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姚小磊,时健,刘倩宏,陈立浩,侯念婷.青光安Ⅱ号方对诱导损伤的RGC-5细胞中NF-κB/HIF-1α通路相关细胞因子的影响[J].湖南中医药大学学报英文版,2021,41(7):992-997.[Click to copy
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| 青光安Ⅱ号方对诱导损伤的RGC-5细胞中NF-κB/HIF-1α通路相关细胞因子的影响 |
| 姚小磊,时健,刘倩宏,陈立浩,侯念婷 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007;中医药防治眼耳鼻咽喉疾病湖南省重点实验室, 湖南 长沙 410208;中医药防治眼耳鼻咽喉疾病湖南省重点实验室, 湖南 长沙 410208;湖南中医药大学, 湖南 长沙 410208;广西骨伤医院, 广西 南宁 530001) |
| 摘要: |
| 目的 观察青光安Ⅱ号方对谷氨酸诱导损伤的RGC-5细胞中核因子κB（nuclear factor kappa-B，NF-κB）、低氧诱导因子-1α（hypoxia inducible factor-1，HIF-1α）、BCL2/腺病毒E1B相互作用蛋白3（BCL2/adenovirus E1B 19kDa interacting protein 3，BNIP3）、超氧化物歧化酶（superoxide dismutase，SOD）及丙二醛（malondialdehyde，MDA）的影响。方法 体外培养RGC-5细胞，分为4组：空白组、模型组、含药血清组和阻断剂组。模型组、含药血清组和阻断剂组予以谷氨酸诱导细胞损伤，模拟青光眼对神经节细胞的损伤，含药血清组加入青光安Ⅱ号方含药血清，阻断剂组加入KC7F2进行HIF-1α通路的阻断。以CCK-8法摸索含药血浆以及谷氨酸的最佳干预浓度；采用Hoechst法检测各组细胞的凋亡情况；Western blot检测NF-κB、HIF-1α、BNIP3蛋白表达情况；比色法检测SOD、MDA的表达情况。结果 CCK8法确定青光安Ⅱ号方5倍组含药血清为实验量，确定谷氨酸的最佳干预浓度为200 μM。与空白组比较，模型组NF-κB、HIF-1α、BNIP3、SOD、MDA表达显著升高（P<0.05）；与模型组比较，含药血清组、阻断剂组NF-κB、HIF-1α、BNIP3和SOD的表达显著降低（P<0.05）；含药血清组与阻断剂组的NF-κB、HIF-1α、BNIP3、SOD及MDA差异均无统计学意义（P>0.05）。结论 青光安Ⅱ号方对NF-κB具有抑制作用，进而抑制了HIF-1α相关通路的激活，减弱了由于氧化应激所致RGC-5细胞的凋亡，对RGC具有保护作用。 |
| 关键词: 青光眼 青光安Ⅱ号方 视网膜神经节细胞 核因子κB 低氧诱导因子-1α BCL2/腺病毒E1B相互作用蛋白3 超氧化物歧化酶 丙二醛 |
| DOI：10.3969/j.issn.1674-070X.2021.07.004 |
| Received:March 10, 2021 |
| 基金项目:国家自然科学基金地区基金项目（81860870）；湖南省自然科学基金项目（2018JJ3389）；第64批中国博士后科学基金面上资助一等资助项目（2018M640754）；中医药防治眼耳鼻咽喉疾病湖南省重点实验室建设项目（2017TP1018）；湖南中医药大学研究生创新课题项目（219CX69）。 |
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| Effects of Qingguangan Ⅱ Formula on NF-κB/HIF-1α Pathway Related Cytokines in Injury-induced RGC-5 Cells |
| YAO Xiaolei,SHI Jian,LIU Qianhong,CHEN Lihao,HOU Nianting |
| (The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Key Laboratory of Hunan Province for Prevention and Treatment of Eye, Ear, Nose and Throat Diseases with Traditional Chinese Medicine, Changsha, Hunan 410208, China;Key Laboratory of Hunan Province for Prevention and Treatment of Eye, Ear, Nose and Throat Diseases with Traditional Chinese Medicine, Changsha, Hunan 410208, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Guangxi Orthopedic Hospital, Nanning, Guangxi 530001, China) |
| Abstract: |
| Objective To observe the effects of Qingguangan Ⅱ Formula on nuclear factor kappa-B (NF-κB), hypoxia inducible factor-1 (HIF-1α), BCL2/adenovirus E1B 19kDa interacting protein 3 (BNIP3), superoxide dismutase (SOD) and malondialdehyde (MDA) in RGC-5 cells injury induced by glutamate. Methods The RGC-5 cells cultured in vitro were divided into 4 groups:blank group, model group, drug-containing serum group and blocker group. Glutamic acid was added to model group, drug-containing serum group and blocker group to simulate retinal ganglion cell damage caused by glaucoma, and Qingguangan Ⅱ Formula serum was added to drug-containing serum group, and the blocker group was added with KC7F2 to block the HIF-1α pathway. The CCK-8 method was used to find optimal intervention concentration of drug-containing plasma and glutamate; Hoechst method was used to detect the apoptosis of cells in each group; Western blot was used to detect the expression of NF-κB, HIF-1α, BNIP3; colorimetry was used to detect the expression of SOD and MDA. Results CCK8 method determined 5 times serum content as the Qingguangan Ⅱ Formula experimental amount, and the optimal intervention concentration of glutamate was 200 μM. Compared with blank group, the expression of NF-κB, HIF-1α, BNIP3, SOD and MDA increased significantly in the model group (P<0.05); compared with the model group, the expression of NF-κB, HIF-1α, BNIP3 and SOD in the drug-containing serum group and blocker group significantly decreased (P<0.05); there was no statistically significant difference between NF-κB, HIF-1α, BNIP3, SOD and MDA in drug-containing serum group and blocker group (P>0.05). Conclusion Qingguangan II Formula has an inhibitory effect on NF-κB, thereby inhibiting the activation of HIF-1α related pathways, reducing the apoptosis of RGC-5 cells caused by oxidative stress, it also has a protective effect on RGC. |
| Key words: Qingguangan Ⅱ Formula retinal ganglion cell nuclear factor kappa-B hypoxia inducible factor-1 BCL2/adenovirus E1B 19 kDa interacting protein 3 superoxide dismutase malondialdehyde |
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