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吴凯,陈立浩,时健,刘倩宏,姚小磊.青光安颗粒剂对DBA/2J小鼠视神经中自噬途径相关蛋白表达的影响[J].湖南中医药大学学报,2022,42(4):536-541[点击复制] |
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| 青光安颗粒剂对DBA/2J小鼠视神经中自噬途径相关蛋白表达的影响 |
| 吴凯,陈立浩,时健,刘倩宏,姚小磊 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南中医药大学, 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;河北省眼科医院, 河北 邢台 054000;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) |
| 摘要: |
| 目的 研究青光安颗粒剂（以下简称为青光安）对自发性青光眼模型DBA/2J小鼠视神经中自噬途径相关蛋白表达的影响。方法 将10只C57BL/6J小鼠作为空白组；采用随机数字法将30只DBA/2J小鼠分为模型组、低剂量组、高剂量组，每组10只。喂养小鼠至38周龄，期间观测小鼠眼压，待DBA/2J小鼠自动成模后开始灌胃。模型组每日以生理盐水灌胃；低剂量组、高剂量组分别灌胃青光安20.75、103.75 g/kg，每日1次。各组小鼠均以常规饲料喂养。灌胃15 d后取右眼视神经。各组采用 Western blot法检测微管相关蛋白轻链3(microtubule-associated protein light chain 3, LC3)、溶酶体相关膜蛋白1(lysosome-associated membrane protein 1, LAMP1)、含半胱氨酸的天冬氨酸蛋白水解酶-3（cysteinyl aspartate specific proteinase, Caspase-3）、细胞色素C（cytochrome C, CytC）蛋白表达，采用RT-PCR法检测E3泛素连接酶（Parkin）、LAMP1、Caspase-3 mRNA的表达。结果 小鼠喂养至38周龄后，DBA/2J小鼠眼压上升，造模成功。与空白组相比，模型组LAMP1、Caspase-3、CytC、LC3-Ⅱ/LC3-Ⅰ蛋白表达升高，Parkin、LAMP1、Caspase-3 mRNA表达升高（P<0.05）。与模型组相比，低剂量组LAMP1、CytC、LC3-Ⅱ/LC3-Ⅰ蛋白表达降低（P<0.05），LAMP1、Caspase-3 mRNA表达降低（P<0.05）；高剂量组LAMP1、Caspase-3、CytC、LC3-Ⅱ/LC3-Ⅰ蛋白表达降低，Parkin、LAMP1、Caspase-3 mRNA表达降低（P<0.05）。与低剂量组相比，高剂量组Parkin、Caspase-3、LAMP1 mRNA表达水平降低（P<0.05）。结论 青光安可通过调控青光眼小鼠视神经中Parkin、LAMP1、LC3、Caspase-3、CytC的表达，使得线粒体自噬过程完整进行，对视神经具有保护作用。 |
| 关键词: 青光安颗粒剂 青光眼 线粒体自噬 视神经 DBA/2J小鼠 |
| DOI：10.3969/j.issn.1674－070X.2022.04.004 |
| 投稿时间：2021-05-01 |
| 基金项目:国家自然科学基金地区基金项目（81860870）；中国博士后科学基金面上资助项目（2018M640754）；全国中医药创新骨干人才培训项目（湘中医药函〔2019〕67号）；湖南省高层次卫生人才“225”工程培养项目（湘卫函〔2019〕196号）；中医药防治眼耳鼻咽喉疾病湖南省重点实验室开放基金项目（2018YZD02）。 |
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| Effect of Qingguang'an Granule on the expression of mitophagy pathway related proteins in optic nerve of DBA/2J mice |
| WU Kai,CHEN Lihao,SHI Jian,LIU Qianhong,YAO Xiaolei |
| (The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Provincial Key Laboratory of TCM Prevention and Treatment of Eye, Ear, Nose and Throat Diseases, Changsha, Hunan 410208, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hebei Eye Hospital, Xingtai, Hebei 054000, China;The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan Provincial Key Laboratory of TCM Prevention and Treatment of Eye, Ear, Nose and Throat Diseases, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To study the effect of Qingguang'an Granule on the expression of mitophagy pathway related proteins in the optic nerve of DBA/2J mice with spontaneous glaucoma. Methods Ten C57BL/6J mice were used as blank group. Thirty DBA/2J mice were randomly divided into model group, low-dose group and high-dose group, with 10 mice in each group by random number method. Mice were fed to 38 weeks of age, and intraocular pressure was observed during this period. After automatic modeling of DBA/2J mice, the mice were given intragastric administration. The model group was given normal saline intragastric administration every day. Qingguang'an Granule were gavaged at 20.75 g/kg and 103.75 g/kg in the low-dose group and high-dose group, once a day, respectively. Mice in each group were fed with conventional diet. The optic nerve of the right eye was removed 15 days after gavage. The protein expression levels of microtubuleassociated protein light chain 3 (LC3), lysosome-associated membrane protein 1 (LAMP1), cysteinyl aspartate specific proteinase (Caspase-3) and cytochrome C (CytC) in each group were detected by Western blot, and the expression levels of E3 ubiquitin ligase (Parkin), LAMP1 and Caspase-3 mRNA were detected by RT-PCR. Results After the mice were fed to 38 weeks of age, the intraocular pressure of DBA/2J mice increased, and the modeling was successful. Compared with blank group, protein expression levels of LAMP1, Caspase-3, CytC, LC3-Ⅱ/LC3-Ⅰ and expression levels of Parkin, LAMP1 and Caspase-3 mRNA were increased in model group (P<0.05). Compared with model group, the protein expression levels of LAMP1, CytC and LC3-Ⅱ/LC3-Ⅰ were decreased in low-dose group (P<0.05), and the expression levels of LAMP1 and Caspase-3 mRNA were decreased in low-dose group (P<0.05). The protein expression levels of LAMP1, Caspase-3, CytC, LC3-Ⅱ/LC3-Ⅰ and Parkin, LAMP1 and Caspase-3 mRNA were decreased in the high-dose group (P<0.05). Compared with the low-dose group, the expression levels of Parkin, Caspase-3 and LAMP1 mRNA in the high-dose group were decreased (P<0.05). Conclusion By regulating the expression of Parkin, LAMP1, LC3, Caspase-3 and CytC in the optic nerve of glaucoma mice, Qingguang'an Granule can make the mitochondrial autophagy process complete and has protective effect on the optic nerve. |
| Key words: Qingguang'an Granule glaucoma mitophagy optic nerve DBA/2J mice |
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