滋阴明目丸对RCS大鼠视网膜Fas/FasL表达的影响
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摘要
目的观察滋阴明目丸对RCS大鼠视网膜Fas/FasL表达的影响。方法选择RCS大鼠,(1.51±0.27)月龄,共24只,按随机数字表法将其分为三组:空白组、模型组、滋阴明目丸组(雌雄各4只,n=8)。空白组:RCS(rdy~(+/+),p~(+/+))大鼠,灌胃生理盐水;模型组:RCS(rdy~(-/-),p~(-/-))大鼠,灌胃生理盐水;滋阴明目丸组:RCS(rdy~(-/-),p~(-/-))大鼠,灌胃滋阴明目丸。灌胃30 d后,免疫组化染色观察标本视网膜各层结构的形态学变化,Western blot法测定RCS大鼠视网膜Fas/FasL表达水平。结果免疫组化染色结果显示:空白组大鼠视网膜各层结构清晰;模型组大鼠视网膜的厚度低于空白组,且各层结构不清晰;滋阴明目丸组大鼠视网膜厚度低于空白组但高于模型组,视网膜各层结构较清晰。Western blot法检测结果显示:滋阴明目丸组大鼠视网膜Fas、FasL蛋白相对表达量明显低于模型组,差异有高度统计学意义(P <0.01)。结论滋阴明目丸对视网膜的超微结构具有保护作用,能抑制视网膜上Fas/FasL的表达,从而减轻视网膜感光细胞的凋亡,达到保护视细胞的目的。
Objective To observe the effect of Ziyin Mingmu Pills on the expression of Fas/FasL in the retina of RCS rats. Methods The RCS rats were selected, aged(1.51±0.27) months, total 24 rats, they were randomly divided into 3 groups: blank group, model group, Ziyin Mingmu Pills group(4 male and 4 female, n = 8). Blank group: RCS(rdy~(+/+), p~(+/+))rats were given normal saline by intragastric administration; model group: RCS(rdy~(-/-), p~(-/-)) rats were given normal saline by intragastric administration; Ziyin Mingmu Pills group: RCS(rdy~(-/-), p~(-/-)) rats were given Ziyin Mingmu Pills by intragastric administration. After intragastric administration for 30 d, the morphological changes of each layer of the retina sample were observed by immunohistochemistry. The expression levels of Fas/FasL in the retina of rats were determined by Western blot. Results The results of immunohistochemistry showed that each layer of retinal structure of the rats in the blank group was clear; the thickness of retina in the model group was lower than that in the blank group and the structure of each layer of retina was not clear; the retinal thickness of Ziyin Mingmu Pills group was lower than that of blank group but higher than that of blank group, the structure of each layer of retina was clear. The results of Western blot assay showed that the relative expression of Fas, FasL protein in the retina of Ziyin Mingmu Pills group was lower than that in the model group, the difference was highly statistically significant(P < 0.01). Conclusion Ziyin Mingmu Pills has a protective effect on the ultrastructure of retina, which can inhibit the expression of Fas/FasL of the retina and reduce the apoptosis of retinal photoreceptor cells, thus achieving the purpose of protecting visual cells.
Objective To observe the effect of Ziyin Mingmu Pills on the expression of Fas/FasL in the retina of RCS rats. Methods The RCS rats were selected, aged(1.51±0.27) months, total 24 rats, they were randomly divided into 3 groups: blank group, model group, Ziyin Mingmu Pills group(4 male and 4 female, n = 8). Blank group: RCS(rdy~(+/+), p~(+/+))rats were given normal saline by intragastric administration; model group: RCS(rdy~(-/-), p~(-/-)) rats were given normal saline by intragastric administration; Ziyin Mingmu Pills group: RCS(rdy~(-/-), p~(-/-)) rats were given Ziyin Mingmu Pills by intragastric administration. After intragastric administration for 30 d, the morphological changes of each layer of the retina sample were observed by immunohistochemistry. The expression levels of Fas/FasL in the retina of rats were determined by Western blot. Results The results of immunohistochemistry showed that each layer of retinal structure of the rats in the blank group was clear; the thickness of retina in the model group was lower than that in the blank group and the structure of each layer of retina was not clear; the retinal thickness of Ziyin Mingmu Pills group was lower than that of blank group but higher than that of blank group, the structure of each layer of retina was clear. The results of Western blot assay showed that the relative expression of Fas, FasL protein in the retina of Ziyin Mingmu Pills group was lower than that in the model group, the difference was highly statistically significant(P < 0.01). Conclusion Ziyin Mingmu Pills has a protective effect on the ultrastructure of retina, which can inhibit the expression of Fas/FasL of the retina and reduce the apoptosis of retinal photoreceptor cells, thus achieving the purpose of protecting visual cells.
引文
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[13]Sanchez-Bretano A,Janjua U,Gargini G,et al.Melatonin protects 661W cells from cell death induced by H2O2via inhibition of the Fas/FasL-Caspase 3 pathway[J].Invest Ophthalmol Vis Sci,2017,58(8):354.
[14]Xia HL,Li CJ,Hou XF,et al.Interferon-γaffects leukemia cell apoptosis through regulating Fas/FasL signaling pathway[J].Eur Rev Med Pharmacol Sci,2017,21(9):2244-2248.
[15]Tao H,Lu L,Xia Y,et al.Antitumor effector B cells directly kill tumor cells via the Fas/FasL pathway and are regulated by IL-10[J].Eur J Immunol,2015,45(4):999-1009.
[16]Hu X,He D,Zhou C,et al.Marginatoxin induces human hepatoma BEL-7402 cells apoptosis in vitro and in vivo via activation of Fas/FasL-mediated apoptotic pathway[J].Biomed Res,2017,28(3):1242-1246.
[17]Zhang JF,Shi LL,Zhang L,et al.MicroRNA-25 negatively regulates cerebral ischemia/reperfusion injury-induced cell apoptosis through Fas/FasL pathway[J].J Mol Neurosci,2016,58(4):507-516.
[18]Matsumoto H,Murakami Y,Kataoka K,et al.Membranebound and soluble Fas ligands have opposite functions in photoreceptor cell death following separation from the retinal pigment epithelium[J].Cell Death Dis,2015,6:e1986.
[19]Kurnia I,Siregar B,Soetopo S,et al.KORELASI ANTARAMIB-1,AgNOR DAN APOPTOSIS Caspase-3 DENGANRESPONS KEMORADIOTERAPI PADA KANKERSERVIK[J].JSTNI,2013,14(1):51-64.
[20]窦仁慧,金明.视网膜色素变性中医治疗及研究进展[J].中国中医眼科杂志,2011,21(1):59-61.
[21]邓婷婷,窦仁慧,潘琳,等.温阳益气活血方对遗传性视网膜色素变性小鼠感光细胞凋亡的影响及机制研究[J].中国中西医结合杂志,2013,33(8):1122-1128.
[22]徐赵钕,章仕淼,刘玲玲,等.明日地黄丸对糖尿病视网膜病变大鼠细胞自噬及Akt-mTOR通路的影响[J].中国医药导报,2018,15(20):16-20,32.
[23]孙河,樊晓瑞.针药并用治疗视网膜色素变性临床研究[J].针灸临床杂志,2010,26(6):20-22.
[24]帅天姣,代海燕,陈丹丹,等.β-榄香烯对STZ致糖尿病性视网膜病变大鼠增殖期SIRT1和VEGF表达的影响[J].中国现代医生,2018,56(21):38-40.
[25]胡楚璇,李穗华,张霞,等.人参皂苷Rg1对大鼠视神经损伤的保护作用研究[J].中国医药导报,2018,15(3):17-21.
[26]刘相和,迟焕芳.枸杞子提取液对RCS大鼠遗传性视网膜变性的作用[J].齐鲁医学杂志,2009,24(2):119-120.
[27]李雪丽,唐由之,范吉平,等.补肾益精方对RCS大鼠视网膜变性损伤的保护作用研究[J].中国中医眼科杂志,2016,26(3):144-149.
[2]Sacchetti M,Mantelli F,Rocco ML,et al.Recombinant human nerve growth factor treatment promotes photoreceptor survival in the retinas of rats with retinitis pigmentosa[J].Curr Eye Res,2017,42(7):1064-1068.
[3]Roddy GW,Yasumura D,Matthes MT,et al.Long-term photoreceptor rescue in two rodent models of retinitis pigmentosa by adeno-associated virus delivery of Stanniocalcin-1[J].Exp Eye Res,2017,165:175-181.
[4]张盼盼,董琪,华英彬,等.贝伐单抗对人视网膜色素上皮细胞形态,凋亡率及凋亡相关因子表达的影响[J].眼科新进展,2018,38(4):314-318.
[5]徐剑.基于RHO、XBP1、Caspase12表达探讨枸杞、丹参对虚中夹瘀证RP模型大鼠的干预研究[D].长沙:湖南中医药大学,2016.
[6]Ishikawa M,Sawada Y,Yoshitomi T.Structure and function of the interphotoreceptor matrix surrounding retinal photoreceptor cells[J].Exp Eye Res,2015,133:3-18.
[7]Meier P,Finch A,Evan G.Apoptosis in development[J].Nature,2000,407(6805):796-801.
[8]叶河江,王莹,张露,等.针刺对感光细胞凋亡大鼠视网膜形态学的影响[J].辽宁中医杂志,2012,39(9):1857-1859.
[9]Hong LK,Chen Y,Smith CC,et al.CD30-Redirected Chimeric Antigen Receptor T Cells Target CD30+and CD30-Embryonal Carcinoma via Antigen-Dependent and Fas/FasLInteractions[J].Cancer Immunol Res,2018,6(10):1274-1287.
[10]Du P,Li SJ,Ojcius DM,et al.A novel Fas-binding outer membrane protein and lipopolysaccharide of Leptospira interrogans induce macrophage apoptosis through the Fas/FasL-caspase-8/-3 pathway[J].Emerg Microbes Infec,2018,7(1):135.
[11]Potter CS,Silva KA,Kennedy VE,et al.Loss of FAS/FASLsignalling does not reduce apoptosis in Sharpin null mice[J].Exp Dermatol,2017,26(9):820-822.
[12]Svandova EB,Vesela B,Lesot H,et al.Expression of Fas,FasL,caspase-8 and other factors of the extrinsic apoptotic pathway during the onset of interdigital tissue elimination[J].Histochem Cell Biol,2017,147(4):497-510.
[13]Sanchez-Bretano A,Janjua U,Gargini G,et al.Melatonin protects 661W cells from cell death induced by H2O2via inhibition of the Fas/FasL-Caspase 3 pathway[J].Invest Ophthalmol Vis Sci,2017,58(8):354.
[14]Xia HL,Li CJ,Hou XF,et al.Interferon-γaffects leukemia cell apoptosis through regulating Fas/FasL signaling pathway[J].Eur Rev Med Pharmacol Sci,2017,21(9):2244-2248.
[15]Tao H,Lu L,Xia Y,et al.Antitumor effector B cells directly kill tumor cells via the Fas/FasL pathway and are regulated by IL-10[J].Eur J Immunol,2015,45(4):999-1009.
[16]Hu X,He D,Zhou C,et al.Marginatoxin induces human hepatoma BEL-7402 cells apoptosis in vitro and in vivo via activation of Fas/FasL-mediated apoptotic pathway[J].Biomed Res,2017,28(3):1242-1246.
[17]Zhang JF,Shi LL,Zhang L,et al.MicroRNA-25 negatively regulates cerebral ischemia/reperfusion injury-induced cell apoptosis through Fas/FasL pathway[J].J Mol Neurosci,2016,58(4):507-516.
[18]Matsumoto H,Murakami Y,Kataoka K,et al.Membranebound and soluble Fas ligands have opposite functions in photoreceptor cell death following separation from the retinal pigment epithelium[J].Cell Death Dis,2015,6:e1986.
[19]Kurnia I,Siregar B,Soetopo S,et al.KORELASI ANTARAMIB-1,AgNOR DAN APOPTOSIS Caspase-3 DENGANRESPONS KEMORADIOTERAPI PADA KANKERSERVIK[J].JSTNI,2013,14(1):51-64.
[20]窦仁慧,金明.视网膜色素变性中医治疗及研究进展[J].中国中医眼科杂志,2011,21(1):59-61.
[21]邓婷婷,窦仁慧,潘琳,等.温阳益气活血方对遗传性视网膜色素变性小鼠感光细胞凋亡的影响及机制研究[J].中国中西医结合杂志,2013,33(8):1122-1128.
[22]徐赵钕,章仕淼,刘玲玲,等.明日地黄丸对糖尿病视网膜病变大鼠细胞自噬及Akt-mTOR通路的影响[J].中国医药导报,2018,15(20):16-20,32.
[23]孙河,樊晓瑞.针药并用治疗视网膜色素变性临床研究[J].针灸临床杂志,2010,26(6):20-22.
[24]帅天姣,代海燕,陈丹丹,等.β-榄香烯对STZ致糖尿病性视网膜病变大鼠增殖期SIRT1和VEGF表达的影响[J].中国现代医生,2018,56(21):38-40.
[25]胡楚璇,李穗华,张霞,等.人参皂苷Rg1对大鼠视神经损伤的保护作用研究[J].中国医药导报,2018,15(3):17-21.
[26]刘相和,迟焕芳.枸杞子提取液对RCS大鼠遗传性视网膜变性的作用[J].齐鲁医学杂志,2009,24(2):119-120.
[27]李雪丽,唐由之,范吉平,等.补肾益精方对RCS大鼠视网膜变性损伤的保护作用研究[J].中国中医眼科杂志,2016,26(3):144-149.