Effects of Tuina plus <italic style="font-style: italic">Chenyuangao</italic>（陈元膏）on the Apoptosis， Matrix Metalloproteinases and Serum Inflammatory Factors in Knee Osteoarthritis Model Rabbit with Yang Deficiency and Congealing Cold Syndrome

AIKEBAIER Gulaisaer; LIU Junchang; WANG Yifeng; YADI Maihereyi; HE Jing; LU Xusheng; ZHANG Xiaoting; GUO Lei

doi:10.13288/j.11-2166/r.2022.22.012

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Effects of Tuina plus Chenyuangao（陈元膏）on the Apoptosis， Matrix Metalloproteinases and Serum Inflammatory Factors in Knee Osteoarthritis Model Rabbit with Yang Deficiency and Congealing Cold Syndrome

更新时间：2022-11-11

- Effects of Tuina plus Chenyuangao（陈元膏）on the Apoptosis， Matrix Metalloproteinases and Serum Inflammatory Factors in Knee Osteoarthritis Model Rabbit with Yang Deficiency and Congealing Cold Syndrome
- Journal of Traditional Chinese Medicine Vol. 63, Issue 22, Pages: 2154-2161(2022)
- 作者机构：
  
  1.新疆医科大学，新疆维吾尔自治区乌鲁木齐市水磨沟区尚德北路567号，830017
  2.新疆医科大学附属中医医院
  3.新疆维吾尔自治区乌鲁木齐市第一人民医院分院
  4.新疆医科大学附属哈密市中心医院
  5.新疆维吾尔自治区伊犁州友谊医院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2022.22.012
  CLC：
- Received：22 April 2022，
  
  Revised：2022-06-28，
  
  Published：17 November 2022
- 稿件说明：
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古来撒尔·艾克拜尔 ,刘俊昌,王忆葑等.陈元膏摩法对阳虚寒凝型膝骨性关节炎模型兔关节软骨细胞凋亡、基质金属蛋白酶及血清炎性因子的影响[J].中医杂志,2022,63(22):2154-2161.

AIKEBAIER Gulaisaer,LIU Junchang,WANG Yifeng,et al.Effects of Tuina plus Chenyuangao（陈元膏）on the Apoptosis， Matrix Metalloproteinases and Serum Inflammatory Factors in Knee Osteoarthritis Model Rabbit with Yang Deficiency and Congealing Cold Syndrome[J].Journal of Traditional Chinese Medicine,2022,63(22):2154-2161.
古来撒尔·艾克拜尔 ,刘俊昌,王忆葑等.陈元膏摩法对阳虚寒凝型膝骨性关节炎模型兔关节软骨细胞凋亡、基质金属蛋白酶及血清炎性因子的影响[J].中医杂志,2022,63(22):2154-2161. DOI： 10.13288/j.11-2166/r.2022.22.012.

AIKEBAIER Gulaisaer,LIU Junchang,WANG Yifeng,et al.Effects of Tuina plus Chenyuangao（陈元膏）on the Apoptosis， Matrix Metalloproteinases and Serum Inflammatory Factors in Knee Osteoarthritis Model Rabbit with Yang Deficiency and Congealing Cold Syndrome[J].Journal of Traditional Chinese Medicine,2022,63(22):2154-2161. DOI： 10.13288/j.11-2166/r.2022.22.012.

摘要

目的

探讨陈元膏摩法治疗阳虚寒凝型膝骨性关节炎（KOA）的效用及可能作用机制。

方法

60只新西兰大白兔随机分为正常组、模型组、手法组、陈元膏组、陈元膏摩法组及扶他林组，每组10只。模型组和各干预组采用高分子绷带屈曲位固定+薄荷油、气候箱法建立兔阳虚寒凝型KOA模型。造模8周后，正常组和模型组不作干预；手法组给予单纯推拿手法；陈元膏组涂抹陈元膏剂；陈元膏摩法组在陈元膏组基础上行推拿手法治疗；扶他林组涂抹扶他林软膏，每日1次，连续6周。最后一次干预后次日进行取材，HE染色观察软骨、滑膜组织病理学变化并将切片进行病理学Mankin软骨组织学评分；ELISA法检测血清中炎性因子白细胞介素1β（IL-1β）、白细胞介素6（IL-6）、白细胞介素10（IL-10）、肿瘤坏死因子α（TNF-α）水平；Western Blot法检测B淋巴细胞瘤2（Bcl-2）、B淋巴细胞瘤2相关 X蛋白质（Bax）水平；免疫组化法及qRT-PCR法测定关节软骨组织中金属蛋白酶抑制剂1（TIMP-1）、基质金属蛋白酶13（MMP-13）、基质金属蛋白酶3（MMP-3）蛋白及mRNA表达水平。

结果

病理结果显示，正常组兔关节软骨细胞排列有序，基质均匀一致，滑膜组织结构正常，滑膜细胞排列整齐。模型组软骨基质失去均质性，部分区域软骨细胞崩解、消失、排列紊乱，滑膜细胞增大，层数增加，间质纤维组织增生伴部分玻璃样变性，较多慢性炎性细胞浸润。手法组、陈元膏组、陈元膏摩法组软骨细胞损伤均有不同程度改善，且陈元膏组及陈元膏摩法组改善程度优于手法组。与正常组比较，模型组兔关节软骨组织Mankin评分升高，血清IL-1β、TNF-α、IL-6水平升高，软骨组织Bcl-2蛋白表达及Bcl-2/Bax值降低，Bax、MMP-13蛋白表达升高，MMP-3、MMP-13 mRNA表达升高，TIMP-1 mRNA表达降低（

＜0.05）。与模型组比较，陈元膏摩法组兔关节软骨组织Mankin评分降低；各干预组血清IL-1β水平及软骨组织中MMP-3、MMP-13 mRNA水平降低，TIMP-1 mRNA水平升高；陈元膏摩法组、扶他林组IL-10水平、Bcl-2蛋白表达、Bcl-2/Bax值升高，Bax蛋白表达、MMP-13、TIMP-1蛋白表达降低（

＜0.05）。与扶他林组比较，陈元膏摩法组Mankin评分降低，Bax蛋白表达、MMP-13 mRNA表达升高（

＜0.05），两组各炎性因子水平差异无统计学意义（

＞0.05）。

结论

陈元膏摩法可改善软骨、滑膜病理组织形态，具有抗炎、抗软骨基质降解及抗软骨细胞凋亡作用，从而减轻关节软骨损伤，延缓阳虚寒凝型KOA病程进展。

Abstract

Objective

To observe the effect of tuina plus

Chenyuangao

（陈元膏） in the treatment of knee osteoarthritis （KOA） with yang deficiency and congealing cold syndrome， and to explore its potential mechanism.

Methods

Sixty New Zealand white rabbits were randomly divided into six groups， namely the normal group， model group， tuina group，

Chenyuangao

group， tuina plus

Chenyuangao

group and voltaren group， with ten rabbits per group. The model group and all intervention groups were treated by polymer bandage fixation at flexion angle combined with peppermint oil and climate box to establish rabbit model of KOA with yang deficiency and congealing cold syndrome. After eight weeks of modeling， the normal group and the model group were given no intervention. Tuina group was administered with tuina manipulation only， and

Chenyuangao

group was applied with ointment only， while tuina plus

Chenyuangao

group performed tuina based on the ointment

Chenyuangao

. The voltaren group was treated with voltaren ointment. Treatment was given once a day for 6 weeks. On the second day after the last intervention， samples were collected. Using HE staining to observe histopathological changes of cartilage and synovial membrane microscope， and the pathological Mankin score of the sections was assessed. Serum levels of inflammatory factors interleukin 1β （IL-1β）， interleukin 6 （IL-6）， interleukin-10 （IL-10） and tumor necrosis factor α （TNF-α） were detected by ELISA. Western Blot was used to detect the levels of B-lymphoma 2 （Bcl-2） and B-lymphoma 2-associated X protein （Bax）. The protein and mRNA expression levels of metalloproteinase inhibitor 1 （TIMP-1）， matrix metalloproteinase 13 （MMP-13） and matrix metalloproteinase 3 （MMP-3） in articular cartilage tissue were determined by immunohistochemistry and qRT-PCR.

Results

The pathological results showed that the articular chondrocytes of the rabbits in the normal group were arranged in an orderly manner， and the matrix was uniform， with normal synovial tissue structure and neatly arranged synovial cells. In the model group， the cartilage matrix lost its homogeneity， and the chondrocytes disintegrated， disappeared and disordered in certain areas， with increased synovial cells and number of layers， interstitial fibrous tissue proliferated with partial hyaline degeneration， and more chronic inflammatory cells infiltrated. The injury of cartilage cells in the tuina group， the

Chenyuangao

group and tuina plus

Chenyuangao

group were improved to varied degrees， and more changes were seen in the latter two groups. The model group had higher articular cartilage tissue Mankin score， higher levels of IL-1β， TNF-α， and IL-6， lower expression of Bcl-2 protein and Bcl-2/Bax value in the cartilage tissue， higher protein expressions of Bax and MMP-13， higher mRNA expressions of MMP-3 and MMP-13， and lower mRNA expression of TIMP-1 than the normal gorup （

0.05）. Compared to those in the model group， the Mankin score in the tuina plus

Chenyuangao

group significantly decreased； the serum IL-1β level and mRNA levels of MMP-3 and MMP-13 in cartilage tissue in other intervention groups significantly decreased， while the mRNA level of TIMP-1 increased； the IL-10 level， the expression of Bcl-2 protein and the Bcl-2/Bax value in the tuina plus

Chenyuangao

group and the voltaren group significantly increased， while the protein expression of Bax， MMP-13 and TIMP-1 decreased （

＜0.05）. Compared to those in the voltaren group， the Mankin score in the tuina plus

Chenyuangao

group was reduced， while the expressions of Bax protein and MMP-13 mRNA increased （

＜0.05）， and there was no significant difference between groups in the levels of inflammatory factors （

＞0.05）.

Conclusion

Tuina plus

Chenyuangao

can improve the histopathology of cartilage and synovial membrane， and has the functions of anti-inflammatory， anti-cartilage matrix degradation and anti-chondrocyte apoptosis， thereby reducing the damage of articular cartilage and delaying the progression of KOA with yang deficiency and congealing cold syndrome.

关键词

Keywords

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Effects of Tuina plus Chenyuangao（陈元膏）on the Apoptosis， Matrix Metalloproteinases and Serum Inflammatory Factors in Knee Osteoarthritis Model Rabbit with Yang Deficiency and Congealing Cold Syndrome

DOI：10.13288/j.11-2166/r.2022.22.012

摘要

Abstract

关键词

Keywords

references