摘要
目的 探讨芍药苷(PAE)对血栓闭塞性脉管炎(TAO)大鼠的治疗作用及作用机制.方法 通过月桂酸钠注射法建立TAO大鼠模型.将大鼠随机分为假手术组(腹腔注射适量0.9％NaCl)、模型组(腹腔注射适量0.9％NaCl)、低剂量实验组(腹腔内注射5 mg·kg-1·d-1 PAE)、高剂量实验组(腹腔内注射20 mg·kg-1·d-1 PAE)、高剂量+激动药剂(腹腔注射 20 mg·kg-1·d-1PAE+尾静脉注射10 ng·mL-1·kg-1·d-1 740 Y-P)组.用磁珠凝固法检测凝血酶时间(TT);用酶联免疫吸附测定试剂盒检测白细胞介素(IL)-1β、内皮素1(ET-1)水平;用蛋白质印迹法检测磷脂酰肌醇3-激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶 B(AKT)、p-AKT、核因子(NF)-κB p65、p-NF-κB p65蛋白的表达水平.结果 假手术组、模型组、低剂量实验组、高剂量实验组、高剂量+激动药组的 TT 分别为(14.88±1.32)、(10.02±0.95)、(12.65±1.22)、(14.70±1.36)和(10.64±1.21)s,IL-1β 分别为(154.23±13.45)、(356.69±31.17)、(268.62±23.58)、(199.64±20.87)和(337.48±31.46)pg·mL-1,ET-1 分别为(6.78±0.68)、(14.43±1.14)、(11.23±1.07)、(8.20±0.81)和(13.33±1.27)pg·mL1,p-PI3K/PI3K 分别为 0.36±0.04、0.76±0.07、0.59±0.05、0.44±0.04 和 0.69±0.07,p-AKT/AKT 分别为0.52±0.05、0.90±0.09、0.74±0.08、0.61±0.06 和 0.86±0.08,p-NF-κB p65/NF-κB p65 分别为 0.28±0.03、0.95±0.04、0.69±0.07、0.35±0.05 和0.87±0.08.模型组的上述指标与假手术组比较,在统计学上差异均有统计学意义(均P＜0.05);低、高剂量实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P＜0.05);高剂量+激动药组的上述指标与高剂量实验组比较,在统计学上差异均有统计学意义(均P＜0.05).结论 PAE可能通过抑制PI3K/AKT/NF-KB信号通路改善TAO大鼠的疾病进展.
Abstract
Objective To investigate the therapeutic effect and mechanism of paeoniflorin(PAE)on thrombosis angiitis obliterans(TAO)in rats.Methods TAO rat model was established by sodium laurate injection.Rats were randomly divided into sham operation group(intraperitoneal injection of 0.9％NaCl),model group(intraperitoneal injection of 0.9％NaCl),experimental-L,-H groups(intraperitoneal injection of PAE 5,20 mg·kg-1·d-1),experimental-H+agonist group(intraperitoneal injection of 20 mg·kg-1·d-1 PAE+caudal vein injection of 10 ng·mL-1·kg 1·d-1 740 Y-P).Thrombin time(TT)was measured by magnetic bead coagulation;the levels of interleukin(IL)-1 β and endothelin 1(ET-1)were detected by enzyme-linked immunosorbent assay kit;the expression levels of phosphatidylinositol 3-kinase(PI3K),phosphorylated-PI3K(p-PI3 K),protein kinase B(AKT),p-AKT,nuclear factor(NF)-κB p65,p-NF-κB p65 were detected by Western blotting.Results The TT of sham operation group,model group,experimental-L,-H groups and experimental-H+agonist group were(14.88±1.32),(10.02±0.95),(12.65±1.22),(14.70±1.36)and(10.64±1.21)s;IL-1β were(154.23±13.45),(356.69±31.17),(268.62±23.58),(199.64±20.87)and(337.48±31.46)pg·mL-1;ET-1 were(6.78±0.68),(14.43±1.14),(11.23±1.07),(8.20±0.81)and(13.33±1.27)pg·mL-1;p-PI3K/PI3K were 0.36±0.04,0.76±0.07,0.59±0.05,0.44±0.04 and 0.69±0.07;p-AKT/AKT were 0.52±0.05,0.90±0.09,0.74±0.08,0.61±0.06 and 0.86±0.08;p-NF-κB p65/NF-κB p65 were 0.28±0.03,0.95±0.04,0.69±0.07,0.35±0.05 and 0.87±0.08,respectively.There were statistically significant differences between model group and sham operation group(all P＜0.05);the above indexes in experimental-L group and experimental-H group were significantly different from those in medel group(all P＜0.05);the above indexes in experimental-H+agonist group were significantly different from those in experimental-H group(all P＜0.05).Conclusion PAE may improve disease progression in TAO rats by inhibiting the PI3K/AKT/NF-κB signaling pathway.
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