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[1]陳俊,鄭若曦,葉錦霞,等.獨活寄生湯治療大鼠膝骨關(guān)節(jié)炎的作用機制研究[J].中醫(yī)正骨,2021,33(11):6-12.
　CHEN Jun,ZHENG Ruoxi,YE Jinxia,et al.Study on mechanism of action of Duhuo Jisheng Tang(獨活寄生湯)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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獨活寄生湯治療大鼠膝骨關(guān)節(jié)炎的作用機制研究()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第33卷
期數(shù):
2021年11期
頁碼:
6-12
欄目:
基礎(chǔ)研究
出版日期:
2021-11-20

文章信息/Info

Title:
Study on mechanism of action of Duhuo Jisheng Tang(獨活寄生湯)in treatment of knee osteoarthritis in rats
作者:
陳俊1; 鄭若曦2; 葉錦霞3; 鄭春松2; 張婷2; 戴雨婷2; 洪江從4; 李民4; 吳廣文2
(1.福建中醫(yī)藥大學(xué)中西醫(yī)結(jié)合學(xué)院,福建 福州 350122; 2.福建中醫(yī)藥大學(xué)中西醫(yī)結(jié)合研究院,福建 福州 350122; 3.福建省中西醫(yī)結(jié)合老年性疾病重點實驗室,福建 福州 350122; 4.福建中醫(yī)藥大學(xué)附屬康復(fù)醫(yī)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
關(guān)鍵詞:
骨關(guān)節(jié)炎; 膝;  軟骨組織;  獨活寄生湯;  大鼠; Sprague-Dawley;  RNA;  RNA; 長鏈非編碼;  聚集蛋白聚糖;  膠原Ⅱ型;  動物實驗
Keywords:
osteoarthritis; knee;  cartilaginous tissue;  Duhuo Jisheng Tang;  rats; Sprague-Dawley;  RNA;  RNA; long noncoding;  aggrecans;  collagen typeⅡ;  animal experimentation
摘要:
目的:探討?yīng)毣罴纳鷾委煷笫笙ス顷P(guān)節(jié)炎(knee osteoarthritis,KOA)的作用機制。方法:從66只大鼠中隨機選取48只,采用改良Hulth法建立大鼠KOA模型,隨機選擇3只正常大鼠和3只建模大鼠進行鑒定。證實建模成功后,將剩余的15只正常大鼠納入正常對照組,采用隨機數(shù)字表將剩余的45只KOA模型大鼠分為KOA模型組、獨活寄生湯治療組、鹽酸氨基葡萄糖治療組,每組15只。正常對照組、KOA模型組、獨活寄生湯治療組、鹽酸氨基葡萄糖治療組大鼠均按每天10 mL·kg-1分別以0.9%生理鹽水、0.9%生理鹽水、獨活寄生湯(濃度為0.93 g·mL-1)、鹽酸氨基葡萄糖溶液(濃度為0.015 g·mL-1)灌胃,連續(xù)干預(yù)12周。12周后,切取大鼠雙側(cè)脛骨近端和股骨下端的軟骨組織。采用實時定量PCR檢測大鼠脛骨近端軟骨組織中miR-675-3p、長鏈非編碼RNA(long non-coding RNA,lncRNA)H19及聚集蛋白聚糖(Aggrecan)mRNA、Ⅱ型膠原蛋白(CollagenⅡ)mRNA的表達量,采用蛋白印跡法檢測大鼠股骨遠端軟骨組織中Aggrecan、CollagenⅡ的蛋白表達量。結(jié)果:①大鼠飼養(yǎng)結(jié)果。KOA模型組大鼠因肺栓塞死亡2只,獨活寄生湯治療組大鼠因腹瀉死亡1只,鹽酸氨基葡萄糖治療組大鼠因肺栓塞死亡1只。②大鼠脛骨近端軟骨組織中KOA相關(guān)基因的RNA表達結(jié)果。正常對照組、KOA模型組、獨活寄生湯治療組、鹽酸氨基葡萄糖治療組大鼠脛骨近端軟骨組織中miR-675-3p、lncRNA H19、Aggrecan mRNA、CollagenⅡmRNA的表達量比較,組間差異均有統(tǒng)計學(xué)意義(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模型組、獨活寄生湯治療組、鹽酸氨基葡萄糖治療組大鼠脛骨近端軟骨組織中miR-675-3p、Aggrecan mRNA、CollagenⅡmRNA的表達量均低于正常對照組(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模型組大鼠脛骨近端軟骨組織中l(wèi)ncRNA H19的表達量低于正常對照組(P=0.002),獨活寄生湯治療組、鹽酸氨基葡萄糖治療組大鼠脛骨近端軟骨組織中l(wèi)ncRNA H19的表達量與正常對照組比較,差異無統(tǒng)計學(xué)意義(P=0.058,P=0.069); 獨活寄生湯治療組、鹽酸氨基葡萄糖治療組大鼠脛骨近端軟骨組織中miR-675-3p、Aggrecan mRNA、CollagenⅡmRNA的表達量均高于KOA模型組(miR-675-3p:P=0.000; P=0.000; Aggrecan mRNA:P=0.000; P=0.000; CollagenⅡmRNA:P=0.000; P=0.000); 獨活寄生湯治療組大鼠脛骨近端軟骨組織中miR-675-3p、lncRNA H19、Aggrecan mRNA、CollagenⅡmRNA的表達量與鹽酸氨基葡萄糖治療組比較,組間差異均無統(tǒng)計學(xué)意義(P=0.922; P=0.778; P=0.857; P=0.904)。③大鼠股骨遠端軟骨組織中軟骨組成相關(guān)基因的蛋白表達結(jié)果。正常對照組、KOA模型組、獨活寄生湯治療組、鹽酸氨基葡萄糖治療組的大鼠股骨遠端軟骨組織中Aggrecan和CollagenⅡ的蛋白表達量比較,組間差異均有統(tǒng)計學(xué)意義(F=58.118,P=0.002; F=16.625,P=0.001)。KOA模型組、獨活寄生湯治療組大鼠股骨遠端軟骨組織中Aggrecan的蛋白表達量均低于正常對照組(P=0.003; P=0.004),鹽酸氨基葡萄糖治療組大鼠股骨遠端軟骨組織中Aggrecan的蛋白表達量與正常對照組相比,差異無統(tǒng)計學(xué)意義(P=0.078),KOA模型組、獨活寄生湯治療組、鹽酸氨基葡萄糖治療組大鼠股骨遠端軟骨組織中CollagenⅡ的蛋白表達量均低于正常對照組(P=0.000; P=0.028; P=0.018),獨活寄生湯治療組、鹽酸氨基葡萄糖治療組大鼠股骨遠端軟骨組織中Aggrecan和CollagenⅡ的蛋白表達量均高于KOA模型組(Aggrecan:P=0.025; P=0.034; CollagenⅡ:P=0.003; P=0.004),獨活寄生湯治療組大鼠股骨遠端軟骨組織中Aggrecan和CollagenⅡ的蛋白表達量與鹽酸氨基葡萄糖治療組比較,組間差異均無統(tǒng)計學(xué)意義(P=0.999; P=0.775)。結(jié)論:獨活寄生湯治療大鼠KOA的作用機制,可能與其能夠上調(diào)miR-675-3p、lncRNA H19的表達進而促進Aggrecan、CollagenⅡ的表達有關(guān)。
Abstract:
Objective:To explore the mechanism of action(MoA)of Duhuo Jisheng Tang(獨活寄生湯,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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備注/Memo

備注/Memo:
基金項目:國家自然科學(xué)基金項目(81774345)
通訊作者:吳廣文 E-mail:[email protected]
更新日期/Last Update: 1900-01-01