温阳益气活血方对遗传性视网膜色素变性小鼠感光细胞凋亡的影响及机制研究

目的观察中药温阳益气活血方对遗传性视网膜色素变性小鼠视网膜感光细胞凋亡的影响，并探寻其分子机制。方法将RDS（retinal degeneration slow）小鼠随机分为模型组和中药组，并设C57BL/6J小鼠为正常对照组，每组4雌2雄。中药组自雌雄合笼之日起开始给予温阳益气活血方（10 mg/g）灌胃，仔鼠出生后母鼠给予中药水煎液代替日常饮水，仔鼠出生7天开始给予中药水煎剂小剂量灌胃，出生21天按成年鼠剂量灌胃。模型组和正常组同时给予生理盐水灌胃。3组均于仔鼠出生后18、28、48天时采用视网膜电图（electroretinogram，ERG）检测视网膜功能，进行HE染色并计算外核层感光细胞层数，TUNEL法检测感光细胞凋亡率，免疫组织化学染色法计算视紫红质（Rhodopsin）及碱性成纤维细胞生长因子（basic fibroblast growth factor，bFGF）表达。结果与模型组比较，仔鼠出生后18天时，中药组最大混合反应ERG（Max-ERG） a、b波振幅及bFGF表达明显升高（P<0.05， P<0.01）；28、48天时，中药组Max-ERG a、b波振幅明显升高，外核层感光细胞层数明显增加，视网膜感光细胞凋亡率明显降低，Rhodopsin及bFGF表达明显升高，差异均有统计学意义（P<0.05， P<0.01）。结论温阳益气活血方可以有效抑制RDS小鼠感光细胞的凋亡，其机制可能与上调bFGF的表达有关。

英文摘要:

Objective To observe the effect of Wenyang Yiqi Huoxue Recipe （WYHR） on the apoptosis of photoreceptor cells in retinal degeneration slow （RDS） mice， and to investigate its molecular mechanisms. Methods RDS mice were randomly divided into the model group and the Chinese medicine group，and C57BL/6J mice were selected as the normal control group. Each group consisted of 4 female mice and 2 male mice. Mice in the Chinese medicine group were administered with WYHR （10 mg/g） by gastrogavage since mating. Baby mice drunk WYHR decoction instead of drinking water once they were born. The offspring were administrated with low dose WYHR decoction by gastrogavage from the 7th postnatal day， and the dose was increased to that for adult mice from the 21st postnatal day. Physiological saline was administrated to mice in the model group and the normal control group by gastrogavage. At 18， 28 and 48 postnatal days， electroretinogram （ERG） was used to evaluate the retina functional variation， and the apoptotic rate of photoreceptor cells was determined by TUNEL staining. HE staining was performed. The number of photoreceptor cells of the outer nuclear layer was calculated. Furthermore， effect of WYHR on Rhodopsin and basic fibroblast growth factor （bFGF） expression was examined using immunochemical assay. Results Compared with the model group， a-and b-wave latency and amplitude， as well as the bFGF expression sharply increased in the Max-ERG of the Chinese medicine group （P<0.05， P<0.01） at 18 postnatal days. At 28 and 48 postnatal days， a-and b-wave latency and amplitude sharply increased， photoreceptor cell layer numbers of the outer nuclear layer obviously increased， the apoptosis rate of retinal photoreceptor cells obviously decreased， expressions of Rhodopsin and bFGF in the Chinese medicine group significantly increased （P<0.05， P<0.01）. Conclusion WYHR could effectively inhibit the apoptosis of photoreceptor cells in RDS mice， which might be attributed to up-regulating bFGF expression.

关闭