Li Xiaoou, Pan Feng, He Bing, Fang Chengzhi
Department of Neonatology, Renmin Hospital, Wuhan University, Wuhan, Hubei 430060, People's Republic of China.
Department of Orthopedics, Renmin Hospital, Wuhan University, Wuhan, Hubei 430060, People's Republic of China.
Open Life Sci. 2021 Aug 27;16(1):856-866. doi: 10.1515/biol-2021-0081. eCollection 2021.
The present research was designed to examine the effects of disintegrin metalloproteinases 10 (ADAM10) on the doxorubicin (DOX)-induced dilated cardiomyopathy (DCM) and the mechanisms involved, with a focus on ADAM10-dependent cleavage of N-cadherin. The present study constructed recombinant lentiviral vectors expressing short hairpin RNA (shRNA) targeting the ADAM10 gene. H9C2 cells were treated with the recombinant lentivirus or GI254023 (an ADAM10 inhibitor). The expression level of N-cadherin and its C-terminal fragment1 (CTF1) was tested by western blotting and flow cytometry. The adhesion ability was analyzed using a plate adhesion model. Cardiac function and morphology were assessed in control and lentivirus-transfected rats with or without DOX treatment. The inhibition of ADAM10 activity significantly increased the expression of full-length N-cadherin on the cellular surface and reduced CTF1 generation and . The adhesion ability was also increased in ADAM10-knockdown H9C2 cells. Furthermore, DOX-induced myocardial dysfunction was ameliorated in rats transfected with ADAM10-shRNA lentivirus. These findings demonstrated that ADAM10 specifically cleaves N-cadherin in cardiomyocytes. ADAM10-induced N-cadherin cleavage results in changes in the adhesive behavior of cells. Therefore, ADAM10 may serve as a therapeutic target to reverse cardiac remodeling in DCM.
本研究旨在探讨解整合素金属蛋白酶10（ADAM10）对阿霉素（DOX）诱导的扩张型心肌病（DCM）的影响及其相关机制，重点关注ADAM10依赖性N-钙黏蛋白的裂解。本研究构建了表达靶向ADAM10基因的短发夹RNA（shRNA）的重组慢病毒载体。用重组慢病毒或GI254023（一种ADAM10抑制剂）处理H9C2细胞。通过蛋白质印迹法和流式细胞术检测N-钙黏蛋白及其C末端片段1（CTF1）的表达水平。使用平板黏附模型分析黏附能力。在对照和经慢病毒转染的大鼠中评估心脏功能和形态，这些大鼠接受或未接受DOX处理。抑制ADAM10活性显著增加了细胞表面全长N-钙黏蛋白的表达，并减少了CTF1的产生。在ADAM10基因敲低的H9C2细胞中，黏附能力也有所增加。此外，在转染了ADAM10-shRNA慢病毒的大鼠中，DOX诱导的心肌功能障碍得到改善。这些发现表明，ADAM10在心肌细胞中特异性裂解N-钙黏蛋白。ADAM10诱导的N-钙黏蛋白裂解导致细胞黏附行为的改变。因此，ADAM10可能作为逆转DCM心脏重塑的治疗靶点。
