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黄长盛,辛兴涛,贺守第,谭宁.槲皮素对自然流产小鼠模型母胎免疫耐受的影响[J].湖南中医药大学学报英文版,2023,43(2):197-202.[Click to copy
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| 槲皮素对自然流产小鼠模型母胎免疫耐受的影响 |
| 黄长盛,辛兴涛,贺守第,谭宁 |
| (华中科技大学协和深圳医院中医风湿科, 广东 深圳 518000;河北省第八人民医院普外科, 河北 石家庄 050011) |
| 摘要: |
| 目的 研究槲皮素对自然流产（spontaneous abortion, SA）动物模型的影响，从免疫耐受调节的角度探讨槲皮素对SA的免疫机制。方法 将雌性CBA/J小鼠与雄性BALB/c小鼠合笼交配建立正常妊娠模型，为正常组，共6只；将雌性CBA/J小鼠与雄性DBA/2小鼠合笼交配建立SA模型，24只SA模型小鼠再随机分为模型组、地屈孕酮组、低浓度槲皮素组、高浓度槲皮素组，每组6只。第1天起，地屈孕酮组[3.75 mg/（kg·d）]、低浓度槲皮素组[300 mg/（kg·d）]、高浓度槲皮素组[600 mg/（kg·d）]开始灌胃，正常组、模型组灌胃等体积的助溶剂，1次/d。第14天，脱颈处死小鼠，计算孕鼠的流产率；ELISA法检测血清小鼠肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）、白细胞介素-10（interleukin-10, IL-10）、γ-干扰素（interferon-γ, IFN-γ）、雌二醇（estradiol, E2）、孕酮（progesterone, P）的水平；流式细胞术检测外周血CD3-CD56+CD16+自然杀伤（natural killer, NK）细胞、CD19+CD5+B细胞、CD4+T细胞、CD8+T细胞比例；HE染色观察子宫蜕膜组织结构。结果 与正常组比较，模型组流产率及TNF-α、IFN-γ水平升高（P<0.01），外周血CD3-CD56+CD16+NK细胞、CD19+CD5+B细胞、CD4+T细胞比例升高（P<0.01）；IL-10、E2、P水平和CD8+T细胞比例下降（P<0.01）；子宫蜕膜组织结构紊乱，胞浆空泡变增加，血管数量减少。与模型组比较，地屈孕酮组、低浓度槲皮素组、高浓度槲皮素组流产率及TNF-α、IFN-γ水平下降（P<0.01），外周血CD3-CD56+CD16+NK细胞、CD19+CD5+B细胞、CD4+T细胞比例下降（P<0.01），IL-10、E2、P水平和CD8+T细胞比例上升（P<0.01）；低浓度槲皮素组、高浓度槲皮素组的子宫蜕膜组织结构清晰，血管数量增加，组织间质水肿减少、胞浆空泡变减少。与地屈孕酮组比较，低浓度槲皮素组、高浓度槲皮素组流产率及IL-10、P水平明显升高（P<0.01），CD8+T细胞明显降低（P<0.01）；子宫蜕膜组织结构稍紊乱，胞浆空泡变少量增加，血管数量减少。结论 槲皮素能改善SA动物模型的流产率及蜕膜组织结构，可能与调节NK细胞、T细胞、B细胞及Th1/Th2细胞因子平衡有关。 |
| 关键词: 槲皮素 自然流产 免疫耐受 小鼠 NK细胞 Th1/Th2细胞因子 |
| DOI：10.3969/j.issn.1674-070X.2023.02.004 |
| Received:May 02, 2022 |
| 基金项目:广东省中医药管理局项目（20181235）；广东省省级科技计划项目公益研究与能力建设（2017A020215129）；深圳市科技计划项目基础研究（自由探索）（JCYJ20160429183052202）；深圳市南山区科技创新局项目（2020088，2020100）；华中科技大学协和深圳医院院内课题（2020007）。 |
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| Effects of quercetin on maternal-fetal immune tolerance in the spontaneous abortion mouse model |
| HUANG Changsheng,XIN Xingtao,HE Shoudi,TAN Ning |
| (Rheumatology Department of Chinese Medicine, Huazhong University of Science and Technology Union Shenzhen Hospital, Shenzhen, Guangdong 518000, China;Department of General Surgery, The Eighth People's Hospital of Hebei Province, Shijiazhuang, Hebei 050011, China) |
| Abstract: |
| Objective To study the effects of quercetin on spontaneous abortion (SA) animal model, and to further study the immune mechanism of quercetin on SA from the perspective of immune tolerance regulation. Methods Female CBA/J and male BALB/c mice were co-mated to establish the normal group with 6 mice, and female CBA/J mice were co-mated with male DBA/2 mice to establish SA model. Twenty-four SA model mice were randomly divided into normal group, model group, dydrogesterone group, low-concentration quercetin group, and high-concentration quercetin group, with 6 mice in each group. The mice in the dydrogesterone group were given intragastric administration of dydrogesterone with a dose of 3.75 mg/(kg·d), the mice in the low-concentration quercetin group and the high-concentration quercetin group were respectively given intragastric administration of quercetin with a dose of 300 mg/(kg·d) and 600 mg/(kg·d), the mice in the normal group and the model group were intragastrically administered an equal volume of cosolvent, once a day. On the 14th day, the mice were sacrificed by cervical dislocation to calculate the abortion rate of pregnant mice; ELISA method was used to detect the concentration of tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), gamma-interferon (IFN-γ), estradiol (E2) and progesterone (P) in serum of mice; flow cytometry was taken to detect the proportion of peripheral blood CD3-CD56+CD16+natural killer (NK) cells, CD19+CD5+B cells, CD4+T cells and CD8+T cells; the structure of uterine decidua was observed by HE staining. Results Compared with the normal group, in the model group, the abortion rate, the concentration of TNF-α, IFN-γ, and the proportion of peripheral blood NK cells, CD19+CD5+B cells, CD4+T cells increased (P<0.01); the concentration of IL-10, E2, P and the proportion of CD8+T cells were reduced (P<0.01); the model group was characterized by the structurely disordered uterine decidua, increased cytoplasmic vacuoles, and less blood vessels. Compared with the model group, the abortion rate, the concentration of TNF-α and IFN-γ, the proportion of peripheral blood CD3-CD56+CD16+NK cells, CD19+CD5+B cells, and CD4+T cells in the dydrogesterone group, low-concentration quercetin group and high-concentration quercetin group were reduced (P<0.01), while the concentration of IL-10, E2, P and the proportion of CD8+T cells increased (P<0.01); in the low-concentration quercetin group and high-concentration quercetin group, the structure of uterine decidua was clear, the number of blood vessels increased, the interstitial edema and the cytoplasmic vacuoles were reduced. Compared with the dydrogesterone group, in the low-concentration quercetin group and the high-concentration quercetin group, the abortion rate and the concentration of IL-10 and P were significantly higher (P<0.01), CD8+T cells decreased significantly (P<0.01), the structure of uterine decidua was slightly disordered, the cytoplasmic vacuoles increased slightly, and the number of blood vessels was reduced. Conclusion Quercetin can reduce the abortion rate and improve the structure of uterine decidua of SA animal model, which may be related to regulating the balance of NK cells, T cells, B cells and Th1/Th2 cytokines. |
| Key words: quercetin spontaneous abortion immune tolerance mice natural killer cell Th1/Th2 cytokine |
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