二妙方加细辛（北细辛）对骨关节炎大鼠模型骨髓干细胞向病灶区定向归巢的影响。

Effect of ermiao fang with xixin (herba asari mandshurici) on bone marrow stem cell directional homing to a focal zone in an osteoarthritis rat model.

出版信息

J Tradit Chin Med. 2014 Aug;34(4):477-87. doi: 10.1016/s0254-6272(15)30050-9.

Abstract

OBJECTIVE

To investigate the effects of Ermiao Fang (EM) with medical guide Xixin (Herba Asari Mandshurici) (HAM) on bone marrow stem cell migration to a focal zone in osteoarthritis (OA) rats.

METHODS

OA rats were induced by arthrectomy and assigned to sham-operated, model, EM, or EM plus HAM groups. All rats were injected with recombinant human granulocyte colony-stimulating factor 30 microg x kg(-1) x d(-1) for 7 days and treated with EM or EM plus HAM at 1.6 or 1.9 g x kg(-1) x d(-1) for 3 or 6 weeks, respectively. Chondrocyte apoptosis and cartilage matrix components were tested by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay and special staining. Levels of interleukin-1 beta (IL-1beta) tumor necrosis factor alpha (TNF-alpha) nitric oxide (NO), and inducible nitric oxide synthase (iNOS) in serum were detected by enzyme-linked immunosorbent assay or radioimmunoassay. Matrix metalloproteinases (MMPs)-13, tissue inhibitors of metalloproteinases (TIMPs)-1, Bromodeoxyuridine (BrdU), cluster of differentiation 34 (CD34), and stromal cell-derived factor 1 (SDF-1) were measured by immunohistochemical assay.

RESULTS

The EM and EM plus HAM groups had significantly less cartilage damage and synovium inflammation the model group. Moreover, the EM and EM plus HAM groups had less chondrocyte apoptosis and more proteoglycan and collagen content than the model group. The EM and EM plus HAM groups had obviously higher MMPs-13 and TIMPs-1 expression in the cartilage than the model group. Moreover, the two formula groups had less release of IL-1beta, TNF-alpha, NO, and iNOS than model group. Importantly, the expressions of BrdU, CD34, and SDF-1 in cartilage were significantly higher in the EM and EM plus HAM-Medtreated rats than model group. Notably, the EM plus HAM treatment seemed to have the greatest effects.

CONCLUSION

HAM improves the therapeutic effects of EM on OA rats by enhancing BMSC directional homing to the focal zone.

摘要

目的

观察细辛（Herba Asari Mandshurici）伍以二妙丸（Ermiao Fang）对骨关节炎（OA）大鼠骨髓间充质干细胞（BMSC）向病灶归巢的影响。

方法

采用关节切除法制备 OA 大鼠模型，随机分为假手术组、模型组、二妙丸组和二妙丸加细辛组。术后第 2 天各组大鼠均腹腔注射 rhG-CSF（30μg·kg-1·d-1），连续 7d。从第 3 天起，二妙丸组和二妙丸加细辛组分别给予二妙丸和二妙丸加细辛混悬液灌胃，1.6g·kg-1·d-1 和 1.9g·kg-1·d-1，连续 3 周或 6 周。采用末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记法（TUNEL）和特殊染色法检测软骨细胞凋亡，番红 O 固绿染色法检测软骨基质成分。酶联免疫吸附法（ELISA）和放射免疫法检测血清白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）、一氧化氮（NO）和诱导型一氧化氮合酶（iNOS）水平。免疫组化法检测基质金属蛋白酶-13（MMP-13）、基质金属蛋白酶组织抑制因子-1（TIMP-1）、BrdU、CD34 和基质细胞衍生因子-1（SDF-1）。

结果

与模型组相比，二妙丸组和二妙丸加细辛组关节软骨损伤和滑膜炎症明显减轻，软骨细胞凋亡减少，软骨基质中蛋白多糖和胶原含量增加。二妙丸组和二妙丸加细辛组关节软骨中 MMP-13 和 TIMP-1 表达均明显高于模型组。二妙丸组和二妙丸加细辛组血清中 IL-1β、TNF-α、NO 和 iNOS 含量均明显低于模型组。更为重要的是，二妙丸组和二妙丸加细辛组关节软骨中 BrdU、CD34 和 SDF-1 的表达明显高于模型组。与二妙丸组相比，二妙丸加细辛组对 OA 大鼠的治疗作用更强。