[1]张锐,马继海,柴喜平,等.独活寄生汤对膝骨关节炎模型大鼠NF-κB通路关键分子表达的影响[J].西部中医药,2023,36(08):15-19.[doi:10.12174/j.issn.2096-9600.2023.08.04]
　ZHANG Rui,MA Jihai,CHAI Xiping,et al.Influence of Duhuo Jisheng Decoction on the Expression of Key Molecules of NF-κB Pathway in Knee Osteoarthritis Rat Models[J].Western Journal of Traditional Chinese Medicine,2023,36(08):15-19.[doi:10.12174/j.issn.2096-9600.2023.08.04]
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独活寄生汤对膝骨关节炎模型大鼠NF-κB通路关键分子表达的影响
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《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

卷:
36
期数:
2023年08期
页码:
15-19
栏目:
出版日期:
2023-08-15

文章信息/Info

Title:
Influence of Duhuo Jisheng Decoction on the Expression of Key Molecules of NF-κB Pathway in Knee Osteoarthritis Rat Models
作者:
张锐, 马继海, 柴喜平, 李爱强, 柳海平
甘肃省中医院，甘肃 兰州 730050
Author(s):
ZHANG Rui, MA Jihai, CHAI Xiping, LI Aiqiang, LIU Haiping
Gansu Provincial Hospital of Traditional Chinese Medicine, Lanzhou 730050, China
关键词:
骨关节炎; 膝; 独活寄生汤; NF-B通路; 大鼠; 动物实验
Keywords:
osteoarthritis;  knee; decoction; NF-B pathway; rat; zoopery
分类号:
R285.5
DOI:
10.12174/j.issn.2096-9600.2023.08.04
文献标志码:
A
摘要:
目的探讨独活寄生汤对膝骨关节炎模型大鼠核因子κB（nuclear factor-kappaB，NF-κB）通路关键分子表达的影响。 方法将60只SPF级Wistar大鼠分为空白组（10只）和造模组（50只）。模型制备成功后按随机数字表法分为模型组，硫酸氨基葡萄糖组（阳性组），独活寄生汤高、中、低剂量组，共5组，每组10只。空白组及模型组大鼠给予生理盐水灌胃，阳性组及独活寄生汤各剂量组给予等体积药物灌胃，连续灌胃21天。观察各组大鼠一般生存状况，苏木精-伊红染色法观察各组大鼠软骨组织病理变化，酶联免疫吸附法检测大鼠血清白细胞介素4（interleukin-4，IL-4）、白细胞介素6（interleukin-6，IL-6）、白细胞介素10（interleukin-10，IL-10）及肿瘤坏死因子α（tumor necrosis factor α，TNF-α）的含量，荧光定量PCR（quantitative polymerase chain reaction，qPCR）检测各组软骨组织IL-4、IL-6、IL-10、TNF-α及NF-κB p65的mRNA表达水平，蛋白质印迹法检测各组软骨组织NF-κB p65及p-NF-κB p65的蛋白表达水平。 结果模型组大鼠精神萎靡，毛发凌乱，饮食饮水较少；独活寄生汤各剂量组和阳性组大鼠精神状态较好，毛发饮食等恢复正常，其中高剂量和阳性组恢复最好。空白组大鼠软骨组织细胞排列整齐，结构清晰，软骨基质着色均匀；模型组大鼠细胞排列紊乱，结构不清，表面粗糙；与模型组比较，阳性组和独活寄生汤各剂量组大鼠软骨组织结构较完整、清晰，表面较光滑，软骨基质着色较均匀。与空白组比较，模型组IL-4、IL-6及TNF-α血清中含量及软骨组织mRNA表达水平均升高，IL-10水平降低（P＜0.05）；与空白组比较，模型组软骨组织中NF-κB p65 mRNA及蛋白水平均升高（P＜0.05）；与模型组比较，独活寄生汤各剂量组、阳性组的血清的IL-4、IL-6、TNF-α含量及NF-κB p65、p-NF-κB p65蛋白水平均降低（P＜0.05）；与模型组比较，独活寄生汤中、高剂量组、阳性组IL-4、IL-6、TNF-α及NF-κB p65软骨组织mRNA水平均降低（P＜0.05）；血清IL-10含量、软骨组织mRNA水平升高（P＜0.05）。 结论独活寄生汤可调控IL-4、IL-6、IL-10、TNF-α、NF-κB p65及p-NF-κB p65表达，可能是治疗膝骨关节炎的机制之一。
Abstract:
ObjectiveTo discuss the effects of Duhuo Jisheng decoction on the expressions of key molecules of NF-κB pathway in rat models with knee osteoarthritis (KOA). MethodsSixty SPF grade Wistar rats were allocated to the blank group (10 rats) and the modeling group (50 rats). After successful modeling, the modeling group were divided into the model group, glucosamine sulfate group (positive group), high, moderate and low doses groups of Duhuo Jisheng decoction in light of random number table method, five groups in all, ten in each group. The blank group and the model group were drenched with physiological saline, positive group and different doses groups of Duhuo Jisheng decoction with equal volume of drugs, for 21 days consecutively by gavage. To observe general living situation of the rats in different groups, hematoxylin-eosin staining method was used to observe histopathological changes of the cartilage of the rats in different groups, ELISA to detect the contents of IL-4, IL-6 and IL-10, TNF-α, qPCR to measure the expressions of IL-4, IL-6 and IL-10, TNF-α and NF-κB p65 mRNA in cartilage in different groups, Western blotting method to test the levels of NF-κB p65 and p-NF-κB p65 protein in cartilage in different groups. ResultsThe rats in the model group were depressed, with messy hair and less food and water; the rats in high, moderate and low doses groups of Duhuo Jisheng decoction were in good mental condition, and their hair and diet returned to normal, among them, high dose group and positive group recovering at the best. Cartilage tissue cells of the rats in the blank group were neatly arranged and clearly structured, and the cartilage matrix is uniformly colored; the rats in the model group has disordered cell arrangement, poor structure and rough surface; compared with the model group, cartilage tissue were relatively complete and clear, with smooth surface, and the cartilage matrix was colored more evenly in positive group, high, moderate and low doses groups of Duhuo Jisheng decoction. Compared with the blank group, the contents of IL-4, IL-6 and TNF-α in the serum and the expression of mRNA in cartilage increased in the model group, the level of IL-10 decreased (P<0.05); compared with the blank group, the levels of NF-κB p65 mRNA and protein lifted in the model group (P<0.05); compared with the model group, the contents of serum IL-4, IL-6 and TNF-α, the levels of NF-κB p65 and p-NF-κB p65 lowered in different doses groups of Duhuo Jisheng decoction and positive group (P<0.05); compared with the model group, the levels of IL-4, IL-6 and TNF-α, NF-κB p65 cartilage mRNA in moderate and high dose groups of Duhuo Jisheng decoction, positive group reduced (P<0.05); the contents of serum IL-10 and cartilage mRNA level lifted (P<0.05). ConclusionDuhuo Jisheng decoction could regulate the expressions of IL-4, IL-6 and IL-10, TNF-α, NF-κB p65 and p-NF-κB p65, which may be one of the mechanisms of treating KOA.

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备注/Memo

备注/Memo:
张锐（1982—），男，硕士学位，副主任医师。研究方向：骨关节疾病的诊治。甘肃省自然科学基金（21JR1RA059）。
更新日期/Last Update: 2023-08-15