槌果藤提取物对CIA小鼠模型Treg和Th17细胞及相关细胞因子的影响

Effect of Capparis Spinosa L.Extract on Trag and Th17 Cells and Related Cytokines in CIA Mice

目的探讨槌果藤对胶原诱导关节炎(CIA)模型小鼠全血和脾脏的Treg和Th17细胞比例及IL-17和TGFβ1水平的影响。方法 25只DBA/1J小鼠,采用牛Ⅱ型胶原蛋白制备CIA小鼠模型后根据踝关节病变总积分分为正常组(n=5)、生理盐水组(n=5)、槌果藤低剂量组(0.5g/kg,n=5)、中剂量组(1.0g/kg,n=5)和高剂量组(1.5g/kg,n=5)。免疫后第21天开始给予槌果藤提取物灌胃,共4周。采用细胞流式仪检测小鼠血和脾脏Treg和Th17细胞百分比,ELISA法检测血IL-17和TGFβ1水平。结果 (1)牛Ⅱ型胶原蛋白诱导的CIA小鼠3周后开始首只小鼠足趾出现红肿,第7周到达高峰,小鼠造模成功率达90%以上。(2)与正常组比较,生理盐水组小鼠血和脾脏Treg细胞百分比明显下降[血:(1.17±0.36)%比(7.09±3.86)%,P<0.01);脾脏:(1.56±0.48)%比(8.03±1.75)%,P<0.01)];Th17细胞百分比显著上升[血:(3.96±2.05)%比(0.57±1.60)%,P<0.01);脾脏:(8.32±1.62)%比(0.94±0.30)%,P<0.01)];与生理盐水组比较,槌果藤低、中、高剂量组血、脾脏Treg细胞百分比上升[血:(2.30±0.21)%、(3.78±1.68)%、(4.51±1.58)%比(1.17±0.36)%,P均<0.01);脾脏:(2.75±0.74)%、(4.19±1.45)%、(5.28±1.76)%比(1.56±0.48)%,P均<0.01)],Th17细胞下降[血:(1.78±0.50)%、(1.35±0.36)%、(0.91±0.31)%比(3.96±2.05)%,P均<0.01;脾脏:(3.66±1.69)%、(2.67±0.74)%、(1.70±0.44)%比(8.32±1.62)%,P均<0.01];(3)与正常组比较,生理盐水组小鼠血清IL-17水平明显上升[(31.81±7.33)pg/mL比(12.26±4.89)pg/mL,P<0.01],TGFβ1水平明显下降[(191.62±112.47)pg/mL比(637.33±97.42)pg/mL,P<0.01];与生理盐水组比较,槌果藤低、中、高剂量组血清IL-17水平显著降低[(17.16±2.34)pg/mL、(16.02±2.78)、(10.46±1.83)pg/mL比(31.81±7.33)pg/mL,P<0.01],TGFβ1水平显著升高[(342.63±41.78)pg/mL、(484.76±43.64)pg/mL、(500.83±40.80)pg/mL比(191.62±112.47)pg/mL,P<0.01]。结论槌果藤提取物对CIA模型小鼠血和脾脏Treg和Th17细胞比例及血IL-17和TGFβ1水平具有调节作用。

Objective To investigate the effect of Capparis spinosa L. extract on the percentages of Treg/Th17 cells and the level of IL-17 and TGFβ1 in mice with collagen-induced arthritis（CIA）. Methods CIA was induced in 25 male DBA/1 J mice by immunization with bovine type II collagen. According to mean arthritic index（MAI）the successful model mice were divided into 5 groups： normal group（n=5）, control group（n=5, equivalent physiolog ical saline）, low-, medium-, and high-dose Capparis spinosa L. extract group（n=5 in each group, daily dose of 0.5,1.0, and 1.5g/kg, respectively）. Capparis spinosa L. extract was given by gavage on Day 21 after immunization and lasted 4 weeks. The percentages of Treg and Th17 cells in blood and spleen were detected by flow cytometry, and the serum levels of TGFβ1 and IL-17 were measured by ELISA. Results The first signs of arthritis appeared in mice 3 weeks after immunization and continued to 7 weeks. The incidence of CIA in mice reached more than 90%.Compared with normal group, control group had significantly decreased percentage of Treg cells（blood： 1.17%±0.36%vs 7.09% ±3.86%; spleen： 1.56% ±0.48% vs 8.03% ±1.75%; all P 0.01） and increased percentage of Th17（blood：3.96%±2.05% vs 0.57%±1.60%; spleen： 8.32%±1.62% vs 0.94%±0.30%; all P〈0.01）. The low-, medium- and highdose drug groups had increased percentages of Treg and decreased percentages of Th17 against control group（blood Treg： 2.30%±0.21%, 3.78%±1.68%, 4.51%±1.58% vs 1.17%±0.36%; spleen Treg： 2.75%±0.74%, 4.19%±1.45%, 5.28%±1.76% vs 1.56%±0.48%; blood Th17： 1.78%±0.50%, 1.35%±0.36%, 0.91%±0.31% vs 3.96%±2.05%; spleen Th17：3.66%±1.69%, 2.67%±0.74%, 1.70%±0.44% vs 8.32±1.62%; all P〈0.01）. Compared with normal group, the serum level of IL-17 increased（31.81±7.33pg/mL vs 12.26±4.89pg/mL, P〈0.01） and TGFβ1 decreased（191.62±112.47pg/mL vs 637.33±97.42 pg/mL, P〈0.01） in control group; compared with control group, the serum levels of IL-17 decreased（17.16±2.34pg/mL, 16.02±2.78pg/mL, 10.46±1.83pg/mL vs 31.81±7.33pg/mL; P〈0.01） and the level of TGFβ1 increased（342.63±41.78pg/mL, 484.76±43.64pg/mL, 500.83±40.80pg/mL vs 191.62±112.47pg/mL; P〈0.01） in low-, medium, and high-dose drug groups. Conclusion Capparis spinosa L. Extract can regulate the percentages of Treg and Th17 and the serum levels of IL-17 and TGFβ1 cytokines in mice with CIA.