Abstract：
BACKGROUND: With most of the anti-rheumatic drugs having severe adverse drug reactions and poor tolerance, the active components from natural herbs provides a repository for novel, safe, and effective drug development. Sanguisorba officinalis L. exhibits definite anti-inflammatory capacity, however, whether it has anti-rheumatic effects has not been revealed.
OBJECTIVE: In the present study, the effect of Ziyuglycoside I (Ziyu I), one of the most important active components in Sanguisorba officinalis L., was investigated in treating collagen-induced arthritis (CIA), illuminating its potential pharmacological mechanisms.
METHODS: CIA mice were treated with 5, 10, or 20 mg/kg of Ziyu I or 2 mg/kg of MTX, and clinical manifestations as well as pathological changes were observed. T and B cell viability was determined using cell counting kit-8, plasma autoantibodies and cytokines were tested with ELISA, T and B cell subsets were identified by flow cytometry, Blimp1 expression was detected by RT-qPCR and in situ immunofluorescence. The expression of activation-induced cytidine deaminase (AID) was detected by immunohistochemistry. ERK activation in B cells was verified through western blotting and immunofluorescence. Meanwhile, bioinformatics retrieval and molecular docking/molecular dynamics were used to predict the relationship between Blimp1, ERK and Ziyu I with the pharmacokinetics and toxicity of Ziyu I being evaluated in the ADMETlab Web platform.
RESULTS: Ziyu I treatment effectively alleviated the joint inflammatory manifestation including arthritis index, global scores, swollen joint count and body weight of CIA mice. It improved the pathological changes of joint and spleen of arthritic mice, especially in germinal center formation. Ziyu I displayed a moderate regulatory effect on T cell activation, the percentage of total T and helper T cells, and tumor necrosis factor-α, but transforming growth factor-β was not restored. Increased spleen index, B cell viability and plasma auto-antibody production in CIA mice were significantly reduced by Ziyu I therapy. Of note, we found that Ziyu I administration substantially inhibited the excessive expansion of plasma cells in spleen through preventing the expression of B lymphocyte induced maturation protein 1 (Blimp1) and AID in B cells. Ziyu I was predicted in silico to directly interact with ERK2, and reduce ERK2 activation, contributing to the depressed expression of Blimp1. Moreover, Ziyu I was predicted to have a favorable pharmacokinetic profile and low toxicity.
CONCLUSIONS: Ziyu I effectively ameliorates CIA in mice by inhibiting plasma cell generation through prevention of ERK2-mediated Blimp1 expression in B cells. Therefore, Ziyu I is a promising candidate for anti-arthritic drug development.
OBJECTIVE: In the present study, the effect of Ziyuglycoside I (Ziyu I), one of the most important active components in Sanguisorba officinalis L., was investigated in treating collagen-induced arthritis (CIA), illuminating its potential pharmacological mechanisms.
METHODS: CIA mice were treated with 5, 10, or 20 mg/kg of Ziyu I or 2 mg/kg of MTX, and clinical manifestations as well as pathological changes were observed. T and B cell viability was determined using cell counting kit-8, plasma autoantibodies and cytokines were tested with ELISA, T and B cell subsets were identified by flow cytometry, Blimp1 expression was detected by RT-qPCR and in situ immunofluorescence. The expression of activation-induced cytidine deaminase (AID) was detected by immunohistochemistry. ERK activation in B cells was verified through western blotting and immunofluorescence. Meanwhile, bioinformatics retrieval and molecular docking/molecular dynamics were used to predict the relationship between Blimp1, ERK and Ziyu I with the pharmacokinetics and toxicity of Ziyu I being evaluated in the ADMETlab Web platform.
RESULTS: Ziyu I treatment effectively alleviated the joint inflammatory manifestation including arthritis index, global scores, swollen joint count and body weight of CIA mice. It improved the pathological changes of joint and spleen of arthritic mice, especially in germinal center formation. Ziyu I displayed a moderate regulatory effect on T cell activation, the percentage of total T and helper T cells, and tumor necrosis factor-α, but transforming growth factor-β was not restored. Increased spleen index, B cell viability and plasma auto-antibody production in CIA mice were significantly reduced by Ziyu I therapy. Of note, we found that Ziyu I administration substantially inhibited the excessive expansion of plasma cells in spleen through preventing the expression of B lymphocyte induced maturation protein 1 (Blimp1) and AID in B cells. Ziyu I was predicted in silico to directly interact with ERK2, and reduce ERK2 activation, contributing to the depressed expression of Blimp1. Moreover, Ziyu I was predicted to have a favorable pharmacokinetic profile and low toxicity.
CONCLUSIONS: Ziyu I effectively ameliorates CIA in mice by inhibiting plasma cell generation through prevention of ERK2-mediated Blimp1 expression in B cells. Therefore, Ziyu I is a promising candidate for anti-arthritic drug development.
摘要：
背景：由于大多数抗风湿药具有严重的药物不良反应和较差的耐受性，来自天然草药的活性成分为新颖的，安全,和有效的药物开发。地志表现出一定的抗炎能力，然而,它是否具有抗风湿作用尚未透露。
目的：在本研究中，紫玉苷I（紫玉I）的作用，是地黄中最重要的活性成分之一，在治疗胶原诱导性关节炎（CIA）方面进行了研究，阐明其潜在的药理机制。
方法：CIA小鼠用5、10或20mg/kg的ZiyuI或2mg/kg的MTX治疗，观察临床表现和病理改变。使用细胞计数试剂盒-8测定T细胞和B细胞活力，用ELISA检测血浆自身抗体和细胞因子，通过流式细胞术鉴定T和B细胞亚群，通过RT-qPCR和原位免疫荧光检测Blimp1的表达。免疫组化法检测活化诱导胞苷脱氨酶(AID)的表达。通过蛋白质印迹和免疫荧光验证B细胞中的ERK活化。同时,通过生物信息学检索和分子对接/分子动力学方法预测Blimp1，ERK和ZiyuI之间的关系，并在ADMETlabWeb平台上评估了ZiyuI的药代动力学和毒性。
结果：紫雨I号治疗可有效缓解关节炎症表现，包括关节炎指数，全球得分，CIA小鼠的肿胀关节计数和体重。改善关节炎小鼠关节和脾脏的病理变化，尤其是生发中心的形成。紫玉I对T细胞活化表现出适度的调节作用，总T细胞和辅助性T细胞的百分比，和肿瘤坏死因子-α,但转化生长因子-β未恢复。脾脏指数增加，ZiyuI疗法显着降低了CIA小鼠的B细胞活力和血浆自身抗体的产生。值得注意的是，我们发现，紫玉I号给药通过阻止B细胞中B淋巴细胞诱导成熟蛋白1（Blimp1）和AID的表达而实质上抑制了脾脏中浆细胞的过度扩增。子玉I被预测为直接与ERK2相互作用，并减少ERK2的激活，导致Blimp1的表达降低。此外，据预测，ZiyuI具有良好的药代动力学特征和低毒性。
结论：紫雨I通过阻止ERK2介导的Blimp1在B细胞中的表达，抑制浆细胞的生成，从而有效改善小鼠的CIA。因此,ZiyuI是抗关节炎药物开发的有希望的候选人。
目的：在本研究中，紫玉苷I（紫玉I）的作用，是地黄中最重要的活性成分之一，在治疗胶原诱导性关节炎（CIA）方面进行了研究，阐明其潜在的药理机制。
方法：CIA小鼠用5、10或20mg/kg的ZiyuI或2mg/kg的MTX治疗，观察临床表现和病理改变。使用细胞计数试剂盒-8测定T细胞和B细胞活力，用ELISA检测血浆自身抗体和细胞因子，通过流式细胞术鉴定T和B细胞亚群，通过RT-qPCR和原位免疫荧光检测Blimp1的表达。免疫组化法检测活化诱导胞苷脱氨酶(AID)的表达。通过蛋白质印迹和免疫荧光验证B细胞中的ERK活化。同时,通过生物信息学检索和分子对接/分子动力学方法预测Blimp1，ERK和ZiyuI之间的关系，并在ADMETlabWeb平台上评估了ZiyuI的药代动力学和毒性。
结果：紫雨I号治疗可有效缓解关节炎症表现，包括关节炎指数，全球得分，CIA小鼠的肿胀关节计数和体重。改善关节炎小鼠关节和脾脏的病理变化，尤其是生发中心的形成。紫玉I对T细胞活化表现出适度的调节作用，总T细胞和辅助性T细胞的百分比，和肿瘤坏死因子-α,但转化生长因子-β未恢复。脾脏指数增加，ZiyuI疗法显着降低了CIA小鼠的B细胞活力和血浆自身抗体的产生。值得注意的是，我们发现，紫玉I号给药通过阻止B细胞中B淋巴细胞诱导成熟蛋白1（Blimp1）和AID的表达而实质上抑制了脾脏中浆细胞的过度扩增。子玉I被预测为直接与ERK2相互作用，并减少ERK2的激活，导致Blimp1的表达降低。此外，据预测，ZiyuI具有良好的药代动力学特征和低毒性。
结论：紫雨I通过阻止ERK2介导的Blimp1在B细胞中的表达，抑制浆细胞的生成，从而有效改善小鼠的CIA。因此,ZiyuI是抗关节炎药物开发的有希望的候选人。
