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目的:考察夏枯草水提物(PV)对链脲霉素(STZ)诱导的C57小鼠非增殖性糖尿病视网膜病(NPDR)的改善作用及其作用机制。方法:通过连续5 d腹腔注射STZ制备C57小鼠糖尿病模型,进而诱导建立糖尿病并发症NPDR实验动物模型。将造模成功的小鼠按体质量随机分为模型组、PV低剂量组(100 mg/kg)、PV高剂量组(200 mg/kg),另设有一组小鼠腹腔注射溶媒对照作为正常对照组,每组16只。小鼠末次腹腔注射STZ后1个月灌胃给药PV或生理盐水,连续1个月。采用视网膜组织伊文氏蓝渗漏实验检测NPDR小鼠中血-视网膜屏障(BRB)的破坏情况;采用ELISA方法检测血清中白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)的含量;采用Western-blot方法检测视网膜组织中小胶质细胞/巨噬细胞特异性蛋白(Iba1)的表达,并检测核因子κB(NF-κB)的转录激活。结果:模型组小鼠视网膜中伊文氏蓝渗漏值明显升高(P<0.001),而PV高低剂量组均对此有显著的改善作用(P<0.001);模型组小鼠血清中IL-1β、TNF-α含量明显高于正常组(P<0.05),而PV高低剂量组血清中IL-1β、TNF-α含量均较模型组明显降低(P<0.05,P<0.01,P<0.001)。模型组小鼠视网膜组织中Iba-1蛋白表达明显上调(P<0.001),而PV高低剂量组Iba-1表达明显低于模型组(P<0.001)。模型组小鼠视网膜组织中Phospho-p65蛋白含量显著增加(P<0.001),胞核中p65蛋白的表达显著上调(P<0.001);而PV高低剂量组中磷酸化p65蛋白表达明显低于模型组(P<0.05,P<0.001),胞核中p65蛋白表达明显低于模型组(P<0.001)。结论:PV能改善STZ诱导C57小鼠NPDR,其机制可能是抑制了NF-κB介导的炎性信号通路,减少了炎性因子IL-1β、TNF-α的表达,缓解了STZ诱导糖尿病小鼠视网膜组织的炎性损伤和BRB的渗漏。
Objective: To investigate the effect of Prunella vulgaris aqueous extract (PV) on streptozotocin (STZ) -induced non-proliferative diabetic retinopathy (NPDR) induced by C57 in mice and its mechanism. Methods: The C57 mouse model of diabetes was established by intraperitoneal injection of STZ for 5 consecutive days, which led to the establishment of experimental animal model of diabetic NPDR. The successful mice were randomly divided into model group, PV low dose group (100 mg / kg) and PV high dose group (200 mg / kg). Another group of mice were given intraperitoneal injection of vehicle control as normal Control group, 16 rats in each group. One month after the last intraperitoneal injection of STZ, mice were given PV or normal saline orally for 1 month. The damage of blood-retinal barrier (BRB) in NPDR mice was detected by Evans blue leak test of retinal tissue. The levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF- The expression of microglobulin / macrophage-specific protein (Iba1) in retinal tissue was detected by Western-blot and the transcriptional activation of nuclear factor-kappa B (NF-κB) was detected. Results: The infiltration of Evans Blue in the retina of the model group was significantly increased (P <0.001), while both the high dose and low dose PV groups had a significant improvement (P <0.001). IL- The levels of IL-1β and TNF-α in serum of PV high and low dose groups were significantly lower than those of model group (P <0.05, P <0.01, P <0.001) ). The expression of Iba-1 protein in retina of model group was significantly increased (P <0.001), while the expression of Iba-1 in PV high and low dose group was significantly lower than that of model group (P <0.001). The phosphorylation of p65 protein in retina was significantly increased (P <0.001) in model group, while the expression of p65 protein in nucleus was significantly increased (P <0.001). The phosphorylation of p65 protein in PV group was lower than that in model Group (P <0.05, P <0.001). The expression of p65 protein in nucleus was significantly lower than that in model group (P <0.001). CONCLUSION: PV can improve the STZ-induced NPDR in C57 mice, which may be related to the inhibition of NF-κB-mediated inflammatory signaling pathway and the decrease of inflammatory cytokines IL-1β and TNF-α Inflammatory damage of rat retinal tissue and leakage of BRB.