Adiponectin Attenuates Lipopolysaccharide-induced Apoptosis by Regulating the Cx43/PI3K/AKT Pathway.
Adiponectin Attenuates Lipopolysaccharide-induced Apoptosis by Regulating the Cx43/PI3K/AKT Pathway.
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脂联素通过调节 Cx43/PI3K/AKT 途径减弱脂多糖诱导的细胞凋亡
DOI：
10.3389/fphar.2021.644225
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发表时间：
2021
影响因子：
5.6
通讯作者：
Ma K
中科院分区：
文献类型：
--
作者：
Liu L;Yan M;Yang R;Qin X;Chen L;Li L;Si J;Li X;Ma K
心肌细胞凋亡是导致心肌功能障碍的重要因素。脂联素（APN）具有心肌保护作用。研究表明，连接蛋白43（Cx43）和磷脂酰肌醇-3-激酶（PI 3 K）/蛋白激酶B（AKT）信号通路在心脏中发挥重要作用，但APN是否通过调节这些通路发挥保护作用尚不清楚。本研究旨在探讨APN是否通过调节Cx43和PI 3 K/AKT信号通路发挥抗脂多糖（LPS）诱导的心肌细胞凋亡作用。此外，本研究还探讨了Cx43与PI 3 K/AKT信号通路的关系。体外实验：LPS处理H9 c2细胞24 h前，先用APN预处理2 h。CCK-8法检测APN对H9 c2细胞的杀伤作用。Western blot检测Bax、Bcl 2、cleaved caspase-3、cleaved caspase-9、Cx43、PI 3 K、p-PI 3 K、AKT和p-AKT蛋白表达水平，流式细胞术检测细胞凋亡率。APN可减弱LPS诱导的细胞毒性。LPS上调Bax，切割caspase-3和切割caspase-9和下调Bcl-2在H9 c2细胞;然而，这些影响被APN减弱。此外，LPS上调Cx43表达，APN下调Cx43表达并激活PI 3 K/AKT信号通路。LPS可诱导H9 c2细胞凋亡，并抑制PI 3 K/AKT信号通路，Gap 26（Cx43抑制剂）可减弱这种作用。此外，APN表达的保存被LY 294002（PI 3 K/AKT信号通路抑制剂）逆转。体内实验：C57 BL/6 J小鼠腹腔注射LPS建立脓毒症模型，腹腔注射APN。Western blot法检测心肌组织中Bax、Bcl 2、Caspase-3和Cx43蛋白的表达，免疫组化法检测Cx43在心肌组织中的表达和定位。LPS上调Bax和切割caspase-3和下调Bcl 2在脓毒症，然而，这些影响被APN减弱。此外，在脓毒症心肌组织中，Cx43的表达上调，APN下调Cx43的表达。总之，无论是在体外还是在体内，数据表明APN可以通过修饰Cx43和PI 3 K/AKT信号通路来保护脓毒症期间LPS诱导的细胞凋亡。
Cardiomyocyte apoptosis is a crucial factor leading to myocardial dysfunction. Adiponectin (APN) has a cardiomyocyte-protective impact. Studies have shown that the connexin43 (Cx43) and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathways play an important role in the heart, but whether APN plays a protective role by regulating these pathways is unclear. Our study aimed to confirm whether APN protects against lipopolysaccharide (LPS)-induced cardiomyocyte apoptosis and to explore whether it plays an important role through regulating the Cx43 and PI3K/AKT signaling pathways. In addition, our research aimed to explore the relationship between the Cx43 and PI3K/AKT signaling pathways. In vitro experiments: Before H9c2 cells were treated with LPS for 24 h, they were pre-treated with APN for 2 h. The cytotoxic effect of APN on H9c2 cells was evaluated by a CCK-8 assay. The protein levels of Bax, Bcl2, cleaved caspase-3, cleaved caspase-9, Cx43, PI3K, p-PI3K, AKT and p-AKT were evaluated by Western blot analysis, and the apoptosis rate was evaluated by flow cytometry. APN attenuated the cytotoxicity induced by LPS. LPS upregulated Bax, cleaved caspase-3 and cleaved caspase-9 and downregulated Bcl2 in H9c2 cells; however, these effects were attenuated by APN. In addition, LPS upregulated Cx43 expression, and APN downregulated Cx43 expression and activated the PI3K/AKT signaling pathway. LPS induced apoptosis and inhibited PI3K/AKT signaling pathway in H9c2 cells, and these effects were attenuated by Gap26 (a Cx43 inhibitor). Moreover, the preservation of APN expression was reversed by LY294002 (a PI3K/AKT signaling pathway inhibitor). In vivo experiments: In C57BL/6J mice, a sepsis model was established by intraperitoneal injection of LPS, and APN was injected into enterocoelia. The protein levels of Bax, Bcl2, cleaved caspase-3, and Cx43 were evaluated by Western blot analysis, and immunohistochemistry was used to detect Cx43 expression and localization in myocardial tissue. LPS upregulated Bax and cleaved caspase-3 and downregulated Bcl2 in sepsis; however, these effects were attenuated by APN. In addition, the expression of Cx43 was upregulated in septic myocardial tissue, and APN downregulated Cx43 expression in septic myocardial tissue. In conclusion, both in vitro and in vivo, the data demonstrated that APN can protect against LPS-induced apoptosis during sepsis by modifying the Cx43 and PI3K/AKT signaling pathways.
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影响因子：
3.7
作者：
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