神经药理学报

神经药理学报 ›› 2024, Vol. 14 ›› Issue (2): 7-.DOI: 10.3969/j.issn.2095-1396.2024.02.002

• 研究论文 • 上一篇    下一篇

交泰丸含药血清通过激活SIRT1 抑制内质网应激改善高糖高脂对SH-SY5Y 细胞的损伤

黎沛森   

  1. 广州广钢新城医院，广州，510145，中国
  • 出版日期:2024-04-26 发布日期:2024-07-10
  • 作者简介:黎沛森，主治医师；研究方向：中医药防治内科疾病；E-mail: 646293918@qq.com
  • 基金资助:
    国家自然科学基金项目（No.82004246）

Jiaotai Pill D rug-Containing Serum Ameliorates the Injury of SH-SY5Y Cells Induced by High Glucose/Palmitic Acid by Activating SIRT1 and Inhibiting Endoplasmic Reticulum Stress

LI Pei-sen   

  1. Guangzhou Guanggang Xincheng Hospital, Guangzhou, 510145, China
  • Online:2024-04-26 Published:2024-07-10

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摘要：

目的：探讨交泰丸改善高糖高脂对SH-SY5Y细胞损伤的可能作用机制。方法：将20只SD大鼠随机分 为交泰丸组和空白组，每组10只。交泰丸组予交泰丸药液8.4 g·kg-1灌胃，空白组予生理盐水灌胃，连续灌胃7 d， 制备交泰丸血清和空白血清。体外培养SH-SY5Y细胞，用不同比例血清给药处理24 h后，采用CCK-8法检测交 泰丸含药血清对SH-SY5Y细胞增殖活力影响；用25 mmol·L-1高糖和200 μmol·L-1棕榈酸造模，按含药血清比 例分为对照组和交泰丸含药血清低、高剂量组，SIRT1抑制剂EX527组。采用Western blot法检测各组细胞中内 质网相关蛋白，肌醇需酶1α(inositol-requiring enzyme 1α, IRE1α)、磷酸化-肌醇需酶1α(phospho-inositolrequiring enzyme 1α, p-IRE1α)、蛋白激酶R样内质网激酶(protein kinase R-like endoplasmic reticulum kinase, PERK)、磷酸化-蛋白激酶R样内质网激酶(phospho-protein kinase R-like endoplasmic reticulum kinase, p-PERK)、 激活性转录因子6(activating transcription factor 6, ATF6)、葡萄糖调节蛋白78(glucose regulated protein 78, GRP78)、蛋白二硫键异构酶(protein disulfide isomerase, PDI)及沉默信息调节因子1(silent information regulator 1, SIRT1)蛋白表达水平， 免疫荧光检测各组细胞中关键蛋白(SIRT1、p-IRE1α、p-PERK)的表达情况。 结果： 与对照组相比，模型组GRP78、p-PERK与PERK比值、p-IRE1α与IRE1α比值显著升高（P<0.05）,SIRT1、PDI 显著降低（P<0.05）；与模型组相比，交泰丸低、高浓度组SIRT1、PDI表达上调、GRP78、p-PERK与PERK比值、 p-IRE1α与IRE1α比值表达下调，且呈浓度依赖性（P<0.05），EX527组显著逆转交泰丸含药血清作用。结论：交 泰丸含药血清可改善高糖高脂对SH-SY5Y细胞的损伤，其作用机制可能与提高SIRT1表达，抑制内质网应激有 关。

关键词: 交泰丸, 高糖高脂, SIRT1, 内质网应激, 未折叠蛋白反应

Abstract:

Objective： To investigate the effect of Jiaotai Pill on SH-SY5Y cell injury induced by high glucose and high lipid. Methods： Twenty Sprague Dawley rats were randomly divided into control group and Jiaotai Pill group, with 10 rats in each group. The rats in the Jiaotai Pill group were given Jiaotai Pill solution 8.4 g·kg-1 by gavage, and the rats in the control group were given normal saline by gavage for 7 consecutive days to prepare blank serum and Jiaotai Pill serum. SH-SY5Y cells were cultured in vitro and treated with different proportions of serum for 24 hours. CCK-8 assay was used to detect the effect of serum containing Jiaotai Pill on the proliferation of SH-SY5Y cells. The cells were and divided into control group, low dose group and high dose group, SIRT1 inhibitor EX527 group according to the proportion of drug-containing serum and cultured with 25 mmol·L-1 high glucose and 200 μmol·L-1 palmitic acid or normal medium. Endoplasmic reticulum (ER) associated proteins [inositol-requiring enzyme 1α(IRE1α), phospho-inositol-requiring enzyme 1α(p-IRE1α), protein kinase R-like endoplasmic reticulum kinase (PERK), phospho-protein kinase R-like endoplasmic reticulum kinase (p-PERK), activating transcription factor 6(ATF6) and glucose regulated protein 78 (GRP78)、protein disulfide isomerase（PDI）] were detected by Western blot. Immunofluorescence was used to detect the expression of energy metabolism-related proteins (SIRT1、p-IRE1α、 p-PERK) in each group. Results： Compared with the control group, GRP78, the ratio of p-PERK to PERK and the ratio of p-IRE1α to IRE1α were significantly increased in the model group (P<0.05), SIRT1 and PDI were significantly decreased (P<0.05). Compared with the model group, the expressions of SIRT1 and PDI were up-regulated (P<0.05), and the expressions of GRP78, p-PERK/PERK,and p-IRE1α/IRE1α were down-regulated (P<0.05) in a concentrationdependent way in Jiaotai Pill low and high concentration groups.EX527 group significantly reversed the effect of Jiaotai pill(P<0.05). Conclusion： The drug-containing serum of Jiaotai Pill improve the high glucose/palmitic acid induced injury of SH-SY5Y cells by improving SIRT1 and inhibiting endoplasmic reticulum stress.

Key words: Jiaotai Pill, high glucose/palmitic acid, SIRT1, endoplasmic reticulum stress, unfolded protein response

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