蒋鹏飞 欧晨 彭俊 姚震 田野 杨毅敬 宋厚盼 徐剑 彭清华

〔摘要〕 目的 觀察枸杞子-丹参对rd10小鼠视网膜Caspase-12前体、Caspase-2表达的影响。方法 将40只rd10小鼠随机分为5组，空白组灌胃生理盐水;对照组灌胃维生素A溶剂;枸杞组灌胃枸杞子中药超微配方颗粒溶液;丹参组灌胃丹参中药超微配方颗粒溶液;枸杞加丹参组（杞参组）灌胃枸杞子加丹参中药超微配方颗粒溶液;8只C57小鼠为野生组，灌胃生理盐水。给药28 d后，RT-qPCR检测小鼠视网膜Caspase-12前体、Caspase-2 mRNA表达情况，免疫组化、免疫荧光法检测小鼠视网膜Caspase-12前体、Caspase-2蛋白表达情况。结果 RT-qPCR检测显示：除杞参组外，各组小鼠视网膜Caspase-12前体、Caspase-2 mRNA相对表达量高于野生组，差异有统计学意义（P<0.05）;杞参组小鼠视网膜Caspase-12前体、Caspase-2 mRNA相对表达量低于空白组、对照组，差异均有统计学意义（P<0.05）。免疫组化检测显示：各组小鼠视网膜Caspase-12前体、Caspase-2平均光密度高于野生组，差异均有统计学意义（P<0.05）;杞参组Caspase-12前体、Caspase-2蛋白平均光密度低于空白组、对照组、枸杞组、丹参组，差异均有统计学意义（P<0.05）。免疫荧光检测显示：各组小鼠视网膜Caspase-12前体、Caspase-2蛋白平均荧光强度高于野生组，差异均有统计学意义（P<0.05）;杞参组Caspase-12前体、Caspase-2蛋白平均荧光强度低于空白组、对照组、枸杞组、丹参组，差异均有统计学意义（P<0.05）。结论 枸杞子-丹参可以抑制rd10小鼠视网膜Caspase-12前体、Caspase-2 mRNA和蛋白的表达，从而抑制感光细胞的凋亡，维持视网膜正常功能结构，保护视功能。

〔关键词〕 视网膜色素变性;细胞凋亡;枸杞子;丹参Caspase-2;Caspase-12前体

〔中图分类号〕R285.5       〔文献标志码〕A       〔文章编号〕doi：10.3969/j.issn.1674-070X.2021.04.003

〔Abstract〕 Objective To observe the effect of Gouqizi （Lycii Fructus） and Danshen （Salviae Miltiorrhizae Radix Et Rhizoma） on the expression of Caspase-12 precursor and Caspase-2 in rd10 mice. Methods 40 rd10 mice were randomly divided into 5 groups. The blank group was given distilled water; the control group was given intragastric vitamin A solvent; the Gouqizi （Lycii Fructus） group was given intragastric Gouqizi （Lycii Fructus） Chinese medicine ultrafine granular solution; the Danshen （Salviae Miltiorrhizae Radix Et Rhizoma） group was given intragastric Danshen （Salviae Miltiorrhizae Radix Et Rhizoma） Chinese medicine ultrafine formula granule solution; Gouqizi （Lycii Fructus） plus Danshen （Salviae Miltiorrhizae Radix Et Rhizoma） group （QiShen group） was given intragastric Gouqizi （Lycii Fructus） plus Danshen （Salviae Miltiorrhizae Radix Et Rhizoma） Chinese medicine superfine granule solution; 8 C57 mice as the wild group， was given intragastric distilled water. 28 days after the administration， RT-qPCR was used to detect the expression of Caspase-12 precursor and Caspase-2 mRNA in the retina of rd10 mice， and the expression of Caspase-12 precursor and Caspase-2 proteins in the retina was detected by immunohistochemistry and immunofluor？鄄escence. Results The RT-qPCR test showed that except for the QiShen group， the relative expression levels of Caspase-12 precursor and Caspase-2 mRNA in the retina of each group were higher than that of the wild group， and the differences were statistically significant （P<0.05）; the relative expression levels of Caspase-12 precursor and Caspase-2 mRNA in the retina of the QiShen group were lower than that of the blank group and the control group， the differences were statistically significant （P<0.05）. Immunohistochemical detection showed that the average optical density of Caspase-12 precursor and Caspase-2 in the retina of mice in each group was higher than that of the wild group， and the differences were statistically significant （P<0.05）; the average optical density of Caspase-12 precursor and Caspase-2 of the QiShen group was lower than that of the blank group， control group， Gouqizi （Lycii Fructus） group， and Danshen （Salviae Miltiorrhizae Radix Et Rhizoma） group， and the differences were statistically significant （P<0.05）. Immunofluorescence detection showed that the average fluorescence intensity of Caspase-12 precursor and Caspase-2 in the retina of mice in each group was higher than that in the wild group， and the differences were statistically significant （P<0.05）; Caspase-12 precursor and Caspase-2 average fluorescence intensity of QiShen group was lower than that of the blank group， control group， Gouqizi （Lycii Fructus） group and Danshen （Salviae Miltiorrhizae Radix Et Rhizoma） group， and the differences were statistically significant （P<0.05）. Conclusion Gouqizi （Lycii Fructus） and Danshen （Salviae Miltiorrhizae Radix Et Rhizoma） can inhibit the expression of rd10 mice Caspase-12 precursors， Caspase-2， thereby inhibiting the apoptosis of photoreceptor cells， maintaining normal functional structure of retina， protecting view function.

〔Keywords〕 retinitis pigmentosa; apoptosis; Gouqizi （Lycii Fructus）; Danshen （Salviae Miltiorrhizae Radix Et Rhizoma）; Caspase-2; Caspase-12 precursor

视网膜色素变性（retinitis pigmentosa， RP）是临床常见的视网膜疾病，以视网膜色素变化为特征，病至后期，可对视力、视野造成严重影响[1]。我国人群发病率约为1/4 000，男女比例约3∶2[2]。多数学者认为RP视力损失的主要原因是视网膜感光细胞的凋亡[3-5]，凋亡是细胞程序化死亡方式的一种，本研究以视网膜色素变性小鼠rd10小鼠为研究对象，通过观察枸杞子-丹参对rd10小鼠视网膜感光细胞凋亡相关因子Caspase-12前体和Caspase-2的影响，探讨枸杞子丹参治疗RP的机制。

1 材料

1.1  实验动物

40只4周龄SPF级rd10小鼠，雌雄各半，质量18～20 g，购自美国Jackson（JAX）实验室，证书编号：191A11353;8只4周龄SPF级C57小鼠，雌雄各半，质量18～20 g，由湖南斯莱克景达公司提供，证书编号：43004700062254。实验动物均饲养于湖南斯莱克景达公司动物实验中心SPF房，室温（22±2） ℃，湿度50%～55%，定期更换垫料，自由饮食。……