[1]陈俊,郑若曦,叶锦霞,等.独活寄生汤治疗大鼠膝骨关节炎的作用机制研究[J].中医正骨,2021,33(11):6-12.
　CHEN Jun,ZHENG Ruoxi,YE Jinxia,et al.Study on mechanism of action of Duhuo Jisheng Tang(独活寄生汤)in treatment of knee osteoarthritis in rats[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2021,33(11):6-12.
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独活寄生汤治疗大鼠膝骨关节炎的作用机制研究()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第33卷
期数:
2021年11期
页码:
6-12
栏目:
基础研究
出版日期:
2021-11-20

文章信息/Info

Title:
Study on mechanism of action of Duhuo Jisheng Tang(独活寄生汤)in treatment of knee osteoarthritis in rats
作者:
陈俊1; 郑若曦2; 叶锦霞3; 郑春松2; 张婷2; 戴雨婷2; 洪江从4; 李民4; 吴广文2
(1.福建中医药大学中西医结合学院,福建 福州 350122; 2.福建中医药大学中西医结合研究院,福建 福州 350122; 3.福建省中西医结合老年性疾病重点实验室,福建 福州 350122; 4.福建中医药大学附属康复医院,福建 福州 350003)
Author(s):
CHEN Jun1; ZHENG Ruoxi2; YE Jinxia3; ZHENG Chunsong2; ZHANG Ting2; DAI Yuting2; HONG Jiangcong4; LI Min4; WU Guangwen2
1.College of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian,China2.Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian,China3.Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350122,Fujian,China4.Rehabilitation Hospital affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou 350003,Fujian,China
关键词:
骨关节炎; 膝;  软骨组织;  独活寄生汤;  大鼠; Sprague-Dawley;  RNA;  RNA; 长链非编码;  聚集蛋白聚糖;  胶原Ⅱ型;  动物实验
Keywords:
osteoarthritis; knee;  cartilaginous tissue;  Duhuo Jisheng Tang;  rats; Sprague-Dawley;  RNA;  RNA; long noncoding;  aggrecans;  collagen typeⅡ;  animal experimentation
摘要:
目的:探讨独活寄生汤治疗大鼠膝骨关节炎(knee osteoarthritis,KOA)的作用机制。方法:从66只大鼠中随机选取48只,采用改良Hulth法建立大鼠KOA模型,随机选择3只正常大鼠和3只建模大鼠进行鉴定。证实建模成功后,将剩余的15只正常大鼠纳入正常对照组,采用随机数字表将剩余的45只KOA模型大鼠分为KOA模型组、独活寄生汤治疗组、盐酸氨基葡萄糖治疗组,每组15只。正常对照组、KOA模型组、独活寄生汤治疗组、盐酸氨基葡萄糖治疗组大鼠均按每天10 mL·kg-1分别以0.9%生理盐水、0.9%生理盐水、独活寄生汤(浓度为0.93 g·mL-1)、盐酸氨基葡萄糖溶液(浓度为0.015 g·mL-1)灌胃,连续干预12周。12周后,切取大鼠双侧胫骨近端和股骨下端的软骨组织。采用实时定量PCR检测大鼠胫骨近端软骨组织中miR-675-3p、长链非编码RNA(long non-coding RNA,lncRNA)H19及聚集蛋白聚糖(Aggrecan)mRNA、Ⅱ型胶原蛋白(CollagenⅡ)mRNA的表达量,采用蛋白印迹法检测大鼠股骨远端软骨组织中Aggrecan、CollagenⅡ的蛋白表达量。结果:①大鼠饲养结果。KOA模型组大鼠因肺栓塞死亡2只,独活寄生汤治疗组大鼠因腹泻死亡1只,盐酸氨基葡萄糖治疗组大鼠因肺栓塞死亡1只。②大鼠胫骨近端软骨组织中KOA相关基因的RNA表达结果。正常对照组、KOA模型组、独活寄生汤治疗组、盐酸氨基葡萄糖治疗组大鼠胫骨近端软骨组织中miR-675-3p、lncRNA H19、Aggrecan mRNA、CollagenⅡmRNA的表达量比较,组间差异均有统计学意义(F=83.050,P=0.000; F=436.003,P=0.000; F=129.212,P=0.000; F=135.844,P=0.000)。KOA模型组、独活寄生汤治疗组、盐酸氨基葡萄糖治疗组大鼠胫骨近端软骨组织中miR-675-3p、Aggrecan mRNA、CollagenⅡmRNA的表达量均低于正常对照组(miR-675-3p:P=0.000; P=0.003; P=0.002; Aggrecan mRNA:P=0.000; P=0.000; P=0.000; CollagenⅡmRNA:P=0.000; P=0.000; P=0.000); KOA模型组大鼠胫骨近端软骨组织中lncRNA H19的表达量低于正常对照组(P=0.002),独活寄生汤治疗组、盐酸氨基葡萄糖治疗组大鼠胫骨近端软骨组织中lncRNA H19的表达量与正常对照组比较,差异无统计学意义(P=0.058,P=0.069); 独活寄生汤治疗组、盐酸氨基葡萄糖治疗组大鼠胫骨近端软骨组织中miR-675-3p、Aggrecan mRNA、CollagenⅡmRNA的表达量均高于KOA模型组(miR-675-3p:P=0.000; P=0.000; Aggrecan mRNA:P=0.000; P=0.000; CollagenⅡmRNA:P=0.000; P=0.000); 独活寄生汤治疗组大鼠胫骨近端软骨组织中miR-675-3p、lncRNA H19、Aggrecan mRNA、CollagenⅡmRNA的表达量与盐酸氨基葡萄糖治疗组比较,组间差异均无统计学意义(P=0.922; P=0.778; P=0.857; P=0.904)。③大鼠股骨远端软骨组织中软骨组成相关基因的蛋白表达结果。正常对照组、KOA模型组、独活寄生汤治疗组、盐酸氨基葡萄糖治疗组的大鼠股骨远端软骨组织中Aggrecan和CollagenⅡ的蛋白表达量比较,组间差异均有统计学意义(F=58.118,P=0.002; F=16.625,P=0.001)。KOA模型组、独活寄生汤治疗组大鼠股骨远端软骨组织中Aggrecan的蛋白表达量均低于正常对照组(P=0.003; P=0.004),盐酸氨基葡萄糖治疗组大鼠股骨远端软骨组织中Aggrecan的蛋白表达量与正常对照组相比,差异无统计学意义(P=0.078),KOA模型组、独活寄生汤治疗组、盐酸氨基葡萄糖治疗组大鼠股骨远端软骨组织中CollagenⅡ的蛋白表达量均低于正常对照组(P=0.000; P=0.028; P=0.018),独活寄生汤治疗组、盐酸氨基葡萄糖治疗组大鼠股骨远端软骨组织中Aggrecan和CollagenⅡ的蛋白表达量均高于KOA模型组(Aggrecan:P=0.025; P=0.034; CollagenⅡ:P=0.003; P=0.004),独活寄生汤治疗组大鼠股骨远端软骨组织中Aggrecan和CollagenⅡ的蛋白表达量与盐酸氨基葡萄糖治疗组比较,组间差异均无统计学意义(P=0.999; P=0.775)。结论:独活寄生汤治疗大鼠KOA的作用机制,可能与其能够上调miR-675-3p、lncRNA H19的表达进而促进Aggrecan、CollagenⅡ的表达有关。
Abstract:
Objective:To explore the mechanism of action(MoA)of Duhuo Jisheng Tang(独活寄生汤,DHJST)in treatment of knee osteoarthritis(KOA)in rats.Methods:Among the 66 rats,48 ones were selected by using random digits table and subjected to surgery for inducing KOA by using modified Hulth method.Three normal rats and 3 model rats were randomly selected for confirming whether the models were built successfully.After successful modeling,the remained 15 normal rats were assigned into the normal control group,and the 45 KOA model rats were randomly divided into KOA model group,DHJST treatment group and glucosamine hydrochloride(GAH)treatment group by using random digits table,15 cases in each group.The rats in normal control group,KOA model group,DHJST treatment group and GAH treatment group were intragastric administrated with 0.9% normal saline(NS),0.9% NS,DHJST with concentration of 0.93 g/mL and GAH with concentration of 0.015 g/mL in daily dosage of 10 mL/kg respectively for consecutive 12 weeks.After the end of intervention,the cartilage tissues were harvested from bilateral proximal tibiae and terminal femora of rats in each group.The expression levels of miR-675-3p,long non-coding RNA(lncRNA)H19,Aggrecan mRNA and CollagenⅡmRNA in cartilage tissues of proximal tibiae were detected by using real-time quantitative PCR(RT-qPCR),and the protein expression levels of Aggrecan and CollagenⅡin cartilage tissues of terminal femora were detected by using Western-blot assays.Results:①Two rats in KOA model group,1 case in DHJST treatment group and 1 case in GAH treatment group died for pulmonary embolism(PE),diarrhea and PE respectively.②The relults of analysis on RNA expression of KOA-related genes in cartilage tissues of rat proximal tibiae.There was statistical difference in the expression levels of miR-675-3p,lncRNA H19,Aggrecan mRNA and CollagenⅡmRNA in cartilage tissues of rat proximal tibiae among normal control group,KOA model group,DHJST treatment group and GAH treatment group(F=83.050,P=0.000; F=436.003,P=0.000; F=129.212,P=0.000; F=135.844,P=0.000).The miR-675-3p,Aggrecan mRNA and CollagenⅡmRNA were lowly expressed in cartilage tissues of rat proximal tibiae in KOA model group,DHJST treatment group and GAH treatment group compared to normal control group(miR-675-3p:P=0.000; P=0.003; P=0.002; P=0.000; Aggrecan mRNA:P=0.000; P=0.000; P=0.000; CollagenⅡmRNA:P=0.000; P=0.000; P=0.000)...

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金项目(81774345)
通讯作者:吴广文 E-mail:wuguangwenfj@163.com
更新日期/Last Update: 1900-01-01