基于microRNA和Rho激酶信号通路研究滇黄芩有效成分黄芩素抑制低氧性肺动脉高压的多靶点作用机制

Yunnan is located in the plateau and the prevalence rate of hypoxic pulmonary hypertension (HPH) is high. But the price of drugs which is available for the treatment of HPH is expensive and the effect is poor. Therefore, it is necessary to find effective and cheap drug to treat HPH. Our previous study found that microRNA, Rho kinase and NF- kappa B play a key role in the occurrence and development of HPH. Baicalein is one of the main pharmacological component of Scutellaria amoena C.H.Wright of traditional Chinese medicine. Baicalein can regulate the above factors and play the pleiotropic effects of inhibition of proliferation, dilation of blood vessels, anti-inflammatory in many diseases such as cancer. But it is not clear whether it can prevent HPH. Preliminary experiments show that it can inhibit hypoxia-induced proliferation of pulmonary artery smooth muscle cells. Based on our results of a previous study and the pleiotropic effects of baicalein, we will reveal the role of baicalein on hypoxia-induced pulmonary artery smooth muscle cells proliferation, apoptosis and migration, and the effects on microRNA, Rho kinase and NF- kappa B expression through flow cytometry, RT-PCR, Western blot. In addition, we will clarify the target of baicalein by gene transfection technology. Finally, we will elucidate the influence of baicalein on HPH rats and expression of these factors in the lung tissues of rats via the model of HPH rats. The aim of our study is to thoroughly reveal the mechanism of baicalein on HPH by multi-way and multi-target and to provide a cheap and effective drug for HPH prevention and treatment.

云南地处高原低氧肺动脉高压（HPH）患病率高，但当前治疗HPH的药物价格昂贵，且效果不佳，因此需要寻找经济有效的药物。我们前期研究发现microRNA、Rho激酶及NF-κB在HPH发生发展中起关键作用。黄芩素是中药滇黄芩的主要药理成分之一，可通过调节上述因子在肿瘤等疾病中发挥抑制增殖、扩张血管、抗炎等多效性作用。但其是否可防治HPH，目前尚不明确。初步实验表明其可抑制低氧诱导的肺动脉平滑肌细胞增殖。基于前期研究及黄芩素的多效性作用，拟应用流式细胞仪、RT-PCR、免疫印迹等方法揭示其对低氧诱导的肺动脉平滑肌细胞增殖、凋亡及迁移的作用，及对microRNA、Rho激酶及NF-κB表达的影响；并以基因转染等技术，进一步阐明其作用靶点；最后阐明其对HPH大鼠模型的作用，及对上述因子在大鼠肺组织中表达的影响。以期深入全面揭示其对HPH多途径、多靶点的作用机制，为HPH提供一种经济有效的防治药物。

云南地处高原低氧肺动脉高压（HPH）患病率高，但当前治疗HPH的药物价格昂贵，且效果不佳，因此需要寻找经济有效的药物。我们前期研究发现microRNA、Rho激酶及NF-κB在HPH发生发展中起关键作用。黄芩素是中药滇黄芩的主要药理成分之一，可通过调节上述因子在肿瘤等疾病中发挥抑制增殖、扩张血管、抗炎等多效性作用。但其是否可防治HPH，目前尚不明确。初步实验表明其可抑制低氧诱导的肺动脉平滑肌细胞增殖。基于前期研究及黄芩素的多效性作用，拟应用流式细胞仪、RT-PCR、免疫印迹等方法揭示其对低氧诱导的肺动脉平滑肌细胞增殖、凋亡及迁移的作用，及对microRNA、Rho激酶及NF-kb表达的影响。本项目研究发现：（1）低氧可致PASMCs和HPH的大鼠 miR-214水平升高， miR-199 水平降低，提示低氧可致PASMCs miRNA水平的改变；（2）明确了黄芩素可以使低氧对HASMCs miR-199、miR-214产生的变化发生逆转，通过过表达或干扰miR-199、miR-214发现黄芩素可以通过影响miR-199、miR-214的表达来来减轻低氧对PASMCs的影响；（3）体内体外实验证实：黄芩素可以下调miR-214和上调miR-199，并且可以影响Rho激酶和金属蛋白的表达。研究结果将为HPH提出了新的治疗靶点，更新了HPH发病机制理论，也将为HPH的防治提供新的思路。