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陈立浩,时健,刘倩宏,姚小磊,蒋鹏飞.菊花总黄酮对去势所致干眼雄兔泪腺组织中AR、AR mRNA、Bax mRNA及形态学的影响[J].湖南中医药大学学报,2020,40(4):406-411[点击复制] |
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| 菊花总黄酮对去势所致干眼雄兔泪腺组织中AR、AR mRNA、Bax mRNA及形态学的影响 |
| 陈立浩,时健,刘倩宏,姚小磊,蒋鹏飞 |
| (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;湖南中医药大学第一附属医院, 湖南 长沙 410007) |
| 摘要: |
| 目的 考察菊花总黄酮治疗去势雄兔干眼的具体机制。方法 采取随机数字法将150只雄兔分为15组，编号分别为A1、A3、A5、B1、B3、B5、C1、C3、C5、D1、D3、D5、E1、E3、E5，每组10只，A-E组中序号对应干预时间，其中A组不做任何干预处理，B组行假去势术，C、D、E组行双侧去势术建立雄激素水平缺乏型干眼模型。A、B、C组均用生理盐水灌胃，D组行雄激素（丙酸睾酮）肌肉注射，E组用菊花总黄酮灌胃。分别于治疗后的1、3、5月，将动物处死。观察：（1）雄激素样效应以免疫组织化学法测定泪腺组织中雄激素受体（androgen receptor，AR）光密度，辅以RT-PCR技术测定AR mRNA表达，以观察泪腺组织中AR上调情况；（2）细胞凋亡以RT-PCR技术测定泪腺中细胞凋亡相关基因蛋白Bax mRNA的表达；（3）形态学改变使用透射电镜观察泪腺超微结构变化。结果 （1）雄激素样效应：E组与C组相比，其泪腺组织中AR明显上调（P<0.01），AR mRNA表达率明显增长（P<0.01）；（2）细胞凋亡：E组Bax mRNA表达率与C组比较下降明显（P<0.01）；（3）形态学改变：E组在干预后，线粒体肿胀减轻，丢失或成空泡变，而C组线粒体肿胀，局灶凋亡坏死，凋亡明显。结论 菊花总黄酮具有较好的雄激素样效应，能够抑制Bax mRNA凋亡，恢复泪腺组织形态。它将有望应用于由雄激素水平下降所致干眼的临床治疗中。 |
| 关键词: 干眼 去势雄兔 菊花总黄酮 雄激素样效应 Bax mRNA 形态学 |
| DOI：10.3969/j.issn.1674-070X.2020.04.005 |
| 投稿时间：2019-12-30 |
| 基金项目:国家自然科学基金地区基金项目（81260550）；国家中医药管理局中医眼科学重点学科建设项目（ZK1801YK015）；中医药防治五官科疾病湖南省重点实验室建设项目（2017TP1018）；湖南省中医五官科学重点学科建设项目；湖南中医药大学中医学国内一流建设学科项目。 |
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| Effects of Total Flavonoids from Chrysanthemum on AR, AR mRNA, Bax mRNA and Morphology in Lacrimal Gland of Castrated Dry Eye Male Rabbits |
| CHEN Lihao,SHI Jian,LIU Qianhong,YAO Xiaolei,JIANG Pengfei |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Key Laboratory of Traditional Chinese Medicine for Prevention and Treatment of Eye, Ear, Nose and Throat Diseases in Hunan Province, Changsha, Hunan 410208, China;The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
| Abstract: |
| Objective To explore the specific mechanism of total flavonoids from chrysanthemum in the treatment of castrated male rabbit dry eyes. Methods A total of 150 male rabbits were randomly divided into 15 groups with random number method, and numbered A1, A3, A5, B1, B3, B5, C1, C3, C5, D1, D3, D5, E1, E3, E5, with 10 rabbits in each group. There was no intervention in group A. Sham castration was given in group B. Bilateral castration was performed in groups C, D and E. Group A, B and C were given normal saline. Group D was given testosterone propionate intramuscular injection, and group E was given total flavonoids of chrysanthemum by gavage. The animals were killed 1, 3 and 5 months after treatment. (1) For the observation of androgen like effect,androgenreceptor(AR) density in lacrimal gland was measured by immunohistochemistry, and AR mRNA expression was measured by RT-PCR to observe the up-regulation of AR in lacrimal gland. (2) For the observation of apoptosis, Bax mRNA expression in lacrimal gland was measured by RT-PCR. (3) For the observation of morphological changes, ultrastructural structural changes of lacrimal gland were observed by transmission electron microscopy. Results (1) androgen like effect:compared with the model group C, the AR in lacrimal gland of group E was significantly up-regulated (P<0.01), and the AR mRNA expression rate was significantly increased (P<0.01). (2) Apoptosis:the Bax mRNA expression rate of group E was significantly decreased (P<0.01) compared with that of group C. (3) Morphological changes:after the intervention, the swelling of mitochondria in Group E was reduced, and the mitochondria in group C were lost or vacuolized, whilethe swelling of mitochondria, focal apoptosis and necrosis in group C was obvious. Conclusion The total flavone from chrysanthemum has a better androgen like effect, which can inhibit Bax mRNA apoptosis and restore lacrimal gland morphology. It is expected to be used in the clinical treatment of dry eye caused by the decrease of androgen level. |
| Key words: dry eye disease castrated male rabbit total flavones from chrysanthemum androgen-like effect Bax mRNA morphology |
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