Amelioration of diabetic retinopathy in db/db mice by treatment with different proportional three active ingredients from Tibetan medicine Berberis dictyophylla F.
Amelioration of diabetic retinopathy in db/db mice by treatment with different proportional three active ingredients from Tibetan medicine Berberis dictyophylla F.
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DOI：
10.1016/j.jep.2021.114190
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发表时间：
2021-05
影响因子：
5.4
通讯作者：
Rui Li;Xiaopeng Ai;Ya Hou;Xianrong Lai;Xianli Meng;Xiaobo Wang
中科院分区：
文献类型：
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作者：
Rui Li;Xiaopeng Ai;Ya Hou;Xianrong Lai;Xianli Meng;Xiaobo Wang
民族药理研究著名藏药百里香，临床上用于防治糖尿病视网膜病变已有几千年的历史。本研究的目的是探讨黄连素、厚朴花碱和小檗胺对白叶小檗碱的协同保护作用及其作用机制。Db/db小鼠自发性视网膜损伤的研究材料与方法将db/db小鼠14周的自发性糖尿病视网膜病变模型随机分为8组：模型组、羟苯磺酸钙(CaDob，0.23 mg/kg)组和3种不同比例的白术有效成分1~6组。所有小鼠连续灌胃12周。记录并测量体重和空腹血糖(FBG)。苏木精-伊红染色和高碘酸-席夫(PAS)染色评价视网膜的病理改变和异常血管生成。采用双抗体夹心法测定血清中IL-6、H IF-1、α的水平。免疫荧光染色检测视网膜CD31、血管内皮生长因子、p-p38、p-JNK、p-ERK、NF-κB的蛋白表达。结果不同比例的三种有效成分对大鼠的体重无明显影响，但可降低空腹血糖，增加视网膜神经节细胞数，修复内界膜。PAS染色结果显示，药物治疗使视网膜血管基底膜变薄，内皮细胞/周细胞比例降低。此外，这些不同比例的有效成分均能显著下调视网膜CD31和血管内皮生长因子的蛋白水平，以及血清中缺氧诱导因子-1α和血管内皮生长因子的水平。视网膜血管内皮生长因子的基因表达也受到抑制。药物治疗可降低视网膜p-p38、p-JNK和p-ERK蛋白水平。药物治疗可逆转糖尿病小鼠视网膜前炎症因子NF-κB和血清IL-6的表达，逆转促凋亡基因bax的表达，增加抗凋亡基因bcl2的表达。结论不同比例的白术三种有效成分可改善db/db小鼠的DR。潜在的血管保护机制可能涉及抑制MAPK信号通路的磷酸化，从而减少炎症和凋亡事件。
Ethnopharmacological relevanceBerberis dictyophylla F., a famous Tibetan medicine, has been used to prevent and treat diabetic retinopathy (DR) for thousands of years in clinic. However, its underlying mechanisms remain unclear.Aim of the studyThe present study was designed to probe the synergistic protection and involved mechanisms of berberine, magnoflorine and berbamine fromBerberis dictyophyllaF. on the spontaneous retinal damage of db/db mice.Materials and methodsThe 14-week spontaneous model of DR in db/db mice were randomly divided into eight groups: model group, calcium dobesilate (CaDob, 0.23 g/kg) group and groups 1–6 (different proportional three active ingredients fromBerberis dictyophyllaF.). All mice were intragastrically administrated for a continuous 12 weeks. Body weight and fasting blood glucose (FBG) were recorded and measured. Hematoxylin-eosin and periodic acid–Schiff (PAS) stainings were employed to evaluate the pathological changes and abnormal angiogenesis of the retina. ELISA was performed to assess the levels of IL-6, HIF-1α and VEGF in the serum. Immunofluorescent staining was applied to detect the protein levels of CD31, VEGF, p-p38, p-JNK, p-ERK and NF-κB in retina. In addition, mRNA expression levels of VEGF, Bax and Bcl-2 in the retina were monitored by qRT-PCR analysis.ResultsTreatment with different proportional three active ingredients exerted no significant effect on the weight, but decreased the FBG, increased the number of retinal ganglionic cells and restored internal limiting membrane. The results of PAS staining demonstrated that the drug treatment decreased the ratio of endothelial cells to pericytes while thinned the basal membrane of retinal vessels. Moreover, these different proportional active ingredients can markedly downregulate the protein levels of retinal CD31 and VEGF, and serum HIF-1α and VEGF. The gene expression of retinal VEGF was also suppressed. The levels of retinal p-p38, p-JNK and p-ERK proteins were decreased by drug treatment. Finally, drug treatment reversed the proinflammatory factors of retinal NF-κB and serum IL-6, and proapoptotic Bax gene expression, while increased antiapoptotic Bcl-2 gene expression.ConclusionsThese results indicated that DR in db/db mice can be ameliorated by treatment with different proportional three active ingredients fromBerberis dictyophyllaF. The potential vascular protection mechanisms may be involved in inhibiting the phosphorylation of the MAPK signaling pathway, thus decreasing inflammatory and apoptotic events.