| 吴静泽,王晔恺,陈位,金丹雯.当归六黄汤通过激活Tyro3/Axl受体活性治疗桥本甲状腺炎大鼠的实验研究[J].浙江中西医结合杂志,2025,35(3): |
| 当归六黄汤通过激活Tyro3/Axl受体活性治疗桥本甲状腺炎大鼠的实验研究 |
| Effect of danggui liuhuang decoction on rat model with hashimoto"s thyroiditis by activation of Tyro3/Axl signal pathway |
| 投稿时间：2024-03-01 修订日期：2024-08-02 |
| DOI： |
| 中文关键词: 桥本甲状腺炎 当归六黄汤 Tyro3/Axl受体 B淋巴细胞瘤-2 Bcl-2相关X蛋白 |
| 英文关键词:Hashimoto's thyroiditis Danggui liuhuang decoction Tyro3/Axl receptor Bcl-2 Bax |
| 基金项目:浙江省中医药管理局科技计划项目(2023ZL768) |
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| 中文摘要: |
| 【目的】探讨当归六黄汤治疗桥本甲状腺炎大鼠的可能作用机制。【方法】24只大鼠适应性喂养1周后，按随机数字法分为空白组、模型组、安慰剂组及中药组，每组6只。空白组不予任何处理直接处死。模型组、安慰剂组和中药组大鼠通过将甲状腺球蛋白在尾根和足垫进行皮下注射各100 μg，每周1次，持续7周，建立桥本甲状腺炎模型。从第8周开始对安慰剂组和中药组分别进行40 g/L的安慰剂和144 g/L当归六黄汤水煎液灌胃2 mL，每天1次。在第11周处死模型组、安慰剂组和中药组所有大鼠。CT检测大鼠甲状腺组织肿胀情况。流式检测外周血CD3细胞表面Tyro3、Axl的平均荧光强度，酶联免疫吸附试验（ELISA）检测大鼠血清抗甲状腺球蛋白抗体（TG-Ab）和抗甲状腺过氧化物酶抗体（TPO-Ab）水平，免疫组化检测大鼠甲状腺组织Tyro3和Axl表达，实时荧光定量PCR检测甲状腺组织Tyro3、Axl、B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）mRNA表达。【结果】CT下可见中药组相对于模型组和安慰剂组，其甲状腺组织炎性减退，边界逐渐清晰。与空白组比较，模型组血清TG-Ab[ (79.77±6.06)U/mL比(43.00±9.36)U/mL，P＜0.01]、TPO-Ab[(8.04±0.99)U/mL比(2.76±1.11)U/mL，P＜0.01]及甲状腺组织Bax mRNA[(1.39±0.09)比(0.27±0.11)，P＜0.01]表达均升高，甲状腺组织Bcl-2 mRNA[ (1.29±0.14)比(3.39±0.37)，P＜0.01]表达降低。与模型组和安慰剂组比较，中药组血清TG-Ab[ (43.69±11.51)U/mL比(79.77±6.06)U/mL、(82.17±12.11)U/mL，P＜0.01]、TPO-Ab[ (3.02±0.73)U/mL比(8.04±0.99)U/mL、(7.84±0.79) U/mL，P＜0.01]及甲状腺组织Bax mRNA[ (0.24±0.10)比(1.39±0.09)、(1.38±0.34)，P＜0.01]表达均降低，甲状腺组织Bcl-2 mRNA表达升高[Bcl-2 mRNA: (2.89±0.32)比(1.29±0.14)、(1.36±0.29)，P＜0.01]。与空白组比较，模型组外周血CD3+T上Tyro3/Axl平均荧光强度[Tyro3(1325.53±129.45)比(1775.95±120.73)，Axl(130.10±11.71)比(156.58±9.27)，P＜0.01]及甲状腺组织Tyro3/Axl mRNA[Tyro3：(0.67±0.13)比(3.06±0.23)，Axl：(0.14±0.05)比(0.71±0.17)，P＜0.01]和平均光密度均降低[Tyro3：(119.45±14.47)比(266.20±33.21)，Axl： (117.96±13.06)比(240.06±21.50)，P＜0.01]。与模型组和安慰剂组比较，中药组外周血CD3+T上Tyro3/Axl平均荧光强度[Tyro3: (1769.92±151.90)比(1325.53±129.45)、(1241.57±193.40)，Axl: (150.08±15.32)比(130.10± 11.71)、(127.50±9.94)，P＜0.01]及甲状腺组织Tyro3/Axl mRNA[Tyro3：(2.69±0.44)比(0.67±0.13)、(0.71±0.33)，Axl： (0.62±0.10)比(0.14±0.05)、(0.12±0.04)，P＜0.01]和平均光密度[Tyro3：(251.41±32.28)比(119.45±14.47)、(115.69±24.81)，Axl：(223.23±19.12)比(117.96±13.06)、(119.69±20.48)，P＜0.01]均升高。【结论】当归六黄汤对桥本甲状腺炎的抑制效果可能和激活Tyro3/Axl受体信号通路进而引起凋亡抑制有关。 |
| 英文摘要: |
| Objective To invesigate the mechanism of danggui liuhuang decoction on rat model with hashimoto's thyroiditis. Method After 1 week of acclimation, 24 male SD rats were randomly divided into 4 groups:blank group(6 rats),model group(6 rats),placebo group(6 rats), traditional medicine group(6 rats).The blank group was directly executed for background testing. Hashimoto's thyroiditis model was established by injecting 100 μg thyroid globulin in tail root and foot pad respectively onec a week for 7 weeks, in model group, placebo group and traditional medicine group. From 8th week, placebo group and traditional medicine group were given 40 g/L placebo solution and 144 g/L water decoction of danggui liuhuang respectively, gavage administration with 2 ml once a day. In 11th week, blood and thyroid tissue of model group,placebo group and traditional medicine group were collected respectivelly. Swelling condition in thyroid tissue was detected by CT. Mean fluorescence intensity(MFI) of Tyro3 and Axl in CD3+T cells was detected by flow cytometry. Serum levels of TG-Ab and TPO-Ab were detected by enzyme-linked immunosorbent assay(ELISA). Tyro3 and Axl expression in thyroid tissue were detected by immunohistochemical. Tyro3, Axl, Bcl-2, Bax mRNA refer to β-actin mRNA were detected by Quantitative Real-time PCR. Results Compared to model group and placebo group, CT showed clearer boundaries of thyroid tissue in the traditional medicine group.Compared to blank group, TG-Ab(79.77±6.06 vs 43.00±9.36 U/ml, P<0.01), TPO-Ab(TPO-Ab : 8.04±0.99 vs 2.76±1.11 U/ml, P<0.01) levels in serum and Bax mRNA(Bax: 1.39±0.09 vs 0.27±0.11, P<0.01) in thyroid tissue in model group increased, Bcl-2 mRNA(1.29±0.14 vs 3.39±0.37, P<0.01) decreased.Compared to model and placebo group, TG-Ab(43.69±11.51 vs 79.77±6.06, 82.17±12.11 U/ml),TPO-Ab(3.02±0.73 vs 8.04±0.99,7.84±0.79 U/ml, P<0.01) levels in serum and Bax mRNA(0.24±0.10 vs1.39± 0.09,1.38±0.34, P<0.01) in thyroid tissue in traditional medicine group decreased,Bcl-2 mRNA(2.89±0.32 vs 1.29±0.14,1.36±0.29, P<0.01) increased.
Compared to blank group, Tyro3 MFI, Axl MFI(Tyro3: 1325.53 ±129.45 vs 1775.95±120.73, Axl: 130.10±11.71 vs 156.58±9.27,P<0.01) in CD3+T cells and Tyro3 mRNA, Axl mRNA(Tyro3: 0.67±0.13 vs 3.06±0.23，Axl: 0.14±0.05 vs 0.71±0.17),Tyro3 AOD, Axl AOD(Tyro3: 119.45±14.47 vs 266.20±33.21, Axl: 117.96±13.06 vs 240.06±21.50, P<0.01) in thyroid tissue in model group decreased.Compared to model group and placebo group, Tyro3 MFI, Axl MFI(Tyro3: 1769.92±151.90 vs 1325.53± 129.45,1241.57±193.40, Axl: 150.08±15.32 vs 130.10± 11.71, 127.50±9.94,P<0.01) in CD3+T cells and Tyro3 mRNA, Axl mRNA(Tyro3: 2.69±0.44 vs 0.67±0.13,0.71±0.33，Axl: 0.62±0.10 vs 0.14±0.05, 0.12±0.04,P<0.01) Tyro3 AOD, Axl AOD(Tyro3: 251.41±32.28 vs 119.45±14.47,115.69±24.81, Axl: 223.23± 19.12 vs 117.96±13.06, 119.69±20.48,P<0.01) in thyroid tissue in traditional medicine group increased.Conclusion Danggui liuhuang decoction effect on inhibiting apoptosis in hashimoto's thyroiditis by mechanism of activation of Tyro3/Axl receptor signal pathway. |
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