碘酸钠诱发视网膜色素变性大鼠模型的生理指标和组织病理学分析

doi:10.12300/j.issn.1674-5817.2022.149

实验动物与比较医学 ›› 2023, Vol. 43 ›› Issue (2): 124-135.DOI: 10.12300/j.issn.1674-5817.2022.149

碘酸钠诱发视网膜色素变性大鼠模型的生理指标和组织病理学分析

谭莹¹^,²(), 廖文平¹^,², 高启龙³, 李勇¹^,², 史新辉¹^,², 王京昆¹^,²()()

^1.云南省药物研究所, 昆明 650011
^2.云南省中药和民族药新药创制企业重点实验室, 昆明 650011
^3.昆明市中医医院, 昆明 650031

收稿日期:2022-09-22 修回日期:2023-02-17 出版日期:2023-04-25 发布日期:2023-04-25
通讯作者: 王京昆（1969—），男，学士，正高级工程师，研究方向：新药研发和药物安全性评价研究。E-mail： wjkyimm@163.com。ORCID： 0009-0006-9724-6286
作者简介:谭莹（1979—），女，博士，正高级工程师，研究方向：药物安全性评价研究。E-mail： ty1203@163.com。ORCID： 0009-0007-3084-9020
基金资助:
云南省重大科技专项“灯盏花口服类制剂新适应症和毒性作用系统研究”(2017ZF002-08)

Physiological Indexes and Histopathology Analysis of Sodium Iodate-Induced Retinitis Pigmentosa in Rats

Ying TAN¹^,²(), Wenping LIAO¹^,², Qilong GAO³, Yong LI¹^,², Xinhui SHI¹^,², Jingkun WANG¹^,²()()

^1.Yunnan Institute of Materia Medica, Kunming 650011, China
^2.Yunnan Province Company Key Laboratory for TCM and Ethnic Drug of New Drug, Kunming 650011, China
^3.Kunming Traditional Chinese Medicine Hospital, Kunming 650031, China

Received:2022-09-22 Revised:2023-02-17 Published:2023-04-25 Online:2023-04-25
Correspondence to: WANG Jingkun (ORCID: 0009-0006-9724-6286), E-mail: wjkyimm@163.com

摘要/Abstract

摘要：

目的全面评价碘酸钠腹腔注射对SD大鼠生理指标和视网膜组织病理学特征的影响。方法选取64只SD大鼠随机分为阴性对照组和模型组，每组32只，雌雄各半。模型组以10 mL/kg剂量腹腔注射碘酸钠（6 mg/mL）1次，阴性对照组以10 mL/kg剂量腹腔注射生理盐水注射液1次。每天观察大鼠一般体征，注射当天（0天）和注射后第2、6、9、13、16、20、23、27、29、36、43、50、57天称量所有存活大鼠体质量；注射后第3、7、21、28、41、62天随机选取大鼠（第28天为12只/组，其余为4只/组，均为雌雄各半）进行血清生化学指标检测，系统解剖摘取脏器称重，然后对主要脏器和组织进行HE染色，以及眼HE和TUNEL染色；注射后第28、62天进行血常规指标检测。结果注射碘酸钠后，模型组88%的大鼠出现一过性稀便，观察期间大鼠体质量增长稍缓且以雄鼠较明显。注射后第28天模型组雌鼠红细胞容积（red blood cell volume，RDW）和雄鼠尿素氮（urea nitrogen，BUN）、网织红细胞计数（reticulocyte count，Retic#）及百分比（reticulocyte percentage，Retic%）均升高，与阴性对照组比较的差异均有统计学意义（均P＜0.05）。雌雄鼠白细胞（white blood cells，WBC）、红细胞（red blood cells，RBC）、血红蛋白（hemoglobin，HGB）、红细胞比容（hematocrit，HCT）均有降低趋势，但组间差异均没有统计学意义（均P＞0.05）。模型组雄鼠胸腺重量、系数除注射后第7天大于阴性对照组外，其余时间点均小于阴性对照组，但组间差异均没有统计学意义（均P＞0.05）。组织病理学检查可见模型组大鼠视网膜由波浪状改变逐步发展为异常心电图样改变，各层排列紊乱，外核层局灶性变薄，视网膜外核层细胞凋亡；病变发生率、病变评分和凋亡细胞数量均明显高于或多于同时期的阴性对照组，第28天的组间差异有统计学意义（均P＜0.01）。结论碘酸钠腹腔注射除可使大鼠视网膜变性外，对血清生化、血常规部分指标也有一定程度的影响，还出现大鼠体质量增长略缓慢和一过性粪便性状改变的现象。因此，在使用该模型开展药物安全性评价时，应关注造模试剂对动物的影响。

关键词: 碘酸钠, 生理指标, 视网膜组织病理, 视网膜色素变性模型, 大鼠

Abstract:

Objective To evaluate the effects of intraperitoneal injection of sodium iodate on the physiological indexes and retinal histopathological characteristics of SD rats comprehensively. Methods A total of 64 rats were randomly divided into negative control group and model group, half male and half female. The model group was intraperitoneally injected with 6 mg/mL sodium iodate once at the dose of 10 mL/kg and the negative control group was injected with 10 mL/kg 0.9% NaCl once. The body weight of all surviving rats was detected on the day of injection (0 day) and the 2nd, 6th, 9th, 13th, 16th, 20th, 23rd, 27th, 29th, 36th, 43rd, 50th and 57th days after injection. On the 3rd, 7th, 21st, 28th, 41st and 62nd days after injection, the rats were randomly selected (12 rats in each group on the 28th day, and 4 rats in each group at other time points, those in each group were half male and half female) for serum biochemical indexes detection. The organs were dissected and weighed, and then the main organs and tissues were stained with HE, and the eyes were stained with HE and TUNEL. Blood routine indexes were detected on the 28th and 62nd day after injection. Results After injection of sodium iodate, 88% of the rats in the model group had transient loose stools. During the observation period, the body weight of the rats increased slightly and was more obvious in male rats. On the 28th day after injection, compared with the negative control group, the red blood cell volume (RDW) of female rats and blood urea nitrogen (BUN), reticulocyte count (Retic#) and reticulocyte percentage (Retic%) of male rats in the model group increased significantly (all P＜0.05). The white blood cell (WBC), red blood cell (RBC), hemoglobin (HGB) and hematocrit (HCT) of male and female rats showed decreasing trends, but there were no significant differences between the two groups (all P >0.05). The thymus weight and coefficient of male rats in the model group were smaller than those in the negative control group except for the 7th day after injection, but there were no significant differences between the two groups (all P ＞0.05). Histopathological examination showed that the retina of the model group gradually developed from wavy changes to abnormal electrocardiogram (ECG)-like changes, with disordered arrangement of each layer, focal thinning of the outer nuclear layer, and apoptosis of the outer nuclear layer of the retina. The incidence of lesions, lesion score and the number of apoptotic cells in the model group were significantly higher than or more than those in the negative control group at the same time, and the difference between the groups on the 28th day was statistically significant (all P＜0.01). Conclusion In addition to retinal degeneration in rats, intraperitoneal injection of sodium iodate also had a certain degree of influence on serum biochemical and blood routine indexes, and also showed a slight slow growth of body weight and transient changes in fecal traits. Therefore, when using this model to evaluate drug safety, the effects of modeling reagents on animals should be paid to attention.

Key words: Sodium iodate, Physiological indexes, Retinal histopathology, Retinis pigmentosa model, Rats

中图分类号:

Q95-33

谭莹,廖文平,高启龙,等. 碘酸钠诱发视网膜色素变性大鼠模型的生理指标和组织病理学分析[J]. 实验动物与比较医学, 2023, 43(2): 124-135. DOI: 10.12300/j.issn.1674-5817.2022.149.

Ying TAN,Wenping LIAO,Qilong GAO,et al. Physiological Indexes and Histopathology Analysis of Sodium Iodate-Induced Retinitis Pigmentosa in Rats[J]. Laboratory Animal and Comparative Medicine, 2023, 43(2): 124-135. DOI: 10.12300/j.issn.1674-5817.2022.149.

图/表 6

注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别16、16、14、12、12、12、12、10、10、4、4、2、2、2只；与阴性对照组间比较，^?P＜0.05。
">

图1 碘酸钠注射后雌雄鼠体质量变化曲线图
注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别16、16、14、12、12、12、12、10、10、4、4、2、2、2只；与阴性对照组间比较，^?P＜0.05。

Figure 1 Body weight curve of male and female rats after sodium iodate injection
Note：The model group was intraperitoneally injected with 6 mg/mL sodium iodate solution once at the dose of 10 mL/kg；the negative control group was intraperitoneally injected with normal saline at the same volume and frequency. The number of rats in the two groups at each time point was 16， 16， 14， 12，12， 12， 12， 10， 10， 4， 4， 2， 2， 2 respectively for single sex. Compared with the negative control group， ^?P＜0.05.

注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只；与阴性对照组比较，^?P＜0.05。
">

图2 碘酸钠注射后雌雄鼠血清生化指标变化
注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只；与阴性对照组比较，^?P＜0.05。

Figure 2 Changes of serum biochemical indexes of male and female rats after sodium iodate injection
Note： The model group was intraperitoneally injected with 6 mg/mL sodium iodate solution once at the dose of 10 mL/kg； the negative control group was intraperitoneally injected with normal saline at the same volume and frequency. The number of rats in the two groups at each time point was 2， 2， 2， 6， 2， 2 respectively for single sex. Compared with the negative control group， ^?P＜0.05.

注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只；与阴性对照组间比较，^??P＜0.01。
">

图3 碘酸钠注射后雌雄鼠血常规指标变化
注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只；与阴性对照组间比较，^??P＜0.01。

Figure 3 Changes of blood routine indexes of male and female rats after sodium iodate injection
Note：The model group was intraperitoneally injected with 6 mg/mL sodium iodate solution once at the dose of 10 mL/kg；the negative control group was intraperitoneally injected with normal saline at the same volume and frequency. The number of rats in the two groups at each time point was 2， 2， 2， 6， 2， 2 respectively for single sex. Compared with the negative control group，^??P＜0.01.

注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只。
">

图4 碘酸钠注射后雌雄鼠各脏器质量及系数变化图
注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只。

Figure 4 Changes of organ weight and coefficient of male and female rats after sodium iodate injection
Note：The model group was intraperitoneally injected with 6 mg/mL sodium iodate solution once at the dose of 10 mL/kg；the negative control group was intraperitoneally injected with normal saline at the same volume and frequency. The number of rats in the two groups at each time point was 2， 2， 2， 6， 2， 2 respectively for single sex.

注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只。a示模型组注射后第3天视网膜呈波浪样改变；b示模型组注射后第7天视网膜呈异常心电图样改变，各层排列紊乱；图c～f分别示模型组注射后第21、28、41、62天视网膜呈异常心电图样改变，各层排列紊乱、外核层局灶变薄；g示阴性对照组视网膜正常；h-i为两组大鼠的视网膜病变发生率及HE评分比较（病变1为视网膜呈波浪样改变，病变2为视网膜呈心电图样改变）与阴性对照组比较，^??P<0.01。图中比例尺大小均为50 μm。
">

图5 碘酸钠注射后不同时间点的大鼠眼视网膜病变情况（HE染色， ×400）
注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只。a示模型组注射后第3天视网膜呈波浪样改变；b示模型组注射后第7天视网膜呈异常心电图样改变，各层排列紊乱；图c～f分别示模型组注射后第21、28、41、62天视网膜呈异常心电图样改变，各层排列紊乱、外核层局灶变薄；g示阴性对照组视网膜正常；h-i为两组大鼠的视网膜病变发生率及HE评分比较（病变1为视网膜呈波浪样改变，病变2为视网膜呈心电图样改变）与阴性对照组比较，^??P<0.01。图中比例尺大小均为50 μm。

Figure 5 Retinopathy in rats at different time points after sodium iodate injection （HE staining， ×400）
Note：The model group was intraperitoneally injected with 6 mg/mL sodium iodate solution once at the dose of 10 mL/kg； the negative control group was intraperitoneally injected with normal saline at the same volume and frequency. The number of rats in the two groups at each time point was 2， 2， 2， 6， 2， 2 respectively for single sex. a shows the retina of the model group appeared wave-like changes on the 3rd day after injection； b shows the retina of the model group appeared abnormal electrocardiogram （ECG）-like changes on the 7th day after injection， and the arrangement of each layer was disordered； c-f show the retina of the model group appeared abnormal ECG-like changes on the 21st， 28th， 41st and 62nd days after injection， with disordered arrangement of each layer and focal thinning of the outer nuclear layer. g shows normal retina in negative control group； h-i show the comparison of the incidence of retinopathy and HE score between the two groups （Lesion 1 shows wave-like changes in the retina， lesion 2 shows ECG-like changes in the retina）. ^??P<0.01， vs the negative control group. The scale size in the figure are 50 μm.

注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只。a～f分别示模型组注射后第3、7、21、28、41、62天视网膜外核层细胞TUNEL染色阳性（箭头所示为TUNEL染色阳性细胞）；g示阴性对照组视网膜TUNEL染色阴性；h为两组大鼠的视网膜外核层TUNEL染色阳性细胞数比较，与阴性对照组比较，^??P<0.01。图中比例尺大小均为50 μm。
">

图6 碘酸钠注射后不同时间点的大鼠眼视网膜TUNEL染色情况（×400）
注：模型组一次性腹腔注射质量浓度为6 mg/mL碘酸钠溶液10 mL/kg；阴性对照组以相同容积和频次腹腔注射生理盐水注射液。各时间点两组大鼠数量分别为单性别2、2、2、6、2、2只。a～f分别示模型组注射后第3、7、21、28、41、62天视网膜外核层细胞TUNEL染色阳性（箭头所示为TUNEL染色阳性细胞）；g示阴性对照组视网膜TUNEL染色阴性；h为两组大鼠的视网膜外核层TUNEL染色阳性细胞数比较，与阴性对照组比较，^??P<0.01。图中比例尺大小均为50 μm。

Figure 6 TUNEL staining of rat retina at different time points after sodium iodate injection （×400 ）
Note：The model group was intraperitoneally injected with 6 mg/mL sodium iodate solution once at the dose of 10 mL/kg；the negative control group was intraperitoneally injected with normal saline at the same volume and frequency. The number of rats in the two groups at each time point was 2， 2， 2， 6， 2， 2 respectively for single sex. a-f show that TUNEL staining was positive in the outer nuclear layer of the retina on the 3rd， 7th， 21st， 28th， 41st and 62nd days after injection in the model group （the arrows show TUNEL staining positive cells）； g shows that the TUNEL staining of the retina in the negative control group was negative； h shows the comparison of the number of TUNEL staining positive cells in the outer nuclear layer of the two groups of rats. ^??P<0.01， vs the negative control group. The scale size in the figure are 50 μm.

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碘酸钠诱发视网膜色素变性大鼠模型的生理指标和组织病理学分析

Physiological Indexes and Histopathology Analysis of Sodium Iodate-Induced Retinitis Pigmentosa in Rats

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