目的 研究丹红化瘀口服液对气滞血瘀证大鼠眼结膜微循环的改善作用及机制。方法 采用复合因素(夹尾、束缚、冰水浴等)建立气滞血瘀证大鼠模型,造模同时给予丹红化瘀口服液和阿司匹林进行干预,3周后观察大鼠眼结膜血管数、微血管交叉数、微血管管径及血流速度;采用ELISA法检测大鼠血清中6-酮-前列腺素F1α(6-keto-prostaglandin F1α,6-keto-F1α)、血栓素B₂(thromboxane B₂,TXB₂)、血管内皮生长因子(vascular endothelial growth factor,VEGF)、环磷酸腺苷(cyclic adenosine monophosphate,cAMP)、环磷酸鸟苷(cyclic guanosine monophosphate,cGMP)水平及诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)活性;采用微板法测定大鼠血清中一氧化氮(nitric oxide,NO)水平;采用Western blotting法检测大鼠眼结膜组织磷酸化cAMP反应元件结合蛋白(phosphorylated cAMP response-element binding protein,p-CREB)和iNOS蛋白表达情况。结果 与模型组比较,丹红化瘀口服液组大鼠微血管管径显著增大(P<0.05),血流速度加快(P<0.05、0.01),血管数和微血管交叉数显著增多(P<0.05、0.01);血小板最大聚集率及血清中TXB₂/6-keto-F1α、VEGF、cAMP/cGMP、NO水平和iNOS活性均显著降低(P<0.05、0.01);眼结膜组织p-CREB和iNOS蛋白表达水平显著降低(P<0.05、0.01)。结论 丹红化瘀口服液对气滞血瘀模型大鼠的眼结膜微循环有改善作用,其作用机制可能与抑制视网膜组织血栓形成、减少细胞自由基生成、抗炎和抑制血管生成有关。

Objective To study the effect and mechanism of Danhong Huayu Oral Liquid (丹红化瘀口服液) on conjunctival microcirculation in rats with qi stagnation and blood stasis. Methods Rats model with qi stagnation and blood stasis syndrome was established by compound factors (tail clip, restraint, ice water bath, etc.), at the same time, Danhong Huayu Oral Liquid and aspirin were given for intervention. After 3 weeks, number of blood vessels, number of microvascular crossings, microvascular diameter and blood flow rate of conjunctiva were observed; ELISA method was used to detect 6-keto-prostaglandin F1α (6-keto-F1α), thromboxane B₂ (TXB₂), vascular endothelial growth factor (VEGF), cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP) levels and inducible nitric oxide synthase (iNOS) activity in serum of rats; Nitric oxide (NO) level in serum of rats was determined by microplate method; Phosphorylated cAMP response-element binding protein (p-CREB) and iNOS protein expression in eye conjunctiva of rats was detected by Western blotting. Results Compared with model group, number of blood vessels, number of microvascular crossings, microvascular diameter and blood flow rate in Danhong Huayu Oral Liquid group were significantly increased (P < 0.05, 0.01); Maximum platelet aggregation rate and levels of TXB₂/6-keto-F1α, VEGF, cAMP/cGMP, NO and iNOS activity in serum were significantly decreased (P < 0.05, 0.01); p-CREB and iNOS protein expressions in conjunctiva of rats were significantly decreased (P < 0.05, 0.01). Conclusion Danhong Huayu Oral Liquid can improve the conjunctival microcirculation of qi stagnation and blood stasis model rats, and its mechanism may be related to inhibiting retinal tissue thrombosis, reducing cell free radical generation, anti-inflammation and inhibiting angiogenesis.

R285.5

广州市珠江科技新星专项资助（201806010082）；广州市科学技术协会2019年“广州市院士专家工作站”建设项目资助（600180）；中国医学科学院医学与健康科技创新工程项目资助（2019-I2M-5-020）