针康法对脑缺血小鼠神经功能及叉头转录因子3和维甲酸相关孤儿受体γt表达的影响

doi:10.3969/j.issn.1006-9771.2021.12.009

《中国康复理论与实践》 ›› 2021, Vol. 27 ›› Issue (12): 1437-1442.doi: 10.3969/j.issn.1006-9771.2021.12.009

针康法对脑缺血小鼠神经功能及叉头转录因子3和维甲酸相关孤儿受体γt表达的影响

杨赫¹,唐强²(),朱路文¹

1. 黑龙江中医药大学,黑龙江哈尔滨市 150040
2. 黑龙江中医药大学附属第二医院,黑龙江哈尔滨市150001

收稿日期:2021-07-08 修回日期:2021-10-25 出版日期:2021-12-25 发布日期:2021-12-29
通讯作者: 唐强 E-mail:tangqiang1963@163.com
作者简介:杨赫(1987-),男,汉族,黑龙江哈尔滨市人,博士研究生,主要研究方向：脑卒中的中医康复基础研究。
基金资助:
1.黑龙江省自然科学基金重点项目(ZD2019H007);2.黑龙江中医药大学科研基金项目(2019MS22)

Effect of Acupuncture-rehabilitation Therapy on Neural Function and Expression of Forkhead Box P3 and Retinoic Acid-related Orphan Receptor γt in Mice with Cerebral Ischemia

YANG He¹,TANG Qiang²(),ZHU Lu-wen¹

1. Heilongjiang University of Chinese Medicine, Harbin, Heilongjiang 150040, China
2. The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin, Heilongjiang 150001, China

Received:2021-07-08 Revised:2021-10-25 Published:2021-12-25 Online:2021-12-29
Contact: TANG Qiang E-mail:tangqiang1963@163.com
Supported by:
Heilongjiang Natural Science Foundation(ZD2019H007);Heilongjiang University of Chinese Medicine Scientific Research Fund(2019MS22)

摘要/Abstract

摘要：

目的探索针康法对脑缺血小鼠叉头转录因子3 (Foxp3)和维甲酸相关孤儿受体γt (RORγt)蛋白表达的影响。方法 45只成年雌性C57BL/6小鼠随机分为假手术组、模型组、针刺组、康复组和针康组,每组9只。每组再分为3 d、7 d、14 d 3个亚组。模型组、针刺组、康复组和针康组采用线栓法建立永久性大脑中动脉栓塞模型。假手术组和模型组仅手术不予治疗,针刺组接受头穴丛刺针灸治疗,康复组接受跑台训练,针康组接受头穴丛刺结合跑台治疗。术后各时间点,采用改良神经功能缺损评分(mNSS)评价神经功能,Western blotting法分析缺血侧脑组织Foxp3、RORγt表达。结果术后各时间点,假手术组mNSS为0分,模型组mNSS均高于假手术组(P < 0.05)。术后3 d,与模型组比较,康复组、针康组mNSS降低(P < 0.05)。术后14 d,与模型组和针刺组比较,针康组mNSS降低(P < 0.05)。术后各时间点,针康组Foxp3蛋白表达均低于模型组(P < 0.05)。术后3 d,针康组RORγt表达量较其他组均增加(P < 0.05)。术后7 d,针康组RORγt表达量较针刺组、假手术组增加(P < 0.05)。结论针康法可改善脑缺血小鼠组织损伤,促进神经功能恢复,可能通过调节Foxp3、RORγt表达降低炎症的水平,发挥神经保护作用。

关键词: 脑缺血, 针康法, 叉头转录因子3, 维甲酸相关孤儿受体γt;, 小鼠

Abstract:

Objective To explore the effect of acupuncture-rehabilitation therapy on the expression of transcription factor forkhead box P3 (Foxp3) and retinoic acid-related orphan receptor γt (RORγt) protein in cerebral ischemic mice. Methods Forty-five female C57BL/6 mice were randomly assigned to sham operation group, model group, acupuncture group, rehabilitation group, and acupuncture-rehabilitation group, with nine mice in each group. Subsequently, each group was divided into three days, seven days and 14 days subgroups. The permanent middle cerebral artery occlusion models were established by the suture method, except the sham operation group. The sham operation group and the model group received no treatment. The acupuncture group received scalp cluster acupuncture, the rehabilitation group received treadmill training, and the acupuncture-rehabilitation group received scalp cluster acupuncture combined with treadmill training. Three days, seven days and 14 days after modeling, the modified Neurological Severity Score (mNSS) was obtained, and the expression of Foxp3 and RORγt in brain tissue of ischemic side was analyzed by Western blotting. Results The mNSS in the sham operation group was 0, and was higher in the model group than in the sham operation group at each postoperative time point. Three days after operation, the mNSS decreased in the rehabilitation group and the acupuncture-rehabilitation group, compared to the model group (P < 0.05). Fourteen days after operation, the mNSS decreased in the acupuncture-rehabilitation group, compared to the model group and acupuncture group (P < 0.05). The expression of Foxp3 protein was significantly lower in the acupuncture-rehabilitation group than in other groups at all time points after surgery (P < 0.05). Three days after operation, the expression of RORγt was higher in the acupuncture-rehabilitation group than in other groups (P < 0.05). Seven days after operation, the expression of RORγt was higher in the acupuncture-rehabilitation group than in the acupuncture group and sham operation group (P < 0.05). Conclusion Acupuncture-rehabilitation therapy may improve the tissue injury of cerebral ischemia mice, and promote the recovery of neural function, possibly by regulating Foxp3 and RORγT expression to reduce the level of inflammation, and then exert neuroprotective effects.

Key words: cerebral ischemia, acupuncture-rehabilitation therapy, Forkhead box P3, retinoid-related orphan nuclear receptor γt;, mice

中图分类号:

R743.3

杨赫,唐强,朱路文. 针康法对脑缺血小鼠神经功能及叉头转录因子3和维甲酸相关孤儿受体γt表达的影响[J]. 《中国康复理论与实践》, 2021, 27(12): 1437-1442.

YANG He,TANG Qiang,ZHU Lu-wen. Effect of Acupuncture-rehabilitation Therapy on Neural Function and Expression of Forkhead Box P3 and Retinoic Acid-related Orphan Receptor γt in Mice with Cerebral Ischemia[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(12): 1437-1442.

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组别	n	3 d	7 d	14 d	F值	P值
假手术组	3	0.00±0.00	0.00±0.00	0.00±0.00
模型组	3	11.67±0.58^a	9.00±1.40^a	6.33±0.58^a	8.123	0.020
针刺组	3	8.33±0.58^a	6.67±0.58^a	5.33±0.58^a	6.714	0.029
康复组	3	5.67±2.08^a,b	3.67±1.15	4.03±1.00^a	0.513	0.623
针康组	3	5.67±1.53^a,b	5.33±2.73	2.33±0.57^a,b,c	1.002	0.421
F值		12.506	5.153	15.770
P值		< 0.001	0.016	< 0.001

组别	n	3 d	7 d	14 d	F值	P值
假手术组	3	1.00±0.00	1.00±0.00	1.00±0.00
模型组	3	1.00±0.05	0.80±0.11	0.91±0.06	1.654	0.268
针刺组	3	0.81±0.11	0.74±0.12	0.81±0.04	0.174	0.844
康复组	3	0.72±0.07	0.70±0.09	0.79±0.09	0.318	0.739
针康组	3	0.54±0.08^a,b	0.24±0.16^a,b,c,d	0.66±0.05^a,b	4.070	0.076
F值		7.390	6.502	5.231
P值		0.005	0.008	0.015

组别	n	3 d	7 d	14 d	F值	P值
假手术组	3	1.00±0.00	1.00±0.00	1.00±0.00
模型组	3	1.02±0.13	1.22±0.16	0.82±0.04^a	2.721	0.144
针刺组	3	0.55±0.33	1.02±0.13	0.89±0.02^a	1.400	0.317
康复组	3	0.57±0.35	1.46±0.13	1.05±0.02^b,c	4.258	0.071
针康组	3	2.54±0.29^a,b,c,d	1.55±0.10^a,c	0.87±0.04^a,d	22.108	0.002
F值		10.029	4.510	11.413
P值		0.002	0.024	< 0.001

针康法对脑缺血小鼠神经功能及叉头转录因子3和维甲酸相关孤儿受体γt表达的影响

Effect of Acupuncture-rehabilitation Therapy on Neural Function and Expression of Forkhead Box P3 and Retinoic Acid-related Orphan Receptor γt in Mice with Cerebral Ischemia

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