右归丸对大鼠膝骨关节炎的干预作用及其机制*

doi:10.12047/j.cjap.5988.2020.109

中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (5): 511-516.doi: 10.12047/j.cjap.5988.2020.109

右归丸对大鼠膝骨关节炎的干预作用及其机制^*

颜春鲁^1,7, 安方玉^1,6△, 汪永锋³, 金华⁴, 赵崇博⁵, 王统香⁵, 邓婕⁵, 张蕾蕾⁸

1.甘肃中医药大学中西医结合学院, 2.教学实验实训中心, 3.基础医学院, 4.中医临床学院, 5.临床医学院, 甘肃兰州 730000;
6.甘肃省高校重大疾病分子医学与中医药防治研究重点实验室, 甘肃兰州 730000;
7.敦煌医学与转化省部共建教育部重点实验室, 甘肃兰州 730000;
8.兰州市疾病预防控制中心, 甘肃兰州 730000

收稿日期:2019-12-16 修回日期:2020-06-18 发布日期:2021-02-25
通讯作者: ^△Tel: 13893639394; E-mail: anfyuwsmh@163.com
基金资助:
^*甘肃省科技厅自然科学基金(20JR5RA185);甘肃省高等学校创新能力提升项目(2019A-080);甘肃省高校大学生就业创业能力提升工程项目(6-1);甘肃省高校中(藏)药化学与质量研究省级重点实验室开放基金项目(zzy-2018-01);甘肃省高校重大疾病分子医学与中医药防治研究重点实验室开放基金项目(FZYX17-18-11)

Intervention effect of Youguiwan on rats with knee osteoarthritis and its mechanism

YAN Chun-lu^1,7, AN Fang-yu^1,6△, WANG Yong-feng³, JIN Hua⁴, ZHAO Chong-bo⁵, WANG Tong-xiang⁵, DENG Jie⁵, ZHANG Lei-lei⁸

1. Institute of Chinese and Western Medicine, Gansu University of Traditional Chinese Medicine, Lanzhou 730000;2. Teaching Experiment Training Center;3. School of Basic Medical Science;4. Clinical College of Traditional Chinese Medicine;5. School of Clinical Medicine;
6. Provincial-Level Key Laboratory for Molecular Medicine of Major Diseases and the Study on Prevention and Treatment of Traditional Chinese Medicine, Gansu University of Traditional Chinese Medicine, Lanzhou 730000;
7. Key Laboratory of Dunhuang Medicine, Ministry of Education, Lanzhou 730000;
8. Centers for Disease Control and Prevention of Lanzhou, Lanzhou 730000, China

Received:2019-12-16 Revised:2020-06-18 Published:2021-02-25

摘要/Abstract

摘要： 目的: 探讨右归丸对膝骨关节炎(KOA)模型鼠关节软骨组织骨诱导因子(OGN)、骨黏连蛋白(ON)和纤维蛋白原2(FBN2)的影响。方法: 将大鼠随机分为假手术组,模型组,硫酸氨基葡萄糖组(硫酸氨基葡萄糖),右归丸(高、中、低剂量)组,每组10只。采用改良Hulth法制备大鼠KOA模型,假手术组和模型组给予等体积生理盐水灌胃,右归丸高、中、低剂量组分别灌胃给予右归丸4.8,2.4,1.2 g/kg,硫酸氨基葡萄糖组灌胃给予硫酸氨基葡萄糖 0.17 g/kg,连续给药8周。干预结束24 h后取鼠膝关节软骨,采用HE染色法观察各组软骨的病理改变,并进行Mankin评分;免疫组化法检测各组关节软骨组织中OGN、ON和FBN2的表达;Western blot法检测各组关节软骨组中糖原合成酶激酶-3β(GSK-3β)的表达。结果: 与假手术组比较,模型组大鼠软骨组织Makin 评分显著升高,软骨组织FBN2蛋白表达水平上显著增加,OGN、ON和GSK-3β蛋白表达水平上的显著降低(P＜0.01);模型组关节软骨边缘严重破坏,软骨细胞排列紊乱。与模型组比较,右归丸高剂量干预组大鼠软骨组织Makin 评分和FBN2蛋白表达水平显著降低,GSK-3β蛋白表达水平上显著增加,且右归丸中、高剂量组OGN、ON蛋白表达水平均显著增加(P＜0.05或P＜0.01),软骨结构趋于正常,软骨细胞分布仅偶见不均,关节软骨表面欠光滑。结论: 右归丸能够延缓关节软骨退变,其可能机制是通过提高骨诱导因子和骨粘连蛋白的表达水平来促进关节软骨的骨化和重构。

关键词: 膝骨关节炎, 右归丸, 骨诱导因子, 骨黏连蛋白, 纤维蛋白原2

Abstract: Objective: To study the effects of Youguiwan on the osteoglycin (OGN), osteonectin (ON), fibrinogen 2 (FBN2) of articular cartilage tissue in the model of knee osteoarthritis (KOA). Methods: Sixty SD rats were randomly divided into six groups: sham control group, model group, glucosamine sulfate group, Youguiwan (high-dose, middle-dose and low-dose )group. The modified Hulth method was used to establish KOA models for 6 weeks. The sham control group and the model group were treated with normal saline. The rats in Youguiwan high-dose, middle-dose, low-dose groups were treated with Youguiwan at the doses of 4.8, 2.4, 1.2 g/kg by gavage respectively, and the glucosamine sulfate group was treated with glucosamine sulfate 0.17 g/kg. The rats were administrated for 8 weeks according to the dose. After intervening, articular cartilage of rats were obtained, the pathological changes were observed by using HE staining method, and Mankin score was evaluated. The expressions of OGN, ON and FBN2 in articular cartilage were detected by immunohistochemistry. The expression of GSK-3β in articular cartilage was detected by Western blot. Results: Compared with the sham control group, the Mankin score was obviously increased in the model group, the protein expression of FBN2 was increased significantly, yet the protein expressions of OGN, ON and GSK-3β were decreased significantly (P＜0.01), articular cartilage was seriously damaged, and chondrocytes were arranged in disorder. Compared with the model group, the Mankin score was declined obviously in the high-dose Youguiwan group, the protein expression of FBN2 was significantly decreased, but the protein expression of GSK-3β was significantly increased, the protein expressions of OGN and ON were significantly increased in the middle-dose and high-dose Youguiwan group (P＜0.05 or P＜0.01), cartilage structure was tended to be normal, the chondrocytes distribution was uneven, and articular cartilage surface was not smooth. Conclusion: Youguiwan can significantly improve the articular cartilage degeneration of KOA rats, its mechanism maybe raise OGN and ON protein expression level to promote the ossification and reconstruction of articular cartilage.

Key words: knee osteoarthritis, youguiwan, osteoglycin, osteonectin, fibrinogen 2

中图分类号:

R684.3

颜春鲁, 安方玉, 汪永锋, 金华, 赵崇博, 王统香, 邓婕, 张蕾蕾. 右归丸对大鼠膝骨关节炎的干预作用及其机制^*[J]. 中国应用生理学杂志, 2020, 36(5): 511-516.

YAN Chun-lu, AN Fang-yu, WANG Yong-feng, JIN Hua, ZHAO Chong-bo, WANG Tong-xiang, DENG Jie, ZHANG Lei-lei. Intervention effect of Youguiwan on rats with knee osteoarthritis and its mechanism[J]. CJAP, 2020, 36(5): 511-516.

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右归丸对大鼠膝骨关节炎的干预作用及其机制^*

Intervention effect of Youguiwan on rats with knee osteoarthritis and its mechanism

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