摘要
目的探讨益气养阴活血方通过转录共激活因子P300(P300)、沉默调节因子1(SIRT1)对高糖诱导的肾小管上皮细胞(NRK-52E细胞)代谢记忆的影响。方法将NRK-52E细胞按照低糖组、高糖组、短暂高糖组、高糖+中药血清治疗组、短暂高糖+中药血清治疗组进行分组干预后,免疫组化法检测细胞α-平滑肌激动蛋白(α-SMA)和E-钙黏蛋白(E-Cadherin)表达,RT-PCR法检测P300、SIRT1 mRNA表达。结果 (1)高糖组NRK-52E细胞α-SMA表达显著升高,E-Cadherin随着高糖刺激时间增加表达逐渐降低。短暂高糖组α-SMA表达较正常组仍有明显升高,E-Cadherin表达较正常组略微减少。中药血清干预后α-SMA表达减弱,高糖刺激时间越短,干预效果越明显,中药血清干预后E-Cadherin表达明显增加。(2)高糖刺激后SIRT1 mRNA表达相对降低,P300mRNA表达相对增加,与低糖组比较差异均有统计学意义(P<0.05)。高糖组与短暂高糖组比较,SIRT1 mRNA与P300 mRNA表达均未见明显差异(P>0.05)。高糖+中药血清治疗组与高糖组比较,SIRT1 mRNA表达相对增加,P300 mRNA表达相对降低,差异均有统计学意义(P<0.05)。短暂高糖+中药血清治疗组与短暂高糖组比较,SIRT1 mRNA表达相对增加,P300表达相对降低,差异均有统计学意义(P<0.05)。高糖+中药血清治疗组与短暂高糖+中药血清治疗组比较,SIRT1与P300 mRNA表达均未见明显差异(P>0.05)。结论益气养阴活血方可能抑制P300,同时刺激SIRT1表达,并通过组蛋白乙酰化修饰改变抑制炎症相关基因转录,减轻NRK-52E细胞凋亡及肾损伤,改善代谢记忆。
Objective To investigate the effects of Supplementing Qi, nourishing Yin and activating blood（SQNYAB） decoction on metabolic memory of renal tubular epithelial cell（NRK-52 E cells）cultured in high con centrations of glucose through transcription coactivators P300 and silent information regulator-1（SIRT1）. Methods NRK-52 E cells were divided into five groups： the low-concentration glucose group, the high-concentration glucose group, the short-time high-concentrations glucose group, the high-concentration glucose＋serum containing SQNYAB decoction group and the short-time high-concentrations glucose ＋ serum containing SQNYAB decoction group. The expressions of smooth muscle agonist protein-α（α-SMA） and E-cadherin were observed by immunohistochemical analysis. The mRNA expression of P300 and SIRT1 were analyzed by RT-PCR. Results（1） The expression ofα-SMA significantly elevated in the high-concentrations glucose group, while the expression of E-cadherin decreased with time. In the short-time high-concentrations glucose group, the expression of α-SMA also elevated obviously compare to the control group, while the expression of E-cadherin slightly decreased. The expression of α-SMA dropped down and the expression of E-cadherin went up after SQNYAB decoction treatment. This intervention effect enhanced with treatment time.（2） The mRNA expression of SIRT decreased, while the expression of P300 increased in high-concentrations glucose group, and showed statistical significance compare to the low-concentration glucose group（P〈0.05）. There was no significant differences between the short-time high-concentrations glucose group and the high-concentrations glucose group（P〉0.05）. The mRNA expression of SIRT increased, while the expression of P300 decreased in high-concentrations glucose＋serum containing SQNYAB decoction group, and showed statistical significance compare to the high-concentration glucose group（P〈0.05）. There was no significant differences between the short-t
作者
章文星
杜月光
柴可夫
ZHANG Wenxing;DU Yueguang;CHAI Kefu(School of Basic Medicine,Zhejiang Chinese Medical University,Hangzhou（310053）,China)
出处
《浙江中西医结合杂志》
2018年第7期533-536,613,共5页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
基金
国家自然科学基金(No.81774177)
浙江省自然科学基金(No.YL16H270008)
