绿原酸对深静脉血栓形成大鼠T细胞亚群失衡的影响

目的：探讨Th1/Th2亚群失衡在大鼠深静脉血栓形成（DVT）中的作用及绿原酸的作用机制。方法：40只DVT大鼠随机分为DVT模型组、绿原酸小剂量组、绿原酸中剂量组、绿原酸大剂量组，每组各10只，分离大鼠脾脏单核细胞，流式细胞术检测各组大鼠Th1亚群（CD4＋TNF-α＋）、Th2亚群（CD4＋IL-4＋）比例，ELISA法检测大鼠外周血IL-2、TNF-α、IL-4、IL-10水平，并分析Th1/Th2比例与血清炎症标记物相关性。结果：与正常组及假手术组比较，DVT模型组大鼠脾脏Th1亚群比例升高（22.71±0.87，%），Th2亚群比例降低（3.06±0.15，%），Th1/Th2升高（7.20±0.45），IL-2、TNF-α表达升高（72.10±7.48，173.50±17.02, pg/mL），IL-4、IL-10表达降低（347.46±11.24、28.55±1.78,pg/mL）；与模型组比较，绿原酸大剂量组大鼠Th1亚群比例降低（14.38±0.41,%），Th2亚群比例升高（4.91±0.14,%），Th1/Th2降低（3.02±0.26），IL-2、TNF-α表达降低（50.22±4.06、110.22±10.32, pg/mL），IL-4、IL-10表达升高（418.72±15.32、9.32±2.83,pg/mL），绿原酸大剂量组疗效优于中、小剂量组（P＜0.05）；Th1/Th2比例与IL-2、TNF-α呈正相关，与IL-4、IL-10呈负相关。结论：大鼠DVT发病过程中存在Th1/Th2亚群失衡，绿原酸诱导T细胞向Th2亚群偏移，逆转Th1/Th2亚群失衡状态，进而减轻炎症反应，抑制血栓形成。

Regulating Effect of Chlorogenic Acid on T cell Subsets Imbalance in Deep Venous Thrombosis of Rats

Objective To evaluate the role of Th1/Th2 shift in deep venous thrombosis （DVT） and elucidate the mechanism of chlorogenic acid in treating DVT in rat models. Method Forty DVT rats were divided randomly into DVT group and chlorogenic acid high dose, middle dose and low dose groups. Additionally, 10 rats with sham operation were taken as sham operation group and 10 normal rats were taken as normal group respectively. Rats of chlorogenic acid group were treated with chlorogenic acid for 7 days, while rats of other groups were treated with physiological saline for the same duration. Then, the proportions of Th1 （CD4＋TNF-α＋） subset and Th2 （CD4＋IL-4＋） subset were detected by flowcytometry. Moreover, the protein expressions of IL-2, TNF-α, IL-4 and IL-10 in rat serum were analyzed by ELISA assay, and the proportion of Th1/ Th2 correlated with serum levels of inflammatory markers was analyzed. Results Compared with normal and sham operation groups, the proportion of Th1 subset was increased while that of Th2 subset was decreased significantly in DVT group（22.71 ± 0.87%,3.06 ± 0.15%）. In addition, protein expression of IL-2 and TNF-α was enhanced （72.10 ± 7.48 173.50 ± 17.02, pg/mL）while that of IL-4 and IL-10 was inhibited markedly in DVT rats （347.46 ± 11.24 28.55 ± 1.78,pg/mL）. After chlorogenic acid administration, the proportion of Th1 subset was down-regulated while that of Th2 subset was up-regulated significantly in DVT rats（14.38 ± 0.4 14.91 ± 0.14, %）. Moreover, the protein expression of＆amp;nbsp;IL-2 and TNF-α decreased while that of IL-4 and IL-10 increased obviously after chlorogenic acid administration, Th1/ Th2 proportional positively correlated with IL-2 and TNF-α, and negatively correlated with IL-4, IL-10（50.22 ± 4.06 110.22 ± 10.32, 418.72 ± 15.32 9.32 ± 2.83,pg/mL）. Conclusion Th1/Th2 shift is involved in DVT. Chlorogenic acid can down-regulate the Th1 subsets and up-regulate the Th2 subsets and exert the curative effect on DVT mice.