[1]罗丽丹,李达佑,钟华,等.地黄多糖对过氧化氢诱导的人晶状体上皮细胞氧化损伤的保护作用[J].眼科新进展,2022,42(8):612-616.[doi:10.13389/j.cnki.rao.2022.0125]
　LUO Lidan,LI Dayou,ZHONG Hua,et al.Protective effect of Rehmannia glutinosa polysaccharide on hydrogen peroxide-induced oxidative damage in human lens epithelial cells[J].Recent Advances in Ophthalmology,2022,42(8):612-616.[doi:10.13389/j.cnki.rao.2022.0125]
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地黄多糖对过氧化氢诱导的人晶状体上皮细胞氧化损伤的保护作用/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
42卷
期数:
2022年8期
页码:
612-616
栏目:
实验研究
出版日期:
2022-08-06

文章信息/Info

Title:
Protective effect of Rehmannia glutinosa polysaccharide on hydrogen peroxide-induced oxidative damage in human lens epithelial cells
作者:
罗丽丹; 李达佑; 钟华; 曹玲英; 刘婷婷
514011　广东省梅州市，梅州市人民医院眼科（罗丽丹，李达佑，曹玲英，刘婷婷）；514011　广东省梅州市，梅州市人民医院产前诊断中心实验室（钟华）
Author(s):
LUO Lidan1;  LI Dayou1;  ZHONG Hua2;  CAO Lingying1;  LIU Tingting1
1.Department of Ophthalmology, Meizhou People’s Hospital, Meizhou 514011, Guangdong Province, China
2.Prenatal Diagnostic Center Laboratory, Meizhou People’s Hospital, Meizhou 514011, Guangdong Province, China
关键词:
地黄多糖; 过氧化氢; 人晶状体上皮细胞; NOD样受体蛋白3/ caspase-1通路; 细胞焦亡
Keywords:
Rehmannia glutinosa polysaccharide;  hydrogen peroxide;  human lens epithelial cells;  NOD-like receptor protein 3/caspase-1 pathway;  pyroptosis
分类号:
R776
DOI:
10.13389/j.cnki.rao.2022.0125
文献标志码:
A
摘要:
目的　探讨地黄多糖对过氧化氢（H2O2）诱导的人晶状体上皮细胞（HLEC）氧化损伤的保护作用及机制。方法　取对数生长期的B-3细胞（HLEC）作为研究对象。筛选H2O2和地黄多糖最佳作用浓度。依据筛选的地黄多糖抑制H2O2致B-3细胞损伤的最佳作用浓度处理细胞，将B-3细胞分为：空白组、H2O2组和地黄多糖组。空白组：用10 g·L-1羧甲基纤维素钠预处理细胞24 h后，换为EMEM培养基培养24 h；H2O2组：用10 g·L-1羧甲基纤维素钠预处理细胞24 h后，换为含100 mol·L-1 H2O2的EMEM培养基培养24 h；地黄多糖组：用含40 mg·L-1 地黄多糖的培养基预处理细胞24 h后，换为含100 mol·L-1 H2O2的EMEM培养基培养24 h。采用扫描电镜和免疫荧光染色观察细胞焦亡情况；测定细胞活性氧（ROS）水平、丙二醛（MDA）含量及培养液中白细胞介素（IL）-1β、IL-18水平；Western blot检测NOD样受体蛋白3（NLRP3）/ caspase-1途径相关蛋白表达水平。结果　100 mol·L-1 H2O2和40 mg·L-1 地黄多糖为最佳作用浓度。扫描电镜结果显示：空白组细胞呈圆球形，边界规则；H2O2组细胞肿胀变大，边界不规则；地黄多糖组细胞肿胀程度减轻。与H2O2组相比，地黄多糖组切割消皮素D（GSDMD）-N阳性细胞数为每视野（5.33±0.46）个，GSDMD-N阳性细胞数显著减少（P<0.05）。与H2O2组相比，地黄多糖组B-3细胞ROS水平、MDA含量显著降低（均为P<0.05）。与H2O2组相比，地黄多糖组培养液中IL-1β、IL-18含量降低（q=20.694、11.618，均为P<0.05）。与H2O2组相比，地黄多糖组B-3细胞NLRP3蛋白表达水平及caspase-1/pro-caspase-1、GSDMD-N/GSDMD均降低（均为P<0.05）。结论　地黄多糖可减轻H2O2作用下B-3细胞的氧化损伤，其作用机制与抑制细胞焦亡有关。
Abstract:
Objective　To investigate the protective effect and mechanism of Rehmannia glutinosa polysaccharide (RGP) on hydrogen peroxide (H2O2)-induced oxidative damage in human lens epithelial cells (HLEC). Methods　B-3 cells (HLEC) in the logarithmic growth stage were selected for this study. The optimal concentration of H2O2 and RGP were screened. Based on the optimal concentration of RGP to inhibit H2O2-induced damage in B-3 cells, B-3 cells were divided into the blank group, H2O2 group, and RGP group. Cells in the blank group were pretreated with 10 g·L-1 sodium carboxymethylcellulose for 24 h and then cultured in EMEM medium for 24 h. Cells in the H2O2 group were pretreated with 10 g·L-1 sodium carboxymethylcellulose for 24 h and then cultured in EMEM medium containing 100 mol·L-1 H2O2 for 24 h. Cells in the RGP group were pretreated with medium containing 40 mg·L-1 RGP for 24 h and then cultured in EMEM medium containing 100 mol·L-1 H2O2 for 24 h. Cell pyroptosis was assessed by scanning electron microscope and immunofluorescence. The levels of reactive oxygen species (ROS) and malondialdehyde (MDA) in cells and the levels of interleukin (IL)-1β and IL-18 in the culture medium were measured. The levels of NOD-like receptor protein 3 (NLRP3)/caspase-1 pathway related proteins were detected by Western blot. Results　The optimal concentration of H2O2 and RGP in the study was 100 mol·L-1 and 40 mg·L-1, respectively. The scanning electron microscope results showed that cells in the blank group were round and spherical with regular borders; cells in the H2O2 group became swollen with irregular borders; and cells in the RGP group were less swollen. The number of gasdermin-D (GSDMD)-N+ cells was (5.33±0.46) per field in the RGP group, which was significantly lower than that in the H2O2 group (P<0.05). Compared with the H2O2 group, the ROS and MDA levels in the RGP group were significantly reduced (both P<0.05), and so were the IL-1β and IL-18 levels (q=20.694 and 11.618, both P<0.05). Compared with the H2O2 group, the protein level of NLRP3 and the ratios of caspase-1/pro-caspase-1 and GSDMD-N/GSDMD in the RGP group decreased (all P<0.05). Conclusion　RGP can alleviate the H2O2-induced oxidative damage in B-3 cells by inhibiting pyroptosis.

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备注/Memo

备注/Memo:
梅州市医药卫生科研课题（编号：2015-B-22）
更新日期/Last Update: 2022-08-05