迷迭香酸通过Vimentin/NLRP3影响人肝癌HepG2细胞的增殖及凋亡
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摘要
目的观察迷迭香酸对人肝癌HepG2细胞增殖及凋亡的影响,并探讨Vimentin/NLRP3在此过程中的作用。方法不同浓度迷迭香酸(25、50、100μmol·L~(-1))作用于HepG2细胞, MTT法检测细胞增殖;TUNEL法检测细胞凋亡;Western blot检测相关蛋白的表达水平。结果迷迭香酸能抑制HepG2细胞的增殖,促进其凋亡,且呈浓度依赖性。与对照组相比,迷迭香酸组能明显上调caspase-3的表达,降低Bcl-2的表达(P<0.05),其作用机制与下调Vimentin/NLRP3信号通路有关。结论迷迭香酸能抑制HepG2细胞增殖并诱导细胞凋亡,其作用主要是通过下调Vimentin/NLRP3蛋白的表达来实现的。
Aim To investigate the effect of rosmarinic acid on the proliferation and apoptosis of HepG2 cells and the role of Vimentin/NLRP3 in this process. Methods HepG2 cells were treated by different concentrations of rosmarinic acid(25, 50, 100 μmol·L~(-1)). Cell proliferation was assayed by MTT method. Apoptosis was detected by TUNEL, and the expressions of NLRP3 protein and Vimentin were measured by Western blot. The expression of TNF-α and IL-β were investigated by ELISA. Results Compared with control group, rosmarinic acid at 25, 50, 100 μmol·L~(-1) could inhibit the proliferation and induced apoptosis of HepG2 cells in a dose-depended manner(P<0.05). Rosmarinic acid could increase the the expression levels of caspase-3, but significantly suppressed the of Bcl-2 expression levels(P<0.01). The mechanism might be related to down-regulating the Vimentin/NLRP3 signaling pathway. Conclusions Rosmarinic acid can synergistically inhibit the proliferation of HepG2 cells and induce its apoptosis in a dose-dependent manner by down-regulating the protein level of NLRP3 and Vimentin.
Aim To investigate the effect of rosmarinic acid on the proliferation and apoptosis of HepG2 cells and the role of Vimentin/NLRP3 in this process. Methods HepG2 cells were treated by different concentrations of rosmarinic acid(25, 50, 100 μmol·L~(-1)). Cell proliferation was assayed by MTT method. Apoptosis was detected by TUNEL, and the expressions of NLRP3 protein and Vimentin were measured by Western blot. The expression of TNF-α and IL-β were investigated by ELISA. Results Compared with control group, rosmarinic acid at 25, 50, 100 μmol·L~(-1) could inhibit the proliferation and induced apoptosis of HepG2 cells in a dose-depended manner(P<0.05). Rosmarinic acid could increase the the expression levels of caspase-3, but significantly suppressed the of Bcl-2 expression levels(P<0.01). The mechanism might be related to down-regulating the Vimentin/NLRP3 signaling pathway. Conclusions Rosmarinic acid can synergistically inhibit the proliferation of HepG2 cells and induce its apoptosis in a dose-dependent manner by down-regulating the protein level of NLRP3 and Vimentin.
引文
[1] 陈莉莉,杨雪梅,谌贝贝,等.肝X受体激活对全脑缺血/再灌注小鼠海马神经干细胞增殖及认知功能的影响[J].中国药理学通报,2018,34(2):185-90.
[1] Chen L L,Yang X M, Chen B B, et al. Effect of liver X receptor activation on hippocampal neural stem cell proleferation and cognitive function in global cerebral ischemia/reperfusion mice[J].Chin Pharmacol Bull, 2018,34(2):185-90.
[2] Cheng J, Zhong Y, Chen S, et al. Gab2 mediates hepatocellular carcinogenesis by integrating multiple signaling pathways[J].FASEB,2017,31(12):5530-42.
[3] 郭 峰,朱炳阳,迟秀玲,等.迷迭香酸抗氧化氢诱导血管平滑肌细胞凋亡作用的研究[J]. 中国药理学通报,2007,23(3):365-7.
[3] Guo F,Zhu B Y,Chi X L,et al. Inhibition of rosmarinic acid on the apoptosis of vascular smooth muscle cells induced by hydrogen peroxide[J].Chin Pharmacol Bull, 2007,23(3):365-7.
[4] Hahn H J,Kim K B,An I S,et al. Protective effects of rosmarinic acid against hydrogen peroxide-induced cellular senescence and inflammatory response in normal human dermal fibroblasts[J]. Mol Med Rep, 2017,16(6): 9763-9.
[5] Hasanein P, Sharifi M. Effects of rosmarinic acid on actetaminophen-induced hepatotoxicity in male Wistar rats[J]. Pharm Biol, 2017,55(1): 1809-16.
[6] Sesarman A,Tefas L,Sylvester B,et al.Anti-angiogenic and anti-inflammatory effects of long-circulating liposomes co-encapsulating curcumin and doxorubicin on C26 murine colon cancer cells[J]. Pharmacol Rep, 2017, 70(2):331-9.
[7] Satelli A,Li S. Vimentin in cancer and its potential as a molecular target for cancer therapy[J]. Cell Mol Life Sci, 2011,68(18): 3033.
[8] 杨艺萱,郭文静,马承旭,等.血根碱通过诱导活性氧促进HepG2细胞凋亡的机制研究[J]. 中国药理学通报, 2018,34(3):370-5.
[8] Yang Y X,Guo W J,Ma C X,et al.Sanguinarune promotes apoptosis of HepG2 cells by inducing ROS[J]. Chin Pharmacol Bull, 2018,34(3):370-5.
[9] Coelho V R,Viau C M,Staub R B,et al. Rosmarinic acid attenuates the activation of murine microglial N9 cells through the downregulation of inflammatory cytokines and cleaved caspase-3[J]. Neuroimmunomodulation, 2017,24(3):171-81.
[10] 杨沛霖,高 玲,林彩霞,等.迷迭香酸对鼻咽癌细胞增殖、凋亡的影响及其作用机制[J]. 山东医药, 57(36):1-5.
[10] Yang P L,Gao L,Lin C X,et al. Rosmarinic acid suppresses cell proliferation and induced apoptosis of human nasopharyngeal carcinoma cell line CNE-1[J]. Shandong Med J, 57(36):1-5.
[11] 任 为,牟宜双,可 熊. 迷迭香酸通过诱导凋亡及抑制NF-κB信号通路发挥抗肝癌作用的机制研究[J]. 中药药理与临床, 2016, 32(2):31-4.
[11] Ren W,Mou Y S, Ke X. Sodium rosmarinate can inhibit proliferation of hepatocellular carcinoma in mice via inducing apoptosis and downregulation of NF-κB pathway[J]. Pharmacol Clin Chin Mater Med, 2016,32(2):31-4.
[12] 苏 杰, 姚 杨, 朱星枚,成碧萍. 迷迭香酸干预下HBX基因对人肝癌细胞凋亡的影响[J]. 西部医学, 2015,27(2):178-81.
[12] Su J, Yao Y, Zhu X M, et al. Effect of HBx gene on human HepG2 cell after rosmarinic acid intervention[J]. Med J West China, 2015,27(2):178-81.
[13] 卓少元,谢金玲.健脾益气方含药血清下调Vimentin 蛋白水平对人肝癌细胞SMMC-7721的影响[J]. 时珍国医国药, 2017,28(1):55-9.
[13] Zhuo S Y, Xie J L. Effects on SMMC-7721 cell behavior due to down-regulation of Vimentin expression by serum containing Jianpi Yiqi decpction[J]. Lishizhen Med Mater Med Res, 2017,28(1):55-9.
[14] 王红光,贾 强,张述升,等.波形蛋白、NF-κB促进胶质瘤侵袭性的表达相关性及意义[J]. 中国微侵袭神经外科杂志, 2015,20(6):276-8.
[14] Wang H G,Jia Q,Zhang S S,et al.Expression and significance of vimentin and NF-κB in invasion glioma[J]. Chin J Minim Invasion Neurosurg, 2015,20(6):276-8.
[1] Chen L L,Yang X M, Chen B B, et al. Effect of liver X receptor activation on hippocampal neural stem cell proleferation and cognitive function in global cerebral ischemia/reperfusion mice[J].Chin Pharmacol Bull, 2018,34(2):185-90.
[2] Cheng J, Zhong Y, Chen S, et al. Gab2 mediates hepatocellular carcinogenesis by integrating multiple signaling pathways[J].FASEB,2017,31(12):5530-42.
[3] 郭 峰,朱炳阳,迟秀玲,等.迷迭香酸抗氧化氢诱导血管平滑肌细胞凋亡作用的研究[J]. 中国药理学通报,2007,23(3):365-7.
[3] Guo F,Zhu B Y,Chi X L,et al. Inhibition of rosmarinic acid on the apoptosis of vascular smooth muscle cells induced by hydrogen peroxide[J].Chin Pharmacol Bull, 2007,23(3):365-7.
[4] Hahn H J,Kim K B,An I S,et al. Protective effects of rosmarinic acid against hydrogen peroxide-induced cellular senescence and inflammatory response in normal human dermal fibroblasts[J]. Mol Med Rep, 2017,16(6): 9763-9.
[5] Hasanein P, Sharifi M. Effects of rosmarinic acid on actetaminophen-induced hepatotoxicity in male Wistar rats[J]. Pharm Biol, 2017,55(1): 1809-16.
[6] Sesarman A,Tefas L,Sylvester B,et al.Anti-angiogenic and anti-inflammatory effects of long-circulating liposomes co-encapsulating curcumin and doxorubicin on C26 murine colon cancer cells[J]. Pharmacol Rep, 2017, 70(2):331-9.
[7] Satelli A,Li S. Vimentin in cancer and its potential as a molecular target for cancer therapy[J]. Cell Mol Life Sci, 2011,68(18): 3033.
[8] 杨艺萱,郭文静,马承旭,等.血根碱通过诱导活性氧促进HepG2细胞凋亡的机制研究[J]. 中国药理学通报, 2018,34(3):370-5.
[8] Yang Y X,Guo W J,Ma C X,et al.Sanguinarune promotes apoptosis of HepG2 cells by inducing ROS[J]. Chin Pharmacol Bull, 2018,34(3):370-5.
[9] Coelho V R,Viau C M,Staub R B,et al. Rosmarinic acid attenuates the activation of murine microglial N9 cells through the downregulation of inflammatory cytokines and cleaved caspase-3[J]. Neuroimmunomodulation, 2017,24(3):171-81.
[10] 杨沛霖,高 玲,林彩霞,等.迷迭香酸对鼻咽癌细胞增殖、凋亡的影响及其作用机制[J]. 山东医药, 57(36):1-5.
[10] Yang P L,Gao L,Lin C X,et al. Rosmarinic acid suppresses cell proliferation and induced apoptosis of human nasopharyngeal carcinoma cell line CNE-1[J]. Shandong Med J, 57(36):1-5.
[11] 任 为,牟宜双,可 熊. 迷迭香酸通过诱导凋亡及抑制NF-κB信号通路发挥抗肝癌作用的机制研究[J]. 中药药理与临床, 2016, 32(2):31-4.
[11] Ren W,Mou Y S, Ke X. Sodium rosmarinate can inhibit proliferation of hepatocellular carcinoma in mice via inducing apoptosis and downregulation of NF-κB pathway[J]. Pharmacol Clin Chin Mater Med, 2016,32(2):31-4.
[12] 苏 杰, 姚 杨, 朱星枚,成碧萍. 迷迭香酸干预下HBX基因对人肝癌细胞凋亡的影响[J]. 西部医学, 2015,27(2):178-81.
[12] Su J, Yao Y, Zhu X M, et al. Effect of HBx gene on human HepG2 cell after rosmarinic acid intervention[J]. Med J West China, 2015,27(2):178-81.
[13] 卓少元,谢金玲.健脾益气方含药血清下调Vimentin 蛋白水平对人肝癌细胞SMMC-7721的影响[J]. 时珍国医国药, 2017,28(1):55-9.
[13] Zhuo S Y, Xie J L. Effects on SMMC-7721 cell behavior due to down-regulation of Vimentin expression by serum containing Jianpi Yiqi decpction[J]. Lishizhen Med Mater Med Res, 2017,28(1):55-9.
[14] 王红光,贾 强,张述升,等.波形蛋白、NF-κB促进胶质瘤侵袭性的表达相关性及意义[J]. 中国微侵袭神经外科杂志, 2015,20(6):276-8.
[14] Wang H G,Jia Q,Zhang S S,et al.Expression and significance of vimentin and NF-κB in invasion glioma[J]. Chin J Minim Invasion Neurosurg, 2015,20(6):276-8.