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小檗胺对糖尿病肾小管上皮细胞损伤的影响

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目的 研究小檗胺对体外糖尿病肾小管上皮细胞(RTEC)损伤的影响.方法 将RTEC分成对照组、模型组(高糖)、低剂量实验组(2 mg·L-1 小檗胺和高糖)、中剂量实验组(4 mg·L-1 小檗胺和高糖)、高剂量实验组(8 mg·L-1小檗胺和高糖)、BBM-H+miR-NC组(转染mimics control,8 mg·L-1小檗胺和高糖)、BBM-H+miR-135b组(转染miR-135b mimics,8 mg·L-1 小檗胺和高糖).以噻唑蓝(MTT)法检测细胞增殖活性,以流式细胞术检测细胞凋亡率,以蛋白质印迹法检测细胞B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X(Bax)蛋白表达,以化学荧光法检测活性氧(ROS)水平,以硫代巴比妥酸法检测丙二醛(MDA)水平,以黄嘌呤氧化法检测超氧化物歧化酶(SOD)水平,以酶联免疫吸附试验(ELISA)法检测肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)和白细胞介素-1β(IL-1β)水平.结果 对照组、模型组和低、中、高剂量实验组的 RTEC 增殖活性(OD 值)分别为 0.66±0.04、0.36±0.02、0.43±0.03、0.54±0.03 和 0.63±0.05,细胞 凋亡率 分别为(3.62±0.31)%、(29.41±2.33)%、(20.10±1.65)%、(15.02±1.25)%和(9.58±1.43)%,MDA 分别为(1.04±0.12)、(5.24±0.29)、(3.45±0.22)、(2.16±0.13)和(1.60±0.11)nmol·mL-1,SOD 分别为(240.22±12.06)、(130.56±10.84)、(169.62±12.50)、(201.97±12.78)和(236.74±14.52)U·mL-1,TNF-α 分别为(31.25±2.51)、(51.84±4.20)、(44.52±2.61)、(38.25±1.50)和(32.10±1.78)mg·L-1,IL-8 分别 为(10.59±1.14)、(19.95±1.74)、(16.10±1.03)、(13.52±1.25)和(11.17±0.92)mg·L-1,IL-1β 分别 为(23.01±1.45)、(56.92±2.51)、(43.20±1.96)、(32.05±1.23)和(26.37±2.48)mg·L-1.模型组的上述指标与对照组比较,在统计学上差异均有统计学意义(均P<0.05);低、中、高剂量实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P<0.05);BBM-H+miR-NC组与BBM-H+miR-135b组的上述指标比较,在统计学上差异均有统计学意义(均P<0.05).结论 小檗胺通过下调miR-135b减少体外糖尿病RTEC损伤.
Effects of berberine on renal tubular epithelial cell injury in diabetic
Objective To study the effect of berberine on the damage of renal tubular epithelial cells(RETC)in diabetic in vitro.Methods RTEC were divided into control group,model group(high glucose),low dose experimental group(2 mg·L-1 berberine and high glucose),medium dose experimental group(4 mg·L-1 berberine and high glucose),high dose experimental group(8 mg·L-1 berberine and high glucose),BBM-H+miR-NC(transfected with mimics control group,8 mg·L-1 berberine and high glucose),BBM-H+miR-135b group(transfected with miR-135b mimics,8 mg·L-1 berberine and high glucose).Methyl thiazolyl tetrazolium(MTT)method was used to detect cell proliferation activity,flow cytometry was used to detect cell apoptosis rate,Western blot was used to detect the expression of B-cell lymphoma-2(Bcl-2)and Bel-associated X(Bax)proteins in cells,and reactive oxygen species(ROS)level were detected by chemical fluorescence method,thiobarbituric acid method was used to detect malondialdehyde(MDA)level,and xanthine oxidation method was used to detect superoxide dismutase(SOD)level,the levels of tumor necrosis factor-α(TNF-α),interleukin-8(IL-8)and interleukin-1β(IL-1 β)were detected by enzyme-linked immunosorbent assay(ELISA).Results The proliferative activity(OD value)of RTEC in control group,model group and low,medium,high dose experimental groups were 0.66±0.04,0.36±0.02,0.43±0.03,0.54±0.03,0.63±0.05;the apoptosis rates were(3.62±0.31)%,(29.41±2.33)%,(20.10±1.65)%,(15.02±1.25)%,(9.58±1.43)%;MDA were(1.04±0.12),(5.24±0.29),(3.45±0.22),(2.16±0.13),(1.60±0.11)nmol·mL-1;SOD were(240.22±12.06),(130.56±10.84),(169.62±12.50),(201.97±12.78),(236.74±14.52)U·mL-1;TNF-α were(31.25±2.51),(51.84±4.20),(44.52±2.61),(38.25±1.50),(32.10±1.78)mg·L-1;IL-8 were(10.59±1.14),(19.95±1.74),(16.10±1.03),(13.52±1.25),(11.17±0.92)mg·L-1;IL-1β were(23.01±1.45),(56.92±2.51),(43.20±1.96),(32.05±1.23),(26.37±2.48)mg·L-1.There were statistically significant differences between model group and control group(all P<0.05).Compared with the model group,the above indexes in low,medium and high dose experimental group were statistically significant(all P<0.05).The above indexes of BBM-H+miR-NC group were statistically significant compared with those of BBM-H+miR-135b group(all P<0.05).Conclusion Berberine can reduce diabetic renal tubular epithelial cell damage by down-regulating miR-135b.

berberinediabetesrenal tubular epithelial cells

张鹏飞、布琼星、衡艳艳、王斐斐、魏巍

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长治医学院 附属和平医院 肾内科,山西长治 046000

小檗胺 糖尿病 肾小管上皮细胞

山西省基础研究计划基金资助项目

202303021211107

2024

中国临床药理学杂志
中国药学会

中国临床药理学杂志

CSTPCD北大核心
影响因子:1.91
ISSN:1001-6821
年,卷(期):2024.40(3)
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