基于HPTT轴探讨补脾强力复方调节EAMG大鼠免疫系统平衡的机制

doi:10.11958/20220260

天津医药 ›› 2022, Vol. 50 ›› Issue (10): 1031-1036.doi: 10.11958/20220260

基于HPTT轴探讨补脾强力复方调节EAMG大鼠免疫系统平衡的机制

王强(), 况时祥, 李若照, 娄金波, 钱忆家, 雍波, 刘运权()

贵州中医药大学第二临床医学院（邮编550002）

收稿日期:2022-02-20 修回日期:2022-06-12 出版日期:2022-10-15 发布日期:2022-10-20
通讯作者: 刘运权 E-mail:liukangjing2006@163.com;1710547138@qq.com
作者简介:王强（1975）,女,副教授,主要从事中西医结合神经内科方面研究。E-mail： liukangjing2006@163.com
基金资助:
国家自然科学地区级基金资助项目(81960856);贵州省科学技术基金(黔科合J字[2015]2022号);贵州省教育厅青年科技人才成长项目(黔教合KY[2017]178)

Exploration on the regulation mechanism of Bupi Qiangli compound in immune system balance of EAMG rats based on HPTT axis

WANG Qiang(), KUANG Shixiang, LI Ruozhao, LOU Jinbo, QIAN Yijia, YONG Bo, LIU Yunquan()

The Second Clinical Medical College, Guizhou University of Traditional Chinese Medicine, Guiyang 550002, China

Received:2022-02-20 Revised:2022-06-12 Published:2022-10-15 Online:2022-10-20
Contact: LIU Yunquan E-mail:liukangjing2006@163.com;1710547138@qq.com

王强, 况时祥, 李若照, 娄金波, 钱忆家, 雍波, 刘运权. 基于HPTT轴探讨补脾强力复方调节EAMG大鼠免疫系统平衡的机制[J]. 天津医药, 2022, 50(10): 1031-1036.

WANG Qiang, KUANG Shixiang, LI Ruozhao, LOU Jinbo, QIAN Yijia, YONG Bo, LIU Yunquan. Exploration on the regulation mechanism of Bupi Qiangli compound in immune system balance of EAMG rats based on HPTT axis[J]. Tianjin Medical Journal, 2022, 50(10): 1031-1036.

摘要/Abstract

摘要：

目的基于下丘脑-垂体-甲状腺-胸腺（HPTT）轴探讨补脾强力复方调节实验性自身免疫性重症肌无力（EAMG）大鼠免疫系统平衡的机制。方法将120只Lewis大鼠随机分为对照组,模型组,补脾强力复方低、中、高剂量组（4.75、7.12、9.49 g/kg）和西药组（泼尼松,5.4 mg/kg）,每组20只。实验期间观察大鼠行为学改变（体质量、Lennon评分）。实验结束后HE染色评估胸腺组织病理变化,酶联免疫吸附试验（ELISA）检测血清乙酰胆碱受体抗体（AchR-Ab）、肿瘤坏死因子（TNF）-β、白细胞介素（IL）-4和IL-10含量;流式细胞术检测大鼠胸腺中CD4⁺CD25⁺Foxp3⁺ Treg细胞比例,同时检测HPTT轴功能损伤指标[（三碘甲状腺原氨酸（T3）、甲状腺素（T4）、促甲状腺激素（TSH）、促甲状腺激素释放激素（TRH）、下丘脑T3受体（T3R）、去甲肾上腺素（NE）]水平。结果建模后,与对照组相比,各建模组体质量下降,Lennon评分升高;给药后,补脾强力复方中、高剂量组和西药组体质量较模型组升高,Lennon评分下降（P<0.05）。HE染色可见模型组皮质区淋巴细胞变性坏死,髓质区胸腺小体消失,细胞数量减少;补脾强力复方低、中、高剂量组及西药组大鼠胸腺皮质区和髓质区细胞数量明显增多。与对照组相比,模型组胸腺CD4⁺CD25⁺Treg、CD4⁺CD25⁺Foxp3⁺ Treg细胞比例下降,CD4⁺CD25^-Foxp3⁺ Treg和CD4⁺CD25⁺Foxp3^- Treg细胞比例升高（P<0.05）,血清AchR-Ab、TNF-β、IL-4、IL-10、TSH和TRH水平明显升高,T3R和NE水平明显下降（P<0.05）。与模型组相比,补脾强力复方中、高剂量组及西药组胸腺CD4⁺CD25⁺Treg、CD4⁺CD25⁺Foxp3⁺ Treg细胞比例升高,CD4⁺CD25^-Foxp3⁺ Treg和CD4⁺CD25⁺Foxp3^- Treg细胞比例下降,血清AchR-Ab、TNF-β、IL-4、IL-10、TSH和TRH水平下降（P<0.05）;T3R和NE水平升高。结论补脾强力复方可以通过调控HPTT轴及其相关因子,改善EAMG大鼠自身免疫平衡。

关键词: 重症肌无力,自身免疫性,实验性, 补脾强力复方, 下丘脑-垂体-甲状腺-胸腺轴, 乙酰胆碱受体抗体, 调节性T细胞

Abstract:

Objective To investigate the mechanism of Bupi Qiangli compound in regulating immune system balance in experimental autoimmune myasthenia gravis (EAMG) rats based on the hypothalamic-pituitary-thyroid-thymus (HPTT) axis. Methods A total of 120 Lewis rats were randomly divided into the control group, the model group, the Bupi Qiangli compound groups (low-dose, medium-dose and high-dose groups, 4.75, 7.12 and 9.49 g/kg) and western medicine group (prednisone, 5.4 mg/kg), with 20 rats in each group. Behavioral changes (body weight, Lennon score) were observed during the experiment. After the experiment, histopathological changes of thymus were assessed by HE staining. The serum contents of acetylcholine receptor antibody (AchR-Ab), tumor necrosis factor-β (TNF-β), interleukin-4 (IL-4) and IL-10 were determined by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was used to detect the expression of CD4⁺CD25⁺Foxp3⁺Treg cells in rat thymus. At the same time, HPTT axis function damage index [triiodothyronine (T3), thyroxine (T4), TSH, thyroid stimulating hormone (TSH), thyroid stimulating hormone releasing hormone (TRH), hypothalamic T3 receptor (T3R) and norepineadrenalin (NE)] levels were detected. Results After modeling, compared with the control group, the body quality of each modeling group decreased and the Lennon score increased. After administration, body weights were increased in the Bupi Qiangli compound low-dose, medium-dose and high-dose groups and western medicine group than those of the model group, while values of Lennon score were decreased (P<0.05). HE staining showed degeneration and necrosis of lymphocytes in cortical area, disappearance of thymic bodies in medullary area, and decreased cell number in the model group. The number of cells in thymus cortical area and medulla area of rats were significantly increased in the low, medium and high dose groups of Bupi Qiangli compound and western medicine groups. Compared with the control group, the proportion of CD4⁺CD25⁺ Treg and CD4⁺CD25⁺Foxp3⁺ Treg in thymus were decreased in the model group, and the proportion of CD4⁺CD25^-Foxp3⁺ Treg and CD4⁺CD25⁺Foxp3⁺ Treg were increased (P<0.05). The serum levels of AchR-Ab, TNF-β, IL-4, IL-10, TSH and TRH were significantly increased, while the levels of T3R and NE were significantly decreased (P<0.05). Compared with the model group, the proportion of CD4⁺CD25⁺ Treg and CD4⁺CD25⁺Foxp3⁺ Treg were significantly increased in the Bupi Qiangli compound medium-dose and high-dose groups and western medicine group, while the proportion of CD4⁺CD25^-Foxp3⁺ Treg and CD4⁺CD25⁺Foxp3⁺ Treg decreased. Serum AchR-Ab, TNF-β, IL-4, IL-10, TSH and TRH levels were significantly decreased (P<0.05). T3R and NE levels were increased. Conclusion Bupi Qiangli compound can improve the autoimmune balance of EAMG rats by regulating HPTT axis and related factors.

Key words: hypothalamus-pituitary-thyroid-thymus, Bupi Qiangli compound, experimental autoimmune myasthenia gravis, receptor antibody, regulatory T cell

中图分类号:

R746.1

图/表 6

参考文献 23

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组别	建模前	建模后	给药后
对照组	190.33±1.73	278.08±2.63	303.92±2.49
模型组	191.14±2.02	205.21±5.29^a	193.88±2.86^a
低剂量组	189.69±1.46	201.44±3.00^a	208.63±3.04^a
中剂量组	190.72±1.47	211.13±6.19^a	213.69±4.29^ab
高剂量组	190.38±1.78	209.66±5.46^a	227.15±2.94^ab
西药组	191.34±2.41	203.87±2.72^a	229.40±3.21^ab
F	2.117	928.100^＊＊	295.100^＊＊

组别	建模前	建模后	给药后
对照组	190.33±1.73	278.08±2.63	303.92±2.49
模型组	191.14±2.02	205.21±5.29^a	193.88±2.86^a
低剂量组	189.69±1.46	201.44±3.00^a	208.63±3.04^a
中剂量组	190.72±1.47	211.13±6.19^a	213.69±4.29^ab
高剂量组	190.38±1.78	209.66±5.46^a	227.15±2.94^ab
西药组	191.34±2.41	203.87±2.72^a	229.40±3.21^ab
F	2.117	928.100^＊＊	295.100^＊＊

组别	建模前	建模后	给药后
对照组	0	0	0
模型组	0	2.1±0.3^a	2.5±0.3^a
低剂量组	0	2.0±0.4^a	1.7±0.3^ab
中剂量组	0	1.9±0.5^a	1.6±0.3^ab
高剂量组	0	2.1±0.4^a	1.1±0.2^abcd
西药组	0	2.2±0.3^a	1.2±0.2^abcd
F	-	57.410^＊＊	117.100^＊＊

组别	建模前	建模后	给药后
对照组	0	0	0
模型组	0	2.1±0.3^a	2.5±0.3^a
低剂量组	0	2.0±0.4^a	1.7±0.3^ab
中剂量组	0	1.9±0.5^a	1.6±0.3^ab
高剂量组	0	2.1±0.4^a	1.1±0.2^abcd
西药组	0	2.2±0.3^a	1.2±0.2^abcd
F	-	57.410^＊＊	117.100^＊＊

组别		CD4⁺CD25⁺		CD4⁺CD25⁺Foxp3⁺
对照组		70.13±12.42		75.58±12.64
模型组		56.92±10.04^a		49.21±6.02^a
低剂量组		61.82±11.78		58.87±12.46^a
中剂量组		67.99±14.52^b		63.77±13.40^ab
高剂量组		69.95±15.67^bc		71.24±12.78^bcd
西药组		70.03±13.84^bc		75.13±15.81^bcd
F		13.761^＊＊		6.857^＊＊
组别	CD4⁺Foxp3⁺		CD4⁺CD25^-Foxp3⁺		CD4⁺CD25⁺Foxp3^-
对照组	98.61±4.69		27.19±9.63		2.59±0.96
模型组	95.48±3.59		45.12±4.11^a		4.18±0.62^a
低剂量组	95.91±4.38		38.44±3.03^a		3.52±0.73
中剂量组	96.42±3.25		30.18±4.19^bc		3.25±0.69^b
高剂量组	97.16±4.08		29.78±3.46^bc		2.83±0.57^b
西药组	95.27±3.05		28.09±2.72^bc		2.79±0.80^b
F	1.032		19.461^＊＊		6.374^＊＊

基于HPTT轴探讨补脾强力复方调节EAMG大鼠免疫系统平衡的机制

Exploration on the regulation mechanism of Bupi Qiangli compound in immune system balance of EAMG rats based on HPTT axis

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组别	AchR-Ab（pmol/L）	TNF-β（ng/L）	IL-4（ng/L）	IL-10（ng/L）
对照组	0.23±0.05	58.26±6.69	42.13±9.73	45.23±7.52
模型组	0.45±0.04^a	89.18±9.59^a	68.83±15.83^a	72.11±8.02^a
低剂量组	0.41±0.06^a	77.49±10.04^ab	61.93±14.52^a	68.37±9.46^a
中剂量组	0.38±0.05^ab	69.42±8.28^abc	58.24±12.34^ab	61.37±8.47^ab
高剂量组	0.26±0.08^bcd	61.06±7.35^bcd	49.39±11.53^b	50.14±6.78^bcd
西药组	0.24±0.04^bcd	59.56±6.57^bcd	48.27±10.94^b	48.63±7.41^bcd
F	30.516^＊＊	22.223^＊＊	6.143^＊＊	19.736^＊＊

组别	T3（μg/L）	T4（μg/L）	T3R（fmol/mg DNA）	NE（ng/g）	TSH（μg/L）	TRH（ng/L）
对照组	1.92±0.26	119.41±9.49	178.51±32.63	793.19±122.96	11.39±1.66	154.61±31.61
模型组	1.87±0.19	112.05±10.43	125.32±45.29^a	453.59±102.86^a	29.94±3.38^a	289.82±51.59^a
低剂量组	1.94±0.21	116.23±10.13	131.40±39.96^a	508.96±113.04^a	26.61±2.34^a	227.19±51.42^ab
中剂量组	1.88±0.10	111.89±17.06	155.41±36.19^ab	553.27±104.39^ab	21.65±1.18^bc	195.26±45.26^b
高剂量组	1.93±0.25	110.99±11.55	169.57±45.46^bcd	687.22±122.34^bcd	18.92±1.23^abcd	161.86±44.37^bc
西药组	1.82±0.07	111.42±16.22	173.99±42.72^bcd	696.24±123.31^bcd	19.82±1.40^abcd	159.23±35.75^bc
F	0.551	0.698	3.100^＊	12.774^＊＊	102.272^＊＊	14.465^＊＊

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