress reactions and regulating the TGF-β/BMP signaling pathway.
total flavone in Dracocephalum Moldavica L.pulmonary hypertensionBMPR2TGF-β1
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