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基于IHH-Gli信号通路探讨龟鹿二仙胶含药血清调控大鼠膝关节软骨细胞肥大分化作用机制
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作者 李晔 耿秋东 +2 位作者 郑珍萍 吴伟欣 李楠 《福建中医药》 2023年第2期26-30,共5页
目的基于IHH-Gli信号通路探讨龟鹿二仙胶含药血清对大鼠膝关节软骨细胞肥大分化的调控机制。方法取4周龄SPF级SD大鼠膝关节软骨细胞进行传代培养,将第三代生长良好的软骨细胞,设为空白组;采用含10 ng/mL IL-1β的10%FBS/DMEM培养上述第... 目的基于IHH-Gli信号通路探讨龟鹿二仙胶含药血清对大鼠膝关节软骨细胞肥大分化的调控机制。方法取4周龄SPF级SD大鼠膝关节软骨细胞进行传代培养,将第三代生长良好的软骨细胞,设为空白组;采用含10 ng/mL IL-1β的10%FBS/DMEM培养上述第三代软骨细胞12 h后制备大鼠膝关节肥大软骨细胞模型,模型鉴定成功后随机分为模型组、龟鹿组、抑制剂组。空白组、模型组均采用含10%大鼠空白血清DMEM进行培养,龟鹿组和抑制剂组分别采用含10%龟鹿二仙胶大鼠含药血清和含10 nmol/L环巴胺的10%大鼠空白血清DMEM进行培养。培养72 h后在光镜下观察4组软骨细胞形态变化,qRT-PCR法检测4组印度刺猬(IHH)、膜受体Smoothened(Smo)、胶质细胞瘤转录因子(Gli)、Runt相关转录因子2(Runx2)、X型胶原α1(Col10A1)、基质金属蛋白酶13(MMP-13)mRNA相对表达水平,采用Western blot检测等4组IHH、Smo、Gli蛋白表达量。结果与空白组比较,模型组细胞不规则形态增加,细胞间隙变大、数量变少,细胞碎片、悬浮死细胞增多;与模型组比较,龟鹿组、抑制剂组细胞不规则形态减少,细胞间隙变小、数量增多,细胞碎片、悬浮死细胞减少。与空白组比较,模型组、龟鹿组和抑制剂组IHH、Smo、Gli、Runx2、Col10A1、MMP-13 mRNA相对表达水平及IHH、Smo、Gli蛋白表达量均有一定程度提高(P均<0.05);与模型组比较,龟鹿组和抑制剂组IHH、Smo、Gli、Runx2、Col10A1、MMP-13 mRNA相对表达水平及IHH、Smo、Gli蛋白表达量均显著下调(P均<0.05)。结论龟鹿二仙胶含药血清可调节大鼠膝关节肥大软骨细胞的分解合成平衡,改善肥大软骨细胞的失稳态环境,其调控机制可能与IHH-Gli信号通路相关。 展开更多
关键词 膝关节软骨细胞 肥大细胞分化 凋亡 骨关节炎 龟鹿二仙胶
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Altered Gene Expression in Articular Chondrocytes of Smad3^(ex8/ex8) Mice, Revealed by Gene Profiling Using Microarrays 被引量:2
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作者 王浩 张继帅 +1 位作者 孙强 杨晓 《Journal of Genetics and Genomics》 SCIE CAS CSCD 北大核心 2007年第8期698-708,共11页
It has been previously reported that small mother against decapentaplegic 3 (Smad3) gene knockout (Smad3^ex8/ex8) mice displays phenotypes similar to human osteoarthritis, as characterized by abnormal hypertrophic... It has been previously reported that small mother against decapentaplegic 3 (Smad3) gene knockout (Smad3^ex8/ex8) mice displays phenotypes similar to human osteoarthritis, as characterized by abnormal hypertrophic differentiation of articular chondrocytes. To further clarify the crucial target genes that mediate transformation growth factor-β (TGF-β)/Smad3 signals on articular chondrocytes differentiation and investigate the underlying molecular mechanism of osteoarthritis, microarrays were used to perform comparative transcriptional profiling in the articular cartilage between Smad3^ex8/ex8and wild-type mice on day five after birth. The gene profding results showed that the activity of bone morphogenetic protein (BMP) and TGF-β/cell division cycle 42 (Cdc42) signaling pathways were enhanced in Smad3^ex8/ex8 chondrocytes. Moreover, there was altered gene expression in growth hormone/insulin-like growth factor 1 (Igfl) axis and fibroblast growth factor (Fgf) signaling pathway. Notably, protein synthesis related genes and electron transport chain related genes were upregulated in Smad3^ex8/ex8 chondrocytes, implying that accelerated protein synthesis and enhanced cellular respiration might contribute to hypertrophic differentiation of articular chondrocytes and the pathogenesis of osteoarthritis. 展开更多
关键词 TGF-β SMAD3 articular chondrocytes hypertrophic differentiation OSTEOARTHRITIS MICROARRAY
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