白芍总苷对慢性免疫性血小板减少症的作用机制

The mechanism of total glucosides of paeony on chronic immune thrombocytopenia

关键词:  白芍总苷  免疫性血小板减少症  小鼠  免疫  血小板

Key words:  total glucosides of paeony  immune thrombocytopenia  mice  immunity  platelet

摘要:

[目的] 探讨白芍总苷对慢性免疫性血小板减少症（ITP）的作用机制。[方法] 无特殊病原体动物（SPF）级BALB/C小鼠60只按照随机数字方法分为对照组（10只）、模型组（9只）、白芍总苷（TGP）低剂量组（10只）、TGP中剂量组（9只）、TGP高剂量组（10只）及西药组（10只），除对照组外均建立ITP动物模型，建模后24 h TGP低、中及高剂量组均灌胃100 mg/kg、200 g/kg及400 mg/kg的TGP溶液，西药组灌胃5 mg/kg的醋酸泼尼松，其余组灌胃等体积的生理盐水，采用全自动生化分析仪检测建模前、建模后24 h、干预1周、干预2周、干预3周、干预4周时血小板计数；流式细胞仪检测辅助性T 细胞17（Th17）、全称调节性T细胞（Teg）百分比；酶联免疫吸附试剂盒检测白细胞介素（IL）-10、IL-17及转化生长因子（TGF）-β1水平；苏木精-伊红（HE）染色观察肝组织及脾组织病理形态并计算脏器指数。[结果] 建模24 h及干预1周，与对照组相比，其余各组小鼠血小板计数均降低（P＜0.05），但组间比较差异无统计学意义（P＞0.05）。干预2周、3周及4周时，与对照组相比，模型组血小板计数降低（P＜0.05），与模型组相比，TGP低、中、高剂量组及西药组血小板计数升高（P＜0.05），TGP低、中、高剂量组及西药组组间比较血小板计数逐渐升高，存在差异（P＜0.05）。与对照组相比，模型组小鼠外周血Treg百分比降低，Th17百分比、肝指数及脾指数升高（P＜0.05）。与模型组相比，TGP低、中、高剂量组及西药组Treg百分比升高，Th17百分比、肝指数及脾指数降低（P＜0.05）。与对照组相比，模型组小鼠外周血IL-10、TGF-β1降低，IL-17升高（P＜0.05）。与模型组相比，TGP低、中、高剂量组及西药组IL-10、TGF-β1升高，IL-17降低（P＜0.05）。IL-17在TGP低、中、高剂量组组间比较差异有统计学意义（P＜0.05），TGF-β1在TGP低、中、高剂量组、西药组组间比较无差异（P＞0.05），IL-17在TGP高剂量组与西药组比较无差异（P＞0.05）。[结论] 白芍总苷可改善慢性ITP小鼠免疫功能，提高血小板计数，减少肝脾损伤。

Abstract:

[Objective] To investigate the mechanism of total glucosides of paeony on chronic immune thrombocytopenia. [Methods] Sixty BALB/C mice with SPF grade were divided into control group（n=10），model group（n=9），total glucosides of peony（TGP） low dose group（n=10），TGP medium dose group（n=9），TGP high dose group，and Western medicine group（n=10） according to random number method. Immune thrombocytopenia（ITP） animal models were established in both the Western medicine group（n=10） and the Western medicine group（n=10） except the control group. After modeling，24 h TGP low，medium，and high dose groups were given a TGP solution of 100 mg/kg，200 g/kg，and 400 mg/kg，while the Western medicine group was given 5 mg/kg prednisone acetate. The other groups were given the same volume of normal saline，and the platelet count was detected by automatic biochemical analyzer before modeling，24 h after modeling，1 week of intervention，2 weeks of intervention，3 weeks of intervention，and 4 weeks of intervention. The percentages of helper T cells 17（Th17） and regulatory T cells（Teg） were detected by flow cytometry. The levels of interleukin-10，IL-17，and transforming growth factor（TGF） -β1 were detected by an enzyme-linked immunosorbent kit. Hematoxylin-eosin（HE） staining was used to observe the pathological morphology of liver and spleen tissue and to calculate the organ index. [Results] At 24 hours after modeling and 1 week of intervention，compared to the control group，the platelet counts in all other groups of mice decreased（P<0.05），but there was no significant difference among the groups（P>0.05）. At 2，3，and 4 weeks of intervention，compared to the control group，the platelet counts in the model group decreased（P<0.05），while the platelet counts in the TGP low，medium，high dose groups，and Western medicine group increased compared to the model group（P<0.05）. The platelet counts gradually increased in the TGP low，medium，and high dose groups，and Western medicine group，with significant differences observed among the groups（P<0.05）. Compared to the control group，the percentage of Treg in the peripheral blood of mice in the model group decreased，while the percentage of Th17，liver index，and spleen index increased（P<0.05）. Compared to the model group，the TGP low，medium，and high dose groups，and Western medicine group showed an increase in the percentage of Treg，and a decrease in the percentage of Th17，liver index，and spleen index（P<0.05）. Compared to the control group，the levels of IL-10 and TGF-β1 in the peripheral blood of mice in the model group decreased，while IL-17 increased（P<0.05）. Compared to the model group，the TGP low，medium，and high dose groups，and Western medicine group showed an increase in IL-10 and TGF-β1 levels，and a decrease in IL-17 levels（P<0.05）. IL-17 showed differences among the TGP low，medium，and high dose groups（P<0.05），while there was no difference in TGF-β1 levels among the TGP low，medium，high dose groups and western medicine group（P>0.05）. There was no difference in IL-17 levels between the TGP high dose group and the Western medicine group（P>0.05）. [Conclusion] Total glucoside of paeony can improve immune function，increase platelet count，and reduce liver and spleen injury in mice with chronic immune thrombocytopenia.