左归丸和右归丸对实验性自身免疫性脑脊髓炎大鼠中枢神经系统IL-10、TGF-β蛋白表达的研究

首都医科大学学报 ›› 2010, Vol. 31 ›› Issue (2): 233-240.

左归丸和右归丸对实验性自身免疫性脑脊髓炎大鼠中枢神经系统IL-10、TGF-β蛋白表达的研究

樊永平^1*, 宋丽君¹, 叶明¹, 周莉¹, 王蕾², 龚海洋¹

1. 首都医科大学附属北京天坛医院中医科;2. 首都医科大学中医药学院方剂学教研室

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-04-21 发布日期:2010-04-21
通讯作者: 樊永平

Effects of ZuoGuiWan and YouGuiWan on the Expression of IL-10 and TGF-β in Central Nervous System of Experimental Autoimmune Encephalomyelitis Rats

FAN Yong-ping^1*, SONG Li-jun¹, YE Ming¹, ZHOU Li¹, WANG Lei², GONG Hai-yang¹

1. Department of Traditional Chinese Medicine, Beijing Tiantan Hospital, Capital Medical University; 2. Department of Prescription, School of Traditional Chinese Medicine, Capital Medical University

Received:1900-01-01 Revised:1900-01-01 Online:2010-04-21 Published:2010-04-21
Contact: FAN Yong-ping

摘要/Abstract

摘要： 目的观察大鼠实验性自身免疫性脑脊髓炎（experimental autoimmune encephalomyelitis，EAE）模型脑、脊髓组织白细胞介素-10（interleukin-10，IL-10）与组织生长因子-β（transforming growth factor-β，TGF-β）免疫组化的表达，探讨滋阴补肾法（左归丸）与温阳补肾法（右归丸）治疗EAE的作用机制。方法用髓鞘碱性蛋白（myelin basic protein，MBP）免疫Lewis大鼠诱导EAE模型，120只大鼠采用数字表法随机分为正常组、模型组、激素组、左归丸组及右归丸组，正常对照组（n=24）和模型组（n=24）每天给予3 mL 0.9%氯化钠注射液灌胃；激素组（n=24）免疫后每天给予3 mL 0.9%氯化钠注射液灌胃，发病后改为醋酸泼尼松混悬液5 mg/(kg·d^-1)灌胃；左归丸组（n=24）免疫后给予左归丸混悬液2 g/(kg·d^-1)；右归丸组（n=24）免疫后给予右归丸混悬液3 g/(kg·d^-1)，直至处死。分别于免疫后15 d（急性期发病）和免疫后27 d（缓解期）选取各组大鼠，取脑和脊髓组织切片进行IL-10、TGF-β免疫组化染色。结果造模后15 d，模型动物的脑和脊髓组织IL-10、TGF-β表达（指表达染色IOD值，下同）明显减少，第27天，模型动物脊髓组织IL-10表达明显减少。第15天，左归丸组与右归丸组大鼠脑组织IL-10表达较模型组明显升高（P<0.01）；左归丸组大鼠脊髓组织IL-10表达也较模型组明显升高（P<0.05）；右归丸组与激素组大鼠脑TGF-β表达较模型组升高，右归丸组大鼠脊髓TGF-β表达较模型组升高，差异均有统计学意义（P<0.05）。造模后27 d，左归丸组、右归丸组、激素组脊髓组织IL10表达均较模型组显著增高（P<0.01）；左归丸组大鼠脊髓组织TGF-β表达较模型组与激素组升高（P<0.05）。模型组、激素组、左归丸组与右归丸组脊髓IL-10表达均较免疫后15 d明显升高（P<0.05）；左归丸组大鼠脊髓TGF-β表达较免疫后15 d明显增高，差异有统计学意义（P<0.05）。结论左归丸和右归丸可上调EAE大鼠中枢神经系统IL-10、TGF-β表达，作用优于激素。两者整个病程中均对IL-10表达有促进作用，不同点为右归丸促进急性期脑组织TGF-β的表达，而左归丸则促进缓解期脊髓TGF-β的表达。

关键词: 自身免疫性疾病, 实验性自身免疫性脑脊髓炎, 细胞因子, 中医药, 左归丸, 右归丸

Abstract: Objective To observe the immunohistochemical expression of interleukin-10(IL-10), transforming growth factor-β(TGF-β) in brain and spinal cord tissue, and explore the mechanism of nourishing kidney-Yin（Zuo Gui Wan） and invigorating kidney-Yang（You Gui Wan）in rats with experimental autoimmune encephalomyelitis(EAE). Methods Lewis rats were immunized with the myelin basic protein(MBP). Totally 120 Rats were randomly divided into Normal-group, EAE-group, prednisone-group, Zuo Gui Wan group and You Gui Wan group after immunization with 24 rats in each. Rats in normal-group and EAE-group were given normal saline by gastric gavage, 3 mL/d; rats in prednisone-group were treated with suspension of prednisone after clinical signs developed, 5 mg/(kg·d^-1); rats in Zuo Gui Wan group were treated with suspension of Zuo Gui Wan, 2 g/(kg·d^-1), and rats in You Gui Wan group were treated with suspension of You Gui Wan, 3 g/(kg·d^-1). On days 15 and 27 post immune, the rats were sacrificed and the immunohistochemical staining was performed on the sections of brain and spinal cord. Results On day 15 post immune, the expression of IL-10 in brain tissue of Zuo Gui Wan group and You Gui Wan group were significantly higher(P<0.01) than that in EAE-group. The expression of IL-10 in spinal cord tissue of ZuoGuiWan group was significantly higher than that in EAE-group(P<0.05). The expression of TGF-β in brain tissue of You Gui Wan group and prednisone-group were significantly higher(P<0.05) than that in EAE-group. The expression of TGF-β in spinal cord tissue of You Gui Wan group was significantly higher than that in EAE-group(P<0.05). On day 27 post immune, expression of IL-10 in spinal cord tissue of prednisone-group, Zuo Gui Wan group and You Gui Wan group was significantly higher(P<0.01) than EAE-group. The expression of TGF-β in spinal cord tissue of Zuo Gui Wan group was significantly higher than that in EAE-group and prednisone-group(P<0.05). Expression of IL-10 in spinal cord tissue of EAE-group, prednisone-group, Zuo Gui Wan group and You Gui Wan group was significantly higher(P<0.05) than that on day 15 post immune, the expression of TGF-β in spinal cord tissue of Zuo Gui Wan group was higher than that on day 15 post immune(P<0.05). Conclusion Both Zuo Gui Wan and You Gui Wan have the effect of upregulating the immunohistochemical expression of IL-10 and TGF-β in central nervous system of EAE rats, promoting the recovery from the disease, their effects are better than those of prednisone. But You Gui Wan could only promote the expression of TGF-β in both brain and spinal cord tissue in acute stage(15 d) of EAE rats, and Zuo Gui Wan could promote the expression of TGF-β in spinal cord tissue in recovering stage(27 d).

Key words: autoimmune diseases, experimental autoimmune encephalomyelitis, cytokines, traditional Chinese medicine, Zuo Gui Wan, You Gui Wan

中图分类号:

R 744.3

樊永平;宋丽君;叶明;周莉;王蕾;龚海洋. 左归丸和右归丸对实验性自身免疫性脑脊髓炎大鼠中枢神经系统IL-10、TGF-β蛋白表达的研究[J]. 首都医科大学学报, 2010, 31(2): 233-240.

FAN Yong-ping;SONG Li-jun;YE Ming;ZHOU Li;WANG Lei;GONG Hai-yang. Effects of ZuoGuiWan and YouGuiWan on the Expression of IL-10 and TGF-β in Central Nervous System of Experimental Autoimmune Encephalomyelitis Rats[J]. Journal of Capital Medical University, 2010, 31(2): 233-240.

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