姚小磊 时健 刘倩宏 陈立浩 侯念婷

〔摘要〕 目的 觀察青光安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

〔中图分类号〕R276.7       〔文献标志码〕A       〔文章编号〕doi：10.3969/j.issn.1674-070X.2021.07.004

〔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.

〔Keywords〕 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

青光眼為导致人类失明的三大眼类疾病之一。调查显示，2020年全球青光眼患病人数高达7 600万，其中我国占2 100万，位居世界之首[1]，目前，超过40岁以上人群青光眼的总患病康复率仅有1.5%～3.6%[2]，治疗方案主要以控制眼压来保存视力，但是青光眼导致的视神经损伤却不可逆，且没有较好的视神经保护方案。彭清华教授通过多年临床经验总结出“青光安Ⅱ号方”，研究表明其有较好的临床疗效，尤其对青光眼术后仍存在视力下降的患者，有较好的视神经保护效应[3]，但其具体的机制并不清楚。……