秦莉, 陈宝光, 郭丰年, 张秋菊. 竹节参皂苷Ⅳa调控环状RNA EIF4G2/微小RNA-379-5p对高糖诱导的人肾小管上皮细胞损伤的影响[J]. 临床肾脏病杂志, 2022, 22(6): 492-497. DOI: 10.3969/j.issn.1671-2390.2022.06.008
    引用本文: 秦莉, 陈宝光, 郭丰年, 张秋菊. 竹节参皂苷Ⅳa调控环状RNA EIF4G2/微小RNA-379-5p对高糖诱导的人肾小管上皮细胞损伤的影响[J]. 临床肾脏病杂志, 2022, 22(6): 492-497. DOI: 10.3969/j.issn.1671-2390.2022.06.008
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    Qin Li, Chen Bao-guang, Guo Feng-nian, Zhang Qiu-ju. Effects of chikusetsu saponin Ⅳa on high glucose-induced human renal tubular epithelial cell injury through regulating circEIF4G2/miR-379-5p[J]. Journal of Clinical Nephrology, 2022, 22(6): 492-497. DOI: 10.3969/j.issn.1671-2390.2022.06.008
    Citation: Qin Li, Chen Bao-guang, Guo Feng-nian, Zhang Qiu-ju. Effects of chikusetsu saponin Ⅳa on high glucose-induced human renal tubular epithelial cell injury through regulating circEIF4G2/miR-379-5p[J]. Journal of Clinical Nephrology, 2022, 22(6): 492-497. DOI: 10.3969/j.issn.1671-2390.2022.06.008
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    竹节参皂苷Ⅳa调控环状RNA EIF4G2/微小RNA-379-5p对高糖诱导的人肾小管上皮细胞损伤的影响

    Effects of chikusetsu saponin Ⅳa on high glucose-induced human renal tubular epithelial cell injury through regulating circEIF4G2/miR-379-5p

      摘要
    • 摘要: 目的 探讨竹节参皂苷Ⅳa（chikusetsu saponin Ⅳa,CHS）对高糖诱导的人肾小管上皮细胞（human renal tubular epithelial cells,HK-2）损伤的影响,探讨其机制是否与调控环状RNA EIF4G2（circular RNA EIF4G2,circEIF4G2）和微小RNA-379-5p（microRNA-379-5p,miR-379-5p）表达有关。方法 HK-2细胞分为正常糖（normal glucose,NG）组（5.0 mmol/L葡萄糖）、高糖（high glucose,HG）组、高渗（mannitol glucose,MG）组、HG+CHS 25 μM组、HG+CHS 50 μM组、HG+CHS 100 μM组、HG+si-NC组、HG+si-circEIF4G2组、HG+CHS+pcDNA组、HG+CHS+pcDNA-circEIF4G2组。流式细胞术检测细胞凋亡率,试剂盒分析细胞内丙二醛（malondialedhyde,MDA）水平和超氧化物歧化酶（superoxide dismutase,SOD）活性。RT-qPCR测定circEIF4G2和miR-379-5p水平。双荧光素酶报告实验验证circEIF4G2和miR-379-5p靶向关系。结果 与NG组比较,HG组HK-2细胞凋亡率（7.62±0.74）%比（23.81±1.45）%、MDA水平（139.17±13） nmol/mL比（637.50±59.93）nmol/mL、circEIF4G2相对水平（0.96±0.07）比（3.00±0.15）显著升高（P<0.05）,SOD活性（165.05±16.52）U/L比（55.77±7.30）U/L、miR-379-5p相对水平（1.00±0.00）比（0.14±0.02）显著降低（P<0.05）。与HG组比较,HG+CHS 25 μM组、HG+CHS 50 μM组、HG+CHS 100 μM组HK-2细胞凋亡率（23.81±1.45）%比（20.70±1.07）%比（16.78±0.87）%比（11.06±0.77）%、MDA水平（637.50±59.93） nmol/mL比（533.36±17.83） nmol/mL比（407.51±34.34） nmol/mL比（276.04±15.15）nmol/mL、circEIF4G2相对水平（3.00±0.15）比（2.26±0.10）比（1.75±0.07）比（1.33±0.11）显著降低（P<0.05）,SOD活性（55.77±7.30）U/L比（79.99±8.95）U/L比（118.67±11.73）U/L比（144.78±8.52）U/L、miR-379-5p相对水平（0.14±0.02）比（0.29±0.03）比（0.48±0.06）比（0.75±0.07）显著升高（P<0.05）。circEIF4G2直接靶向结合miR-379-5p。与HG+si-NC组比较,HG+si-circEIF4G2组HK2细胞凋亡率（23.93±1.32）%比（13.77±1.18）%、MDA水平（642.24±52.33） nmol/mL比（311.91±20.40）nmol/mL显著降低（P<0.05）,SOD活性（55.41±7.75）U/L比（130.67±13.17）U/L、miR-379-5p相对水平（1.00±0.00）比（4.13±0.11）显著升高（P<0.05）。与HG+CHS+pcDNA组比较,HG+CHS+pcDNA-circEIF4G2组HK2细胞凋亡率（11.14±0.83）%比（19.74±1.14）%、MDA水平（277.89±15.46）nmol/mL比（583.39±10.29）nmol/mL显著升高（P<0.05）,SOD活性（144.04±13.23）U/L比（62.98±6.14）U/L显著降低（P<0.05）。结论 CHS可减轻高糖诱导的HK-2凋亡和氧化应激损伤,其机制与抑制circEIF4G2/miR-379-5p轴有关。

       

      Abstract: Objective To explore the effect of chikusetsu saponin Ⅳa (CHS) on high glucoseinduced human renal tubular epithelial cells (HK-2) injury and examine whether or not its mechanism is correlated with the regulation of circEIF4G2 and miR-379-5p expression. Methods HK-2 cells were divided into 10 groups of normal glucose (NG) (5.0 mmol/L glucose),high glucose (HG),HG+CHS 25 μM,HG+CHS 50 μM,HG+CHS 100 μM,HG+si-NC,HG+si-circEIF4G2,HG+CHS+ pcDNA and HG+CHS+pcDNA-circEIF4G2. Flow cytometry was performed for detecting apoptotic rate and kit employed for evaluating intracellular malondialdehyde (MDA) level and superoxide dismutase (SOD) activity. The levels of circEIF4G2 and miR-379-5p were assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). And targeting relationship between circEIF4G2 and miR-379-5p was verified by dual lucifase reporting assay. Results Compared with NG group,apoptotic rate (7.62±0.74)% vs (23.81±1.45)%,MDA level (139.17±13.60) nmol/mL vs (637.50±59.93) nmol/mL and relative level of circEIF4G2 (0.96±0.07) vs (3.00±0.15) spiked markedly in HG group (P<0.05) while SOD activity (165.05±16.52) U/L vs (55.77±7.30) U/L and relative level of miR-379-5p (1.00±0.00) vs (0.14±0.02) dropped obviously (P<0.05). Compared with HG group,apoptotic rate (23.81±1.45)% vs (20.70±1.07)% vs (16.78±0.87)% vs (11.06±0.77)%,MDA level (637.50±59.93) nmol/mL vs (533.36±17.83) nmol/mL vs (407.51±34.34) vs (276.04±15.15) nmol/mL and relative level of circEIF4G2 (3.00±0.15) vs (2.26±0.10) vs (1.75±0.07) vs (1.33±0.11) declined markedly in HG+CHS 25/50/100 μM group (P<0.05) while SOD activity (55.77±7.30) U/L vs (79.99±8.95) U/L vs (118.67±11.73) U/L vs (144.78±8.52) U/L and relative level of miR-379-5p (0.14±0.02) vs (0.29±0.03) vs (0.48±0.06) vs (0.75±0.07) spiked obviously (P<0.05). And circEIF4G2 was directly conjugated with miR-379-5p. Compared with HG+si-NC group,apoptotic rate (23.93±1.32)% vs (13.77±1.18)% and MDA level (642.24±52.33) nmol/mL vs (311.91±20.40) nmol/mL declined markedly in HG+si-circEIF4G2 group (P<0.05) and SOD activity (55.41±7.75) U/L vs (130.67±13.17) U/L and relative level of miR-379-5p rose significantly (1.00±0.00) vs (4.13±0.11) (P<0.05). Compared with HG+CHS+pcDNA group,apoptotic rate (11.14±0.83)% vs (19.74±1.14)% and MDA level (277.89±15.46) nmol/mL vs (583.39±10.29) nmol/mL spiked markedly in HG+CHS+pcDNA-circEIF4G2 group (P<0.05) while SOD activity dropped markedly (144.04±13.23) U/L vs (62.98±6.14) U/L,P<0.05. Conclusion Chikusetsu saponin Ⅳa may alleviate high glucose-induced human renal tubular epithelial cell apoptosis and oxidative stress injury. The mechanism is correlated with a suppression of circEIF4G2/miR-379-5p axis.

       

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