基于lncRNA MALAT1调控软骨细胞胆固醇代谢探讨透骨消痛胶囊延缓骨关节炎退变的机制研究  

Mechanism of Tougu Xiaotong Capsules in delaying degeneration of osteoarthritis by regulating cholesterol metabolism in chondrocytes through lncRNA MALAT1

作　　者：付长龙[1] 林艳铭 涂海水 叶锦霞[1] 黄艳峰 马德尊 郑春松[1] FU Chang-long;LIN Yan-ming;TU Hai-shui;YE Jin-xia;HUANG Yan-feng;MA De-zun;ZHENG Chun-song(Fujian Key Laboratory of Integrative Medicine on Geriatrics,Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学、中西医结合研究院、福建省中西医结合老年性疾病重点实验室,福建福州350122

出　　处：《中国中药杂志》2024年第7期1785-1792,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82104888);福建中医药大学高层次人才科研启动资金项目(X2019011-人才);福建中医药大学校管课题(X2023024);陈可冀中西医结合发展基金项目(CKJ2021018,CKJ2023004)。

摘　　要：从长链非编码RNA肺腺癌转录本1(lncRNA MALAT1)调控软骨细胞胆固醇代谢角度,探讨透骨消痛胶囊(Tougu Xiaotong Capsules,TGXTC)延缓骨关节炎退变的作用与机制。8周龄C57BL/6小鼠48只经适应性喂养1周后,随机数字表法分为空白组12只、造模组36只,造模组动物经5%异氟醚吸入麻醉后,采用Hulth法诱导骨关节炎(OA)模型,随机分为模型组12只、阳性药组(牛磺熊去氧胆酸)12只、透骨消痛胶囊组12只。阳性药组予以牛磺熊去氧胆酸500 mg·kg^(-1)进行灌胃;透骨消痛胶囊组予以透骨消痛胶囊368 mg·kg^(-1)进行灌胃;空白组和模型组给予等量生理盐水灌服,连续灌胃4周。干预结束后,麻醉下处死各组小鼠,分离与收集膝关节软骨组织。通过形态学染色观察小鼠膝关节软骨组织结构变化;通过real-time PCR检测小鼠软骨组织中lncRNA MALAT1水平变化;通过Western blot检测小鼠关节软骨中ATP结合盒转运体A1(ABCA1)、载脂蛋白A1(ApoA1)、肝X核激素受体β(LXRβ)、C/EBP同源蛋白(CHOP)、胱天蛋白酶(caspase)-3蛋白表达情况。使用慢病毒包被的质粒进行敲减肺腺癌转录本1(sh-MALAT1)转染小鼠软骨细胞;real-time PCR检测sh-MALAT1转染小鼠软骨细胞中lncRNA MALAT1基因水平;Western blot检测透骨消痛胶囊对lncRNA MALAT1敲低后,毒胡萝卜素(thapsigargin,TG)诱导的小鼠软骨细胞ABCA1、ApoA1、LXRβ、CHOP、caspase-3蛋白含量表达情况。流式细胞术检测透骨消痛胶囊对lncRNA MALAT1敲低后TG诱导的小鼠软骨细胞凋亡率影响。苏木精-伊红(HE)染色与番红O染色结果显示,与模型组比较,透骨消痛胶囊组、阳性药组软骨层结构基本完整,关节结构破坏程度显著改善,软骨基质被番红O染色显著增强;与模型组比较,透骨消痛胶囊组、阳性药组中lncRNA MALAT1水平明显降低;与模型组比较,透骨消痛胶囊组、阳性药组的ABCA1、ApoA1、LXRβ蛋白含量表达显著增高,CHOP、caspase-3蛋白含From the perspective of lncRNA MALAT1 regulating cholesterol metabolism in chondrocytes,this paper explores the effect and mechanism of Tougu Xiaotong Capsules(TGXTC)in delaying the degeneration of osteoarthritis.After one week of adaptive feeding,48(8-week-old)C57BL/6 mice were randomly divided into a blank group(12 mice)and a model group(36 mice)by random number table method.The mice in the model group were anesthetized by inhalation of 5%isoflurane,and the OA model was induced by Hulth method.The experiment randomly divided the mice into a model group(12 mice),a drug-positive group(taurursodeoxycholic acid)(12 mice),and a TGXTC group(12 mice).The drug-positive group was given 500 mg·kg^(-1) taurodeoxycholic acid by intragastric administration.TGXTC group was given TGXTC 368 mg·kg^(-1) by gavage.The blank group and model group were given the same amount of normal saline for four weeks.After the intervention,the mice in each group were killed under anesthesia,and the knee cartilage tissue was separated and collected.The morphologic changes of knee cartilage were observed.The level of lncRNA MALAT1 in the cartilage tissue was detected by real-time PCR.The protein expressions of ABCA1,ApoA1,LXRβ,CHOP,and caspase-3 in mouse articular cartilage were detected by Western blot.Lentivirus-coated plasmid was used to transfect mouse chondrocytes with sh-MALAT1.The gene levels of lncRNA MALAT1 in mouse chondrocytes transfected with sh-MALAT1 were detected by real-time PCR.Western blot was used to detect the effect of TGXTC on the protein content of ABCA1,ApoA1,LXRβ,CHOP,and caspase-3 in thapsigargin(TG)-induced mouse chondrocytes after lncRNA MALAT1 knockdown.Flow cytometry was used to detect the effect of TGXTC on apoptosis of TG-induced mouse chondrocytes after lncRNA MALAT1 knockdown.The results of HE and saffranine O staining showed that compared with the model group,the structure of the cartilage layer was basically intact;the damage degree of joint structure was significantly improved,and the cartilage matrix was

关 键 词：透骨消痛胶囊 骨关节炎 胆固醇代谢 软骨退变 

分 类 号：R285.5[医药卫生—中药学]