白蒺藜抗光损伤小鼠光感受器细胞凋亡及保护视网膜功能的研究
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摘要
目的探讨白蒺藜对光损伤小鼠光感受器细胞凋亡及受损视网膜功能的影响。方法 BALB/c小鼠随机分为正常对照组、光损伤模型组、白蒺藜给药组。暗适应24 h后,白蒺藜给药组腹腔注射白蒺藜水煎液100μl(每20 g体质量100 mg生药),给药后30 min进行光照造模。造模采用白色荧光灯,光强度为10 000 lux,持续照射30 min。正常对照组和光损伤模型组同时腹腔注射等量0.9%NaCl溶液。(1)光照1 d、3 d后,摘取眼球制备冰冻切片,采用TUNEL法标记凋亡的光感受器细胞。(2)光照6 d后采用视网膜电流图(ERG)检测小鼠视网膜功能。结果 (1)TUNEL结果显示,正常对照组的外核层光感受器细胞核TUNEL染色为阴性。与正常对照组比较,光损伤后模型组小鼠的视网膜外核层TUNEL阳性光感受器在1 d开始显著增加,3 d达到高峰(P<0.001);与光损伤模型组比较,白蒺藜给药组外核层仅见少量散在的TUNEL阳性光感受器细胞(P<0.05)。(2)ERG结果显示,与正常对照组比较,光损伤模型组小鼠视网膜a波、b波振幅显著降低(P<0.05),白蒺藜给药组a波、b波振幅显著高于模型组(P<0.05)。结论白蒺藜能显著地减轻光损伤引起的光感受器细胞凋亡,对光损伤小鼠视网膜功能具有保护作用。
Objective To investigate the effect of Fructus Tribulus( TTL) on photoreceptor cell apoptosis and impaired retina in the mouse model of bright light-induced retinal degeneration. Methods BALB/c mice were randomly divided into normal control group,bright light-exposed group and TTL-treated group. After dark adaptation for 24 h,bright light was applied at the intensity of 10 000 Lux for 30 min. Totally 100 μl of TTL decoction( 100 mg per 20 g body mass) was intraperitoneally administered 30 min prior to bright light exposure in the TTL-treated group. The normal control group and the bright light-exposed group were intraperitoneally injected with the same amount of 0.9%NaCl solution.(1)Eyes were enucleated 1 d and 3 d later and subjected to frozen section for the detection of apoptotic cells using TUNEL assay.(2)The retinal function of mice was measured by electroretinogram( ERG) 6 d after bright light exposure. Results(1)In the normal control group,TUNEL staining was negative in the photoreceptor cell nucleus of outer nuclear layer. Compared with the normal control group,the TUNEL positive photoreceptor cells in the outer nuclear layer of retina increased significantly on the 1 st day and reached the peak at the 3 rd day after light injury in bright light-exposed group( P<0.001). Compared with the bright light-exposed group,only a few scattered TUNEL positive photoreceptor cells were found in the outer nuclear layer in the TTL-treated group( P<0.05).(2)ERG revealedthat the amplitudes of a-wave and b-wave were significantly depressed in the bright light-exposed group compared with those in the normal control group( P < 0.05). In contrast,the amplitudes of a-wave and b-wave in TTL-treated group were significantly higher than those in the bright light-exposed group( P < 0. 05). Conclusions Tribulus terrestris L. can significantly reduce photoreceptor cell apoptosis induced by light injury,and has protective effect on retinal function in mice.
Objective To investigate the effect of Fructus Tribulus( TTL) on photoreceptor cell apoptosis and impaired retina in the mouse model of bright light-induced retinal degeneration. Methods BALB/c mice were randomly divided into normal control group,bright light-exposed group and TTL-treated group. After dark adaptation for 24 h,bright light was applied at the intensity of 10 000 Lux for 30 min. Totally 100 μl of TTL decoction( 100 mg per 20 g body mass) was intraperitoneally administered 30 min prior to bright light exposure in the TTL-treated group. The normal control group and the bright light-exposed group were intraperitoneally injected with the same amount of 0.9%NaCl solution.(1)Eyes were enucleated 1 d and 3 d later and subjected to frozen section for the detection of apoptotic cells using TUNEL assay.(2)The retinal function of mice was measured by electroretinogram( ERG) 6 d after bright light exposure. Results(1)In the normal control group,TUNEL staining was negative in the photoreceptor cell nucleus of outer nuclear layer. Compared with the normal control group,the TUNEL positive photoreceptor cells in the outer nuclear layer of retina increased significantly on the 1 st day and reached the peak at the 3 rd day after light injury in bright light-exposed group( P<0.001). Compared with the bright light-exposed group,only a few scattered TUNEL positive photoreceptor cells were found in the outer nuclear layer in the TTL-treated group( P<0.05).(2)ERG revealedthat the amplitudes of a-wave and b-wave were significantly depressed in the bright light-exposed group compared with those in the normal control group( P < 0.05). In contrast,the amplitudes of a-wave and b-wave in TTL-treated group were significantly higher than those in the bright light-exposed group( P < 0. 05). Conclusions Tribulus terrestris L. can significantly reduce photoreceptor cell apoptosis induced by light injury,and has protective effect on retinal function in mice.
引文
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[2] TANG L,BAO S,DU Y,et al.Antioxidant effects of Lycium barbarum polysaccharides on photoreceptor degeneration in the light-exposed mouse retina[J].Biomed Pharmacother,2018(103):829-837.
[3]谢妍,沈澍农.浅析张简斋治疗眼病的辨治经验[J].中国民间疗法,2018,26(9):51-52.
[4]王倩,刘子豪.白蒺藜的临床应用研究进展[J].中西医结合心脑血管病杂志,2016,14(16):1877-1879.
[5]吴晗晗,徐静,卞敏娟,等.石斛夜光丸光感受器细胞保护效应的研究[J].上海中医药杂志,2018,52(8):83-86.
[6] BIAN M,DU X,WANG P,et al.Combination of ginsenoside Rb1 and Rd protects the retina against bright light-induced degeneration[J].Sci Rep,2017,7(1):6015.
[7] JOLY S,PERNET V,DORFMAN A L,et al.Light-induced retinopathy:comparing adult and juvenile rats[J].Invest Ophthalmol Vis Sci,2006,47(7):3202-3212.
[8] ISHIKAWA F,OHGURO H,OHGURO I,et al. Prolonged rhodopsin phosphorylation in light-induced retinal degeneration in rat models[J].Invest Ophthalmol Vis Sci,2006,47(12):5204-5211.
[9]徐静,卞敏娟,崔金刚,等.白蒺藜对光损伤小鼠光感受器细胞的保护作用[J].眼科新进展,2017,37(2):110-113.
[10] BIAN M,DU X,CUI J,et al.Celastrol protects mouse retinas from bright light-induced degeneration through inhibition of oxidative stress and inflammation[J].J Neuroinflammation,2016,13(1):50.
[11] SZCZESNY P J,WALTHER P,MüLLER M.Light damage in rod outer segments:the effects of fixation on ultrastructural alteration[J]. Curr Eye Res,1996,15(8):807-814.
[12] VAN BEST J A,PUTTING B J,OOSTERHUIS J A,et al.Funetion and morphology of the retinal pigment epithelium after light-induced damage[J].Mierosc Res Tech,1997,36(2):77-88.
[13] ILIESCU D A,CIUBOTARU A,GHI爩M A,et al.Effect of sevoflurane preconditioning on light-induced retinal damage in diabetic rats[J].Rom J Ophthalmol,2018,62(1):24-33.