中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (36): 7735-7742.doi: 10.12307/2025.558

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

骨髓增生异常综合征模型大鼠骨髓造血：活髓方干预免疫检查点的作用机制

卓秋燕1，蒋  群1，夏  思1，卢诗颖1，刘燕娣2，戴 媺1   

  1. 广州医科大学附属中医医院，1血液科，2病理科，广东省广州市   510130
  • 收稿日期:2024-07-20 接受日期:2024-09-21 出版日期:2025-12-28 发布日期:2025-03-05
  • 通讯作者: 戴媺，硕士，主任中医师，广州医科大学附属中医医院血液科，广东省广州市 510130
  • 作者简介:卓秋燕，女，1992年生，广东省湛江市人，汉族，2019年广州中医药大学毕业，硕士，医师，主要从事中医药治疗血液疾病研究。
  • 基金资助:
    广州市科技厅项目(202102080138)，项目负责人：戴媺；广州市三级名中医工作室建设项目(穗卫中医[2022]3号)，项目负责人：戴媺

Bone marrow hematopoiesis in rats with myelodysplastic syndrome: action mechanism of Huosui Formula in intervening immune checkpoints

Zhuo Qiuyan1, Jiang Qun1, Xia Si1, Lu Shiying1, Liu Yandi2, Dai Mei1   

  1. 1Department of Hematology, 2Department of Pathology, Affiliated Traditional Chinese Medicine Hospital, Guangzhou Medical University, Guangzhou 510130, Guangdong Province, China
  • Received:2024-07-20 Accepted:2024-09-21 Online:2025-12-28 Published:2025-03-05
  • Contact: Dai Mei, MS, Chief physician, Department of Hematology, Affiliated Traditional Chinese Medicine Hospital, Guangzhou Medical University, Guangzhou 510130, Guangdong Province, China
  • About author:Zhuo Qiuyan, MS, Physician, Department of Hematology, Affiliated Traditional Chinese Medicine Hospital, Guangzhou Medical University, Guangzhou 510130, Guangdong Province, China
  • Supported by:
    Guangzhou Science and Technology Department Project, No. 202102080138 (to DM); Guangzhou Municipal Tier-3 Famous Traditional Chinese Medicine Studio Construction Project, No. [2022]3 (to DM)

PDF

131

摘要：

文题释义：

免疫检查点：逆转肿瘤免疫抑制功能的CTLA-4、PD-1/PD-L1等免疫检查点在血液肿瘤及实体肿瘤治疗中取得了令人振奋的效果，成为治疗癌症的新手段，并在骨髓增生异常综合征中显示出一定的优势。
活髓方：名中医陈志雄教授临床应用补虚解毒法的经验方联合西药治疗骨髓增生异常综合征，对免疫及造血调节起到增效减毒的协同作用，高效低毒的经验方剂活髓方能改善骨髓增生异常综合征患者的生存预后。

摘要
背景：既往研究表明经验方剂活髓方对骨髓增生异常综合征患者的免疫及造血调节起到增效减毒的协同作用，但具体机制尚未明确。
目的：探讨活髓方对骨髓增生异常综合征模型大鼠骨髓造血的影响及机制。
方法：70只SD大鼠采用随机数字表法分为正常对照组10只、模型组15只、西药组15只、活髓方低剂量组15只、活髓方高剂量组15只。除正常对照组外，其余4组采用尾静脉注射二甲基苯蒽诱导建立大鼠骨髓增生异常综合征模型，造模后正常对照组、模型组给予生理盐水，西药组给予沙利度胺胶囊10 mg/kg、维A酸片4 mg/kg，活髓方低、高剂量组分别给予活髓方1.5，6 g/kg，每天灌胃1次，连续给药28 d，取外周血和股骨骨髓组织，检测外周血常规、骨髓活检造血增殖情况，采用流式细胞术检测T淋巴细胞亚群及T淋巴细胞上CTLA-4和PD-1的表达。 
结果与结论：①与正常对照组相比，模型组外周血白细胞、中性粒细胞、血红蛋白、血小板和CD4+、CD4+/CD8+水平显著下降(P < 0.05)，CD4+PD-1+、CD8+PD-1+、CD4+CTLA-4+和CD8+CTLA-4+表达显著上调(P < 0.05)；②与模型组相比，各给药组的骨髓增生程度均有改善，但组间无统计学差异(P > 0.05)；③与模型组相比，活髓方高剂量组白细胞、血红蛋白、血小板和CD4+、CD4+/CD8+水平显著升高(P < 0.05)，CD8+水平显著降低(P < 0.05)，CD4+PD-1+、CD8+PD-1+、CD4+CTLA-4+和CD8+CTLA-4+表达水平下降但无统计意义(P > 0.05)；④西药组与活髓方高剂量组显示出相似的疗效；活髓方高剂量组较低剂量组各指标改善均有优势。结果表明，活髓方可改善骨髓增生异常综合征模型大鼠的骨髓造血，提高CD4+、CD4+/CD8+水平，下调CD4+PD-1+、CD8+PD-1+、CD4+CTLA-4+和CD8+CTLA-4+表达，可能与免疫负调控机制相关。

关键词: 骨髓增生异常综合征, 活髓方, 骨髓造血, 免疫检查点, 外周血T淋巴细胞亚群, CTLA-4, PD-1, SD大鼠, 工程化细胞

Abstract: BACKGROUND: Previous studies have shown that Huosui Formula has a synergistic effect on the immune and hematopoietic regulation of patients with myelodysplastic syndrome, but the specific mechanism is not yet clear.
OBJECTIVE: To explore the effect and mechanism of Huosui Formula on bone marrow hematopoiesis in rats with myelodysplastic syndrome.  
METHODS: A total of 70 SD rats were randomly divided into a normal control group (n=10), a model group (n=15), a western medicine group (n=15), a low-dose Huosui Formula group (n=15), and a high-dose Huosui Formula group (n=15). Except for the normal control group, the other four groups were injected with dimethylbenzanthracene via the tail vein to induce the establishment of rat myelodysplastic syndrome models. After modeling, the normal control group and the model group were given normal saline; the western medicine group was given thalidomide capsules 10 mg/kg and retinoic acid tablets 4 mg/kg, and the low-dose Huosui Formula group and the high-dose Huosui Formula group were given 1.5 and 6 g/kg Huosui Formula, respectively, by intragastric administration once a day for 28 consecutive days. Peripheral blood and femoral bone marrow tissue were collected to detect peripheral blood routine and bone marrow biopsy hematopoietic proliferation. Flow cytometry was used to detect T lymphocyte subsets and the expression of CTLA-4 and PD-1 on T lymphocytes. 
RESULTS AND CONCLUSION: (1) Compared with the normal control group, peripheral blood leukocyte, neutrophil, hemoglobin, platelet, and CD4+, CD4+/CD8+ levels were decreased in the model group significantly (P < 0.05), while CD4+PD-1+, CD8+PD-1+, CD4+CTLA-4+, and CD8+CTLA-4+ expressions were significantly upregulated (P < 0.05). (2) In all dosage groups, myelopoietic proliferation was increased compared with the model group, with no significant difference between the groups (P > 0.05). (3) Compared with the model group, leukocytes, hemoglobin, platelets, and CD4+, CD4+/CD8+ were significantly elevated in the high-dose Huosui Formula group (P < 0.05), the expression of CD8+ was significantly lower (P < 0.05), and the levels of CD4+PD-1+, CD8+PD-1+, CD4+CTLA-4+, and CD8+CTLA-4+ were down-regulated but not statistically significant (P > 0.05). (4) The western medicine group and the high-dose Huosui Formula group showed similar efficacy. The improvement of each index in the high-dose Huosui Formula group was superior to that in the low-dose Huosui Formula group. These findings indicate that Huosui Formula can improve the bone marrow hematopoiesis in myelodysplastic syndrome model rats, increase the levels of CD4+, and CD4+/CD8+ while down-regulate the expression levels of CD4+PD-1+, CD8+PD-1+, CD4+CTLA-4+, and CD8+CTLA-4+. These observations suggest a link to the negative immunoregulation mechanism. 

Key words: myelodysplastic syndrome, Huosui formula, bone marrow hematopoiesis, immune checkpoint, peripheral blood T lymphocyte subset, CTLA-4, PD-1, SD rat, engineered cell

中图分类号: