hPMSCs protects against D-galactose-induced oxidative damage of CD4(+) T cells through activating Akt-mediated Nrf2 antioxidant signaling.
hPMSCs protects against D-galactose-induced oxidative damage of CD4(+) T cells through activating Akt-mediated Nrf2 antioxidant signaling.
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HPMSC通过激活Akt介导的NRF2抗氧化剂信号传导来预防D-半乳糖诱导的CD4（+）T细胞的氧化损伤。
DOI：
10.1186/s13287-020-01993-0
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发表时间：
2020-11-04
影响因子：
7.5
通讯作者：
Luan X
中科院分区：
文献类型：
--
作者：
Xiong Y;Wang Y;Zhang J;Zhao N;Zhang H;Zhang A;Zhao D;Yu Z;Yin Y;Song L;Xiong Y;Luan X
骨髓间充质干细胞（MSC）被认为是一种再生治疗急性和慢性疾病的方法。然而，MSC是否调节CD 4 + T细胞的抗氧化代谢并减弱免疫衰老仍不清楚。在这里，我们报道了hPMSCs在衰老相关的CD 4 + T细胞衰老中的保护作用，并使用d-gal诱导的小鼠衰老模型确定了潜在的机制。在体内实验中，将40只8周龄雄性C57 BL/6小鼠随机分为4组：对照组、d-gal组、hPMSC组和PBS组。在体外实验中，使用初始CD 4 + T细胞分离试剂盒II制备人初始CD 4 + T（CD 4CD 45 RA）细胞，并用Akt抑制剂LY 294002和Nrf 2抑制剂ML 385预处理。然后，将分离的幼稚CD 4 + T细胞与hPMSC在不存在或存在抗CD 3/CD 28 Dynabeads和IL-2作为促有丝分裂刺激物的情况下共培养72小时。流式细胞仪检测细胞内ROS的变化。抗氧化酶超氧化物歧化酶，谷胱甘肽过氧化物酶和过氧化氢酶的活性测定比色分析。SA-β-gal染色检测衰老T细胞。采用Western blotting、RT-PCR和共聚焦显微镜检测衰老相关蛋白的表达。我们发现hPMSC处理显著降低了衰老CD 4 + T细胞中的ROS水平、SA-β-gal阳性细胞数量、衰老相关分泌表型（IL-6和OPN）表达以及衰老相关蛋白（P16和P21）表达。此外，hPMSC处理有效地上调Nrf 2核转位和衰老CD 4 + T细胞中下游靶基因（HO-1、CAT、GCLC和NQO 1）的表达。此外，体外研究显示，hPMSC通过上调Akt/GSK-3β/Fyn通路以激活Nrf 2功能来减弱CD 4 + T细胞衰老。相反，hPMSC的抗氧化作用在衰老的CD 4 + T细胞中被Akt抑制剂LY 294002和Nrf 2抑制剂ML 385阻断。我们的研究结果表明，hPMSC通过激活Nrf 2介导的抗氧化防御来减弱d-gal诱导的CD 4 + T细胞衰老，并且hPMSC上调Nrf 2是通过Akt/GSK-3β/Fyn途径调节的。
Mesenchymal stem cells (MSCs) were considered a regenerative therapeutic approach in both acute and chronic diseases. However, whether MSCs regulate the antioxidant metabolism of CD4+ T cells and weaken immunosenescence remains unclear. Here, we reported the protective effects of hPMSCs in aging-related CD4+ T cell senescence and identified the underlying mechanisms using a d-gal-induced mouse aging model. In vivo study, 40 male C57BL/6 mice (8 weeks) were randomly divided into four groups: control group, d-gal group, hPMSC group, and PBS group. In in vitro experiment, human naive CD4+ T (CD4CD45RA) cells were prepared using a naive CD4+ T cell isolation kit II and pretreated with the Akt inhibitor LY294002 and Nrf2 inhibitor ML385. Then, isolated naive CD4+ T cell were co-cultured with hPMSCs for 72 h in the absence or presence of anti-CD3/CD28 Dynabeads and IL-2 as a mitogenic stimulus. Intracellular ROS changes were detected by flow cytometry. The activities of the antioxidant enzymes superoxide dismutase, glutathione peroxidase, and catalase were measured by colorimetric analysis. The senescent T cells were detected SA-β-gal stain. The expression of aging-related proteins was detected by Western blotting, RT-PCR, and confocal microscopy. We found that hPMSC treatment markedly decreased the ROS level, SA-β-gal-positive cells number, senescence-associated secretory phenotype (IL-6 and OPN) expression, and aging-related protein (P16 and P21) expression in senescent CD4+ T cells. Furthermore, hPMSC treatment effectively upregulated Nrf2 nuclear translocation and the expression of downstream target genes (HO-1, CAT, GCLC, and NQO1) in senescent CD4+ T cells. Moreover, in vitro studies revealed that hPMSCs attenuated CD4+ T cell senescence by upregulating the Akt/GSK-3β/Fyn pathway to activate Nrf2 functions. Conversely, the antioxidant effects of hPMSCs were blocked by the Akt inhibitor LY294002 and Nrf2 inhibitor ML385 in senescent CD4+ T cells. Our results indicate that hPMSCs attenuate d-gal-induced CD4+ T cell senescence by activating Nrf2-mediated antioxidant defenses and that upregulation of Nrf2 by hPMSCs is regulated via the Akt/GSK-3β/Fyn pathway.
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影响因子：
16.6
作者：
Kuwahara, Makoto;Suzuki, Junpei;Tofukuji, Soichi;Yamada, Takeshi;Kanoh, Makoto;Matsumoto, Akira;Maruyama, Saho;Kometani, Kohei;Kurosaki, Tomohiro;Ohara, Osamu;Nakayama, Toshinori;Yamashita, Masakatsu
通讯作者：
Yamashita, Masakatsu
Nrf2 的上调参与 FGF21 介导的非诺贝特对 1 型糖尿病肾病的保护作用。
影响因子：
7.4
作者：
Cheng Y;Zhang J;Guo W;Li F;Sun W;Chen J;Zhang C;Lu X;Tan Y;Feng W;Fu Y;Liu GC;Xu Z;Cai L
通讯作者：
Cai L
影响因子：
7.7
作者：
Lian G;Gnanaprakasam JR;Wang T;Wu R;Chen X;Liu L;Shen Y;Yang M;Yang J;Chen Y;Vasiliou V;Cassel TA;Green DR;Liu Y;Fan TW;Wang R
通讯作者：
Wang R
影响因子：
7.3
作者：
Rashida Gnanaprakasam JN;Wu R;Wang R
通讯作者：
Wang R
影响因子：
5.8
作者：
Bryan, Holly K.;Olayanju, Adedamola;Park, B. Kevin
通讯作者：
Park, B. Kevin