小檗碱抑制钙盐诱导主动脉瓣膜间质细胞成骨分化  

Berberine inhibits osteogenic differentiation of aortic valve interstitial cells induced by osteogenic induction medium

作　　者：黄琴 王玥 李晓蓉 向熠 施琼[1] HUANG Qin;WANG Yue;LI Xiao-rong;XIANG Yi;SHI Qiong(Key Laboratory of Clinical Laboratory Diagnostics,Ministry of Education,College of Laboratory Medicine,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国中药杂志》2022年第11期3059-3065,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81672103)。

摘　　要：该研究主要探讨了小檗碱对钙盐诱导的瓣膜间质细胞成骨分化的作用及其机制,为钙化性主动脉瓣膜疾病的临床治疗提供新的理论思路。使用Western blot检测3例钙化瓣膜组织和1例正常瓣膜组织相关成骨和纤维化指标的表达情况;分离猪瓣膜间质细胞后,MTT实验检测不同浓度小檗碱对瓣膜间质细胞活力的影响并确定小檗碱的最佳浓度范围;使用钙盐培养基培养瓣膜间质细胞,并加入不同浓度的小檗碱处理,Western blot和q-PCR检测不同浓度小檗碱对瓣膜间质细胞纤维化和成骨相关指标表达情况的影响;ALP染色和茜素红S染色分别检测小檗碱对钙盐诱导的瓣膜间质细胞早期及晚期成骨分化能力的影响;Western blot检测小檗碱对不同时间段ERK1/2磷酸化水平的影响,并加入ERK1/2磷酸化抑制剂PD98059进行验证。结果显示相关成骨指标和纤维化指标在钙化主动脉瓣膜组织相较于正常组织明显上调;小檗碱可以明显降低钙盐诱导的瓣膜间质细胞相关纤维化和成骨指标的上调,降低ALP活性和钙盐沉积,降低ERK1/2磷酸化水平;并且PD98059处理同样降低了钙盐诱导的瓣膜间质细胞相关纤维化和成骨指标的上调。该研究表明小檗碱可以抑制瓣膜间质细胞向肌成纤维细胞或成骨细胞样细胞分化,其机制可能与抑制ERK1/2信号通路有关。This research focused on the effect and mechanism of berberine on osteogenic differentiation of valve interstitial cells(VICs) induced by osteogenic induction medium, in order to provide new insights into the clinical treatment of calcified aortic valve disease. The expression of osteogenic and fibrotic makers in three cases of calcified valve tissues and one case of normal control was assayed by Western blot. After the porcine aortic VICs were isolated, the effects of different concentrations of berberine on their viability were examined by MTT assay for determining the optimal concentration range. VICs were cultured in osteogenic induction medium and treated with different concentrations of berberine. Western blot and q-PCR were conducted to detect the effects of berberine on the expression of osteogenic and fibrotic makers in VICs. The effects of berberine on osteogenic differentiation of VICs in the early and late stages were separately measured by ALP staining and alizarin red S staining. The effects of berberine on the phosphorylation of ERK1/2 at different time points were assayed by Western blot. And PD98059, an inhibitor of ERK1/2, was added for verification. The results suggested that related osteogenic and fibrotic makers were significantly up-regulated in calcified valve tissues as compared with those in the normal control. The up-regulated fibrosis and osteogenic makers of VICs under osteogenic conditions were reversed by berberine and the ALP activity and calcium deposition in VICs were also reduced obviously. The level of ERK1/2 phosphorylation was decreased. Similarly, the osteogenic and fibrotic makers of VICs induced by osteogenic induction medium were lowered by PD98059. This study has confirmed that berberine is able to inhibit the differentiation of VICs into myofibroblasts or osteoblast-like cells, which may be associated with the inhibition of ERK1/2 signaling pathway.

关 键 词：小檗碱 钙化性主动脉瓣膜病 瓣膜间质细胞 成骨分化 

分 类 号：R285[医药卫生—中药学]