黄芪总黄酮通过调控miR-378对高糖诱导的HK-2细胞炎症因子水平的影响
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摘要
目的研究黄芪总黄酮对高糖诱导的HK-2细胞炎症水平的影响,并探讨其与miR-378之间的关系。方法运用MTT法检测黄芪总黄酮(0、50、100、150 mmol/L)与HG(25 mmol/L)共处理的HK-2细胞的增殖;将miR-NC+HG组(转染miRNC)、miR-378+HG组(转染miR-378 mimics)均以脂质体法转染至25 mmol/L HG处理的HK-2细胞;用ELISA法检测各组细胞中IL-6、TNF-α、IgA、IgM、IgG含量;用qRT-PCR检测各组细胞中miR-378表达。结果与Control组相比,各个浓度的TFA处理的HK-2细胞的增殖率均显著升高,IL-6、TNF-α、IgA、IgM含量均明显降低,IgG、miR-378明显升高(P<0.05);过表达miR-378可使HK-2细胞的增殖率均显著升高,IL-6、TNF-α、IgA、IgM含量显著降低,IgG含量显著升高。结论黄芪总黄酮可降低高糖诱导的HK-2细胞炎症水平,其机制与直接上调miR-378有关,将可为黄芪总黄酮治疗糖尿病肾病提供理论依据。
This study was performed to study the effect of Astragalus total flavonoids(TFA) on high glucose(HG)-induced inflammation level of HK-2 cells and explore its relationship with miR-378. MTT assay was used to detect the proliferation of HK-2 cells co-treated with total flavonoids(0, 50, 100, 150 mmol/L) and HG(25 mmol/L);miR-NC + HG group(transfected miR-NC) and miR-378 + HG group(transfected miR-378 mimics) were all transfected into 25 mmol/L HG-treated HK-2 cells by liposome method. The levels of IL-6, TNF-α, IgA, IgM, and IgG were detected by ELISA; q RT-PCR was used to detect miR-378 expression in each group of cells. Compared with the control group, the proliferation rate of HK-2 cells treated with TFA in each concentration was significantly increased, the levels of IL-6, TNF-α, IgA, and IgM were significantly decreased, while the levels of IgG and miR-378 were significantly increased(P<0.05). Overexpression of miR-378 significantly increased the proliferation rate of HK-2 cells, significantly decreased the levels of IL-6, TNF-α, IgA, and IgM, while increased the level of IgG significantly. Taken together, Astragalus total flavonoids can reduce the inflammation level of HK-2 cells induced by high glucose, and the mechanism may be related to the direct up-regulation of miR-378, which will provide a theoretical basis for the treatment of diabetic nephropathy with Astragalus total flavonoids.
This study was performed to study the effect of Astragalus total flavonoids(TFA) on high glucose(HG)-induced inflammation level of HK-2 cells and explore its relationship with miR-378. MTT assay was used to detect the proliferation of HK-2 cells co-treated with total flavonoids(0, 50, 100, 150 mmol/L) and HG(25 mmol/L);miR-NC + HG group(transfected miR-NC) and miR-378 + HG group(transfected miR-378 mimics) were all transfected into 25 mmol/L HG-treated HK-2 cells by liposome method. The levels of IL-6, TNF-α, IgA, IgM, and IgG were detected by ELISA; q RT-PCR was used to detect miR-378 expression in each group of cells. Compared with the control group, the proliferation rate of HK-2 cells treated with TFA in each concentration was significantly increased, the levels of IL-6, TNF-α, IgA, and IgM were significantly decreased, while the levels of IgG and miR-378 were significantly increased(P<0.05). Overexpression of miR-378 significantly increased the proliferation rate of HK-2 cells, significantly decreased the levels of IL-6, TNF-α, IgA, and IgM, while increased the level of IgG significantly. Taken together, Astragalus total flavonoids can reduce the inflammation level of HK-2 cells induced by high glucose, and the mechanism may be related to the direct up-regulation of miR-378, which will provide a theoretical basis for the treatment of diabetic nephropathy with Astragalus total flavonoids.
引文
[1]Xu Y,Wang L,He J,et al.Prevalence and control of diabetes in Chinese adults[J].JAMA,2013,310:948-959.
[2]Yu SH,Dubey NK,Li WS,et al.Cordyceps militaris treatment preserves renal function in type 2 diabetic nephropathy mice[J].PLoS One,2016,11(11):e0166342.
[3]张亚莉.黄芪中主要化学成分的研究现状[J].临床合理用药,2011,4(2B):151-152.
[4]Guo Z,Xu HY,Xu L,et al.In vivo and in vitro immunomodulatory and anti-inflammatory and effects of total flavonoida of astragalus[J].Afr J Tradit Comp Altern Med,2016,13(4):60-73.
[5]Cottrell KA,Chaudhari HG,Cohen BA,et al.PTRE-seq reveals mechanism and interactions of RNA binding proteins and miRNAs[J].Nat Commun,2018,9(1):301.
[6]钟思思,辛柳燕,郑永亮,等.miR-378与肿瘤的研究进展[J].中国老年学,2015(24):7253-7256.
[7]邓海鸥,林凯,黎映兰,等.黄芪多糖对老年早期糖尿病肾病者TNF-α、IL-6和免疫功能的影响[J].中药材,2014,37(4):713-716.
[8]张体华,王明.糖尿病肾病患者炎症水平、免疫功能及与肾脏病变的关系[J].中国免疫学杂志,2016,32(10):1524-1526,1531.
[9]万莹,王羽,崔晓雪,等.黄芪总黄酮对柯萨奇B3病毒感染乳鼠心肌细胞miRNA378和miRNA378表达的影响[J].中国应用生理学杂志,2017,33(1):55-57.
[10]Zhang T,Hu J,Wang X,et al.MicroRNA-378 Promotes Hepatic Inflammation and Fibrosis via Modulation of the NFκB-TNFαPathway[J].J Hepatol,2019,70(1):87-96.
[11]陈建真,吕圭源,叶磊,等.黄芪黄铜的化学成分与药理作用研究进展[J].2009,28(10):1314-1316.
[12]Hei ZQ,Huang HQ,Zhang JJ,et al.Protective effect of Astragalus membranaceus on intestinal mucosa reperfusion injury after hemorrhagic shock in rats[J].World JGastroenterol,2005,11(32):4986-4991.
[13]Wang D,Zhuang Y,Tian Y,et al.Study of the effects of total flavonoids of Astragalus on atherosclerosis formation and potential mechanisms[J].Oxid Med Cell Longev,2012,2012:282383.
[14]郭泽,邹娟,张贤,等.黄芪总黄酮小鼠体内抗炎作用研究[J].动物医学进展,2015,36(9):64-66.
[15]戴国梁,朱立静,李长印,等.黄芪总黄酮对高血压大鼠肾脏TGF-β1/Smad信号通路的影响[J].药学与临床研究,2016,24(6):433-436.
[16]Liu S,Ye LF,Tao J,et al.Total flavones of Abelmoschus manihot improve diabetic nephropathy by inhibiting the iRhom2/TACE signalling pathway activity in rats[J].Pharm Biol,2017,56(1):1-11.
[17]Huang GM,Huang KY,Lee TY,et al.An interpretable rule-based diagnostic classification of diabetic nephropathy among type 2 diabetes patients[J].BMCBioinformatics,2015,16(1):S5.
[18]Mudaliar H,Pollock C,Panchapakesan U.Role of Toll-like receptors in diabetic nephropathy[J].Clin Sci(Lond),2014,126(10):685-694.
[19]Dhamodharan U,Viswanathan V,Krishnamoorthy E,et al.Genetic association of IL-6,TNF-αand SDF-1polymorphisms with serum cytokine levels in diabetic foot ulcer[J].Gene,2015,565(1):62-67.
[20]Arellano BAS,Tostado GM,Sánchez RO,et al.Immunomodulatory effects of the nutraceutical garlic derivative allicin in the progression of diabetic nephropathy[J].Int J Mol Sci,2018,19(10):3107.
[21]Li JX,Xu L,Sang R,et al.Immunomodulatory and antiinflammatory effects of total flavonoids of by regulating NF-κB and MAPK signalling pathways in RAW 264.7macrophages[J].Pharmazie,2018,73(10):589-593.
[2]Yu SH,Dubey NK,Li WS,et al.Cordyceps militaris treatment preserves renal function in type 2 diabetic nephropathy mice[J].PLoS One,2016,11(11):e0166342.
[3]张亚莉.黄芪中主要化学成分的研究现状[J].临床合理用药,2011,4(2B):151-152.
[4]Guo Z,Xu HY,Xu L,et al.In vivo and in vitro immunomodulatory and anti-inflammatory and effects of total flavonoida of astragalus[J].Afr J Tradit Comp Altern Med,2016,13(4):60-73.
[5]Cottrell KA,Chaudhari HG,Cohen BA,et al.PTRE-seq reveals mechanism and interactions of RNA binding proteins and miRNAs[J].Nat Commun,2018,9(1):301.
[6]钟思思,辛柳燕,郑永亮,等.miR-378与肿瘤的研究进展[J].中国老年学,2015(24):7253-7256.
[7]邓海鸥,林凯,黎映兰,等.黄芪多糖对老年早期糖尿病肾病者TNF-α、IL-6和免疫功能的影响[J].中药材,2014,37(4):713-716.
[8]张体华,王明.糖尿病肾病患者炎症水平、免疫功能及与肾脏病变的关系[J].中国免疫学杂志,2016,32(10):1524-1526,1531.
[9]万莹,王羽,崔晓雪,等.黄芪总黄酮对柯萨奇B3病毒感染乳鼠心肌细胞miRNA378和miRNA378表达的影响[J].中国应用生理学杂志,2017,33(1):55-57.
[10]Zhang T,Hu J,Wang X,et al.MicroRNA-378 Promotes Hepatic Inflammation and Fibrosis via Modulation of the NFκB-TNFαPathway[J].J Hepatol,2019,70(1):87-96.
[11]陈建真,吕圭源,叶磊,等.黄芪黄铜的化学成分与药理作用研究进展[J].2009,28(10):1314-1316.
[12]Hei ZQ,Huang HQ,Zhang JJ,et al.Protective effect of Astragalus membranaceus on intestinal mucosa reperfusion injury after hemorrhagic shock in rats[J].World JGastroenterol,2005,11(32):4986-4991.
[13]Wang D,Zhuang Y,Tian Y,et al.Study of the effects of total flavonoids of Astragalus on atherosclerosis formation and potential mechanisms[J].Oxid Med Cell Longev,2012,2012:282383.
[14]郭泽,邹娟,张贤,等.黄芪总黄酮小鼠体内抗炎作用研究[J].动物医学进展,2015,36(9):64-66.
[15]戴国梁,朱立静,李长印,等.黄芪总黄酮对高血压大鼠肾脏TGF-β1/Smad信号通路的影响[J].药学与临床研究,2016,24(6):433-436.
[16]Liu S,Ye LF,Tao J,et al.Total flavones of Abelmoschus manihot improve diabetic nephropathy by inhibiting the iRhom2/TACE signalling pathway activity in rats[J].Pharm Biol,2017,56(1):1-11.
[17]Huang GM,Huang KY,Lee TY,et al.An interpretable rule-based diagnostic classification of diabetic nephropathy among type 2 diabetes patients[J].BMCBioinformatics,2015,16(1):S5.
[18]Mudaliar H,Pollock C,Panchapakesan U.Role of Toll-like receptors in diabetic nephropathy[J].Clin Sci(Lond),2014,126(10):685-694.
[19]Dhamodharan U,Viswanathan V,Krishnamoorthy E,et al.Genetic association of IL-6,TNF-αand SDF-1polymorphisms with serum cytokine levels in diabetic foot ulcer[J].Gene,2015,565(1):62-67.
[20]Arellano BAS,Tostado GM,Sánchez RO,et al.Immunomodulatory effects of the nutraceutical garlic derivative allicin in the progression of diabetic nephropathy[J].Int J Mol Sci,2018,19(10):3107.
[21]Li JX,Xu L,Sang R,et al.Immunomodulatory and antiinflammatory effects of total flavonoids of by regulating NF-κB and MAPK signalling pathways in RAW 264.7macrophages[J].Pharmazie,2018,73(10):589-593.