石斛夜光丸光感受器细胞保护效应的研究

Protective effects of Shihu Yeguang Pill on photoreceptor cells

目的探讨石斛夜光丸抗光损伤诱导光感受器细胞退行性改变的效应。方法采用BALB/c小鼠,分为正常对照组、模型组以及石斛夜光丸不同时间点给药组。模型组及石斛夜光丸治疗组接受白炽光光照30 min[光照强度为(10 000±10)lx]以建立视网膜光损伤模型。石斛夜光丸治疗组连续给药7 d,光照前暗适应24 h,治疗组分别在光照前立即给药或光照前30 min给予石斛夜光丸水煎液200μl灌胃,正常对照组和模型组给予等量0.9%NaCl溶液灌胃。光照后7 d通过光学相干断层扫描(OCT)检测小鼠视网膜形态结构。采用石蜡切片进行苏木素-伊红(HE)染色和视紫红质(RHO)、中波敏感视蛋白(M-opsin)免疫组织化学染色,以进一步明确视网膜形态学及视杆细胞、视锥细胞的病理改变情况。结果正常对照组视网膜结构完整,RHO和M-opsin表达分布正常;与正常对照组相比,模型组外核层显著变薄,光感受器细胞外节层及内节层结构紊乱,RHO和M-opsin表达显著降低;与模型组相比,石斛夜光丸两给药组视网膜形态基本完整清晰,RHO和M-opsin的表达增多。结论石斛夜光丸具有显著的抗光损伤诱导的光感受器细胞退行性改变的作用。

Objective To investigate the protective effects of Shihu Yeguang Pill against light-induced photoreceptor degenerative changes. Methods BALB/c mice were randomly divided into the normal control group,model group and Shihu Yeguang Pill treatment groups at different time points. The retinal light damage model was established in the model group and Shihu Yeguang Pill treatment groups by incandescent light at intensity of（10 000±10） lx lux for 30 min. The Shihu Yeguang Pill treatment groups were treated for continuous 7 days,and 24 hours before illumination the mice were received dark adaptation. The treatment groups were treated with decoction of Shihu Yeguang Pill by intragastric administration at volume of 200 μl immediately or 30 minutes before illumination. The normal control group and model group were treated with 0. 9% Na Cl solution at equivalent volume. 7 days after illumination,the optical coherence tomography（OCT）imaging was performed to detect the retinal structure of mice. The paraffin sections were made to perform the hematoxylin and eosin（HE） staining and immunohistochemical assessment of rhodopsin（RHO） and mid-wavelength sensitive cone opsin（M-opsin）,in order to further clarify the changes of retinal morphology and patho logical changes of rod cells and cone cells. Results The retinal structures in the normal control group were intact,and the expressions and disposition of RHO and M-opsin were normal. Compared with the normal control group,the outer nuclear layer was obviously thinner in the model group,the structures of outer and inner segment layers of photoreceptor were disorganized and the expressions of RHO and M-opsin were remarkably decreased. Compared with the model group,the retinal structures were nearly intact and clear in Shihu Yeguang Pill treatment groups,and the expressions of RHO and M-opsin were increased. Conclusion Shihu Yeguang Pill shows significant protective effects against light-induced photoreceptor degenerative changes.