摘要
目的 基于半胱氨酸蛋白酶3/多聚腺苷二磷酸核糖聚合酶(Caspase-3/PARP)通路探究龟鹿二仙胶及拆方对白细胞介素-1β(IL-1β)诱导的退变软骨细胞凋亡及细胞外基质(ECM)的影响。方法 采用SD大鼠制备空白血清、龟鹿二仙胶含药血清、龟甲胶含药血清及鹿角胶含药血清,采用酶消法体外培养C57BL/6小鼠软骨细胞。将软骨细胞分为5组,空白组加入空白血清培养,模型组加入IL-1β和空白血清培养,龟鹿组加入IL-1β和龟鹿二仙胶含药血清培养,龟板组加入IL-1β和龟甲胶含药血清培养,鹿角组加入IL-1β和鹿角胶含药血清培养,干预24 h后,采用CCK-8法检测软骨细胞活性,采用TUNEL法检测软骨细胞凋亡情况,采用免疫荧光染色法检测Caspase-3、PARP-1表达情况,分别采用Western blot和qRT-PCR法检测细胞中Caspase-3、PARP-1、基质金属蛋白酶-13(MMP-13)、聚集蛋白聚糖(Aggrecan)蛋白及mRNA表达情况。结果 与空白组比较,模型组软骨细胞活性明显降低(P<0.05),软骨细胞凋亡率明显升高(P<0.05);细胞中Caspase-3、PARP-1平均荧光强度和Caspase-3、PARP-1、MMP-13蛋白及mRNA相对表达量均明显升高(P均<0.05),细胞中Aggrecan蛋白及mRNA相对表达量均明显降低(P均<0.05)。与模型组比较,龟鹿组、龟板组及鹿角组软骨细胞活性均明显升高(P均<0.05),软骨细胞凋亡率均明显降低(P均<0.05);细胞中Caspase-3、PARP-1平均荧光强度和Caspase-3、PARP-1、MMP-13蛋白及mRNA相对表达量均明显降低(P均<0.05),细胞中Aggrecan蛋白及mRNA相对表达量均明显升高(P均<0.05)。与龟板组、鹿角组比较,龟鹿组软骨细胞活性更高(P均<0.05),软骨细胞凋亡率更低(P均<0.05);细胞中Caspase-3、PARP-1平均荧光强度和Caspase-3、PARP-1、MMP-13蛋白及mRNA相对表达量均更低(P均<0.05),细胞中Aggrecan蛋白及mRNA相对表达量均更高(P均<0.05)。与龟板组比较,鹿角组软骨细胞活性,软骨细胞凋亡率,细胞中Caspase-
Objective It is to investigate the effects of decoction of Tortoise Plastron Glue plus Deerhorn Glue and its decomposed on interleukin-1β(IL-1β)induced apoptosis and extracellular matrix(ECM)of chondrocytes via cysteine aspartic protease 3(Caspase-3)/poly ADP-ribose polymerase(PARP)pathway.Methods The blank serum,decoction of Tortoise Plastron Glue plus Deerhorn Glue containing serum,Tortoise Plastron Glue containing serum and Deerhorn Glue containing serum were prepared using SD rats,and the C57BL/6 mouse chondrocytes were cultured by enzyme ablation method in vitro.The chondrocytes were divided into 5 groups:blank group cultured with blank containing serum,model group cultured with IL-1βand blank containing serum,Tortoise Plastron Glue plus Deerhorn Glue group cultured with IL-1βand Tortoise Plastron Glue plus Deerhorn Glue containing serum,Tortoise Plastron Glue group cultured with IL-1βand Tortoise Plastron Glue containing serum,Deerhorn Glue group cultured with IL-1βand Deerhorn Glue containing serum.After 24h of intervention,the activity of chondrocyte was detected by CCK-8 method and its apoptosis was detected by TUNEL method,the expressions of PARP-1 and Caspase-3 were detected by immunofluorescence staining,the protein and mRNA expressions of PARP-1,Caspase-3,matrix Metallopeptidase 13(MMP-13),aggrecan were detected by Western blotting and qRT-PCR.Results Compared with the blank group,the activity of chondrocytes in the model group was significantly decreased(P<0.05),and the apoptosis rate of chondrocytes was significantly increased(P<0.05);the average fluorescence intensity of PARP-1 and Caspase-3,the protein and mRNA expressions of PARP-1,Caspase-3,MMP-13 in the cells were significantly increased(all P<0.05),while the protein and mRNA expressions of Aggrecan were significantly decreased(all P<0.05).Compared with the model group,the activities of chondrocytes in the Tortoise Plastron Glue plus Deerhorn Glue group,Tortoise Plastron Glue group,Deerhorn Glue group were significantly increased(all P<0
作者
吴伟欣
郑珍萍
顾富城
杨美鑫
王和鸣
李楠
WU Weixin;ZHENG Zhenping;GU Fucheng;YANG Meixin;WANG Heming;LI Nan(College of Traditional Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian,China;Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian,China;Key Laboratory of Traditional Chinese Medicine for Bone Injury and Sports Rehabilitation,Ministry of Education,Fuzhou 350122,Fujian,China)
出处
《现代中西医结合杂志》
CAS
2024年第1期8-16,共9页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
国家自然科学基金面上项目(81973880)
福建省自然科学基金项目(2022J01364)。
关键词
龟鹿二仙胶
软骨细胞
凋亡
细胞外基质
半胱氨酸蛋白酶3
多聚腺苷二磷酸核糖聚合酶
decoction of Tortoise Plastron Glue plus Deerhorn Glue
chondrocyte
apoptosis
extracellular matrix
cysteine aspartic protease 3
poly ADP-ribose polymerase
