王 珏,苏丽萍,严 岩,李雪会,张 婕,韩 静.甲基沿阶草酮甲对干眼症小鼠的治疗作用及机制[J].,2023,(23):4420-4426 |
甲基沿阶草酮甲对干眼症小鼠的治疗作用及机制 |
Therapeutic Effect and Mechanism of Methyl Ophiopogonanone A on Dry Eye Disease in Mice |
投稿时间:2023-03-07 修订日期:2023-04-03 |
DOI:10.13241/j.cnki.pmb.2023.23.004 |
中文关键词: 干眼症 甲基沿阶草酮甲 杯状细胞 氧化应激 炎症 |
英文关键词: Dry eye disease Methyl ophiopogonanone A Goblet cells Oxidative stress Inflammation |
基金项目:陕西省科技攻关重点研发计划项目(2021SF-158);唐都医院科技创新发展基金资助项目(2019LCYJ014) |
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中文摘要: |
摘要 目的:探讨甲基沿阶草酮甲(Methyl ophiopogonanone A,MOA)对干眼症(Dry eye disease,DED)小鼠的治疗作用及机制。方法:将雄性BALB/c小鼠(7~8周龄,体重18-22 g)随机分为5组(n=12):对照组、DED组、DED+10MOA组、DED+20MOA组和DED+40MOA组。对照组为正常小鼠,其他组小鼠双眼滴入质量浓度为0.2%的苯扎氯铵溶液诱导DED模型小鼠,每日1次,连续6周。建模后,对照组和DED组小鼠腹腔注射0.5 mL的1%二甲基亚砜溶液,DED+10MOA组、DED+20MOA组和DED+40MOA组小鼠依次腹腔注射0.5 mL剂量为10、20和40 mg/kg的MOA,每天1次,共给药28天。治疗结束后,检测各组小鼠的泪液分泌量和角膜荧光素钠染色分级,并进行角膜苏木素伊红(Hematoxylin and eosin,HE)染色、结膜过碘酸-雪夫(Periodic acid-Schiff,PAS)染色和角膜TUNEL染色。通过Western blot检测角膜K10蛋白表达水平。检测角膜氧化应激指标丙二醛(Malondialdehyde,MDA)和超氧化物歧化酶(Superoxide dismutase,SOD)水平。通过实时荧光定量逆转录聚合酶链式反应检测角膜中肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、白细胞介素-1β(Interleukin-1β,IL-1β)、白细胞介素-6(Interleukin-6,IL-6)、B淋巴细胞瘤-2相关X蛋白(Bcl2-associated X,Bax)和B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)的mRNA水平。结果:与对照组比较,DED组小鼠的泪液分泌量降低(P<0.05),角膜荧光素钠染色分级升高(P<0.05),角膜出现明显病变,结膜杯状细胞数量降低(P<0.05)。角膜Bcl-2 mRNA相对表达量、SOD水平降低(P<0.05),角膜TUNEL阳性率、Bax mRNA相对表达量、K10蛋白相对表达量、MDA、TNF-α、IL-1β和IL-6水平升高(P<0.05)。与DED组比较,DED+10MOA组、DED+20MOA组和DED+40MOA组的泪液分泌量升高(P<0.05),角膜荧光素钠染色分级降低(P<0.05),角膜病变减轻,结膜杯状细胞数量升高(P<0.05)。角膜TUNEL阳性率、Bax mRNA、K10蛋白相对表达量、MDA、TNF-α、IL-1β和IL-6水平降低(P<0.05),Bcl-2 mRNA相对表达量、SOD水平升高(P<0.05)。结论:甲基沿阶草酮甲有效减轻干眼症小鼠的症状及眼表病变,其机制可能与抑制杯状细胞凋亡和氧化应激及炎症反应有关。 |
英文摘要: |
ABSTRACT Objective: To explore the therapeutic effect and mechanism of methyl ophiopogonanone A (MOA) on dry eye disease (DED) rats. Methods: Male BALB/c mice (7~8 weeks old, weight 18-22 g) were randomly divided into 5 groups (n=12): control group, DED group, DED+10MOA group, DED+20MOA group and DED+40MOA group. Except the mice in control group, mice in other groups were given benzalkonium chloride solution with a mass concentration of 0.2% in both eyes to induce DED model mice, once a day for 6 weeks. After modeling, mice in the control group and DED group were intraperitoneally injected with 0.5 mL of 1% dimethyl sulfoxide solution, and mice in DED+10MOA group, DED+20MOA group and DED+40MOA group were intraperitoneally injected with 0.5 mL 10, 20 and 40 mg/kg/d MOA, once a day for 28 days. After treatment, the tear secretion and corneal fluorescein sodium staining grade of mice in each group were detected, and corneal hematoxylin and eosin (HE) staining, conjunctival periodic acid-Schiff (PAS) staining and corneal TUNEL staining were performed. The expression level of corneal K10 protein was detected by Western blot. The levels of corneal oxidative stress indicators malondialdehyde (MDA) and superoxide dismutase (SOD) were detected. The mRNA levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), Bcl2-associated X (Bax) and B-cell lymphoma-2 (Bcl-2) in cornea were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction. Results: Compared with control group, the tear secretion in DED group decreased (P<0.05), the grading of corneal fluorescein sodium staining increased (P<0.05), corneal lesions appeared, the number of conjunctival goblet cells decreased (P<0.05), corneal Bcl-2 mRNA relative expression level and SOD level decreased (P<0.05), corneal TUNEL positive rate, Bax mRNA relative expression level, K10 protein relative expression level, MDA, TNF-α, IL-1β and IL-6 levels increased (P<0.05). Compared with DED group, the tear secretion in DED+10MOA group, DED+20MOA group and DED+40MOA group increased (P<0.05), the grading of corneal fluorescein sodium staining decreased (P<0.05), the corneal lesions were alleviated, the number of conjunctival goblet cells increased (P<0.05). The positive rate of TUNEL, Bax mRNA, relative expression level of K10 protein, MDA, TNF-α, IL-1β and IL-6 levels decreased (P<0.05), while the relative expression level of Bcl-2 mRNA and SOD level increased (P<0.05). Conclusion: Methylophiopogonanone A effectively alleviates the symptoms and ocular surface lesions in mice with dry eye disease, and its mechanism may be related to the inhibition of goblet cell apoptosis, oxidative stress, and inflammation. |
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