目的 基于過氧化物酶增殖物激活受體γ（PPARγ）通路探討冠心病痰瘀互結證小鼠模型的發(fā)病機制。方法 健康C57BL/6J雄性小鼠隨機分為對照組和假手術組，載脂蛋白E基因敲除（ApoE^-/-）雄性小鼠隨機分為血瘀組、痰濁組、痰瘀互結組。痰濁組和痰瘀互結組小鼠給予高脂飼料喂養(yǎng)12周，其余各組小鼠給予普通飼料喂養(yǎng)12周。在8周末時，對血瘀組和痰瘀互結組小鼠進行冠狀動脈左前降支結扎，假手術組小鼠行穿線不結扎。酶聯(lián)免疫法測定血清白細胞介素-6（IL-6）、內皮素（ET）、血管緊張素II（Ang II）、PPARγ水平，免疫印跡法檢測肝臟組織PPARγ、ATP結合盒轉運蛋白A1（ABCA1）、CD36蛋白表達水平，免疫組化方法檢測主動脈基質金屬蛋白酶-9（MMP-9）、CD40、核轉錄因子-κB（NF-κB）蛋白表達水平。結果 與假手術組比較，各組小鼠血清中IL-6水平未出現(xiàn)顯著變化，痰瘀互結組小鼠血清ET水平顯著升高（P<0.01），血瘀組小鼠血清中Ang II水平顯著升高（P<0.05），痰濁組和痰瘀互結組小鼠血清中Ang II水平顯著升高（P<0.01），PPARγ水平降低；在肝臟組織中，血瘀組、痰濁組和痰瘀互結組小鼠PPARγ、ABCA1蛋白表達水平顯著降低（P<0.01），CD36蛋白表達水平升高；主動脈組織中CD40、MMP-9、NF-κB蛋白表達水平顯著升高（P<0.01）。結論 冠心病痰瘀互結證發(fā)病過程中會造成較為嚴重的動脈粥樣硬化斑塊，可能是通過激活PPARγ通路實現(xiàn)的。

Objective To explore the pathogenesis of phlegm-blood stasis syndrome in mice model of coronary heart disease based on PPAR gamma pathway. Methods Healthy SPF C57BL/6J mice were used in the control group and the sham operation group, and ApoE^-/-mice were used in the blood stasis group, phlegm turbid group and phlegm-blood stasis group. The phlegm turbid group and the phlegm-blood stasis group were fed with high-fat diet for 12 weeks, and the other groups were fed with normal feed for 12 weeks. At the end of the 8th week, the left anterior descending coronary artery was ligated in the blood stasis group and the phlegm-blood stasis group. The sham operation group was not ligated. The levels of IL-6, ET, Ang II and PPARγ in serum were measured by enzyme linked immunosorbent assay, the levels of PPARγ, ABCA1 and CD36 protein in liver tissue were detected by Western blotting, and the levels of CD40, MMP-9 and NF-κB protein in aorta were detected by immunohistochemistry. Results Compared with sham operation group, there was no significant change in serum IL-6, the content of serum ET in the group of phlegm and blood stasis was increased significantly (P<0.01), the content of Ang II in blood stasis group was increased significantly (P<0.05), the content of serum Ang II in phlegm turbid group and phlegm-blood stasis group was increased significantly (P<0.01), and the content of PPARγ was decreased. In liver tissue, the expression levels of PPARγ and ABCA1 protein in blood stasis group, phlegm turbid group and phlegm-blood stasis group were decreased significantly (P<0.01), the expression of CD36 protein was increased. CD40, MMP-9 and NF-κB levels in aorta tissue were increased significantly (P<0.01). Conclusion The phlegm-blood stasis syndrome of coronary heart disease can cause more serious atherosclerotic plaque in the course of its onset. Its mechanism may be through activating PPARγ pathway.

R285.5

國家重點基礎研究計劃項目（2014CB542902）；國家自然科學基金資助項目（81403198）