枸杞多糖通过抑制NF-κB/NLRP3通路减少视网膜色素变性小鼠感光细胞凋亡

目的探讨枸杞多糖可否通过抑制核因子激活的B细胞的κ-轻链增强子/NOD样受体热蛋白结构域相关蛋白3(NF-κB/NLRP3)信号通路减少视网膜色素变性(RP)小鼠视网膜光感受器细胞的凋亡。方法(1)体外实验将小鼠视网膜神经节细胞(661W细胞)分为正常组、模型组、枸杞多糖低剂量组(40 mg/L)、中剂量组(80 mg/L)、高剂量组(160 mg/L)和阳性药物对照组(NLRP3抑制剂,160 mg/L);分别用50、100、200和400μmol/L四种剂量的H_(2)O_(2)干预661W细胞,选出最佳H_(2)O_(2)浓度(200μmol/L)造模,细胞计数试剂盒CCK-8测定细胞活力,流式细胞仪检测细胞凋亡率,免疫荧光检测NLRP3标志物的表达,酶联免疫吸附试验(ELISA)和免疫印迹法(WB)检测细胞凋亡标志物的表达。(2)选用C57/BL6小鼠和Rd10小鼠进行体内实验,分组为正常组、模型组、枸杞多糖低剂量组(0.08 g/d)、中剂量组(0.16 g/d)、高剂量组(0.32 g/d)和阳性药物对照组(NLRP3抑制剂,0.08 g/d),其中正常组为C57/BL6小鼠,其余组为Rd10小鼠,每组10只,相对应的药物连续灌胃4周,通过视网膜电图、组织病理学检查和WB等方法观察NF-κB/NLRP3通路及细胞凋亡标志物的表达,评估枸杞多糖对视网膜光感受器细胞凋亡的影响。结果(1)体外实验中,与正常组相比,模型组661W细胞凋亡率明显增加(P<0.01),且NF-κB/NLRP通路关键蛋白NLRP3、NF-κB、p-NF-κB和促凋亡蛋白caspase-3的表达水平上调(P<0.01),Bax/Bcl-2的比值增大(P<0.01),白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α的浓度显著升高(P<0.01)。与模型组相比,高剂量的枸杞多糖降低了661W细胞的凋亡率(P<0.01),下调了NF-κB/NLRP3通路关键蛋白NF-κB、NLRP3、p-NF-κB和caspase-3的表达(P<0.01),减小了Bax/Bcl-2的比值(P<0.01),降低了IL-1β和TNF-α的浓度(P<0.01)。(2)体内实验中,高剂量的枸杞多糖显著增加了Rd10小鼠外核层(ONL)厚度的形态变化以及a波和b波振幅的功能变化(P<0.01),下调了NF-κB(P<0.05)、NLRP3、p-NF-κB和caspase-3的表达水平(P<0.01),减小了Bax/Bcl-2的比值(P<0.01),降低了IL-1β(P<0.01)和TNF-α(P<0.05)的浓度。结论枸杞多糖可能通过抑制NF-κB/NLRP3通路改善视网膜形态和功能,保护光感受器免受细胞凋亡的影响。

复明片对实验性视网膜脱离时及复位后突触素表达的影响

目的研究复明片对视网膜脱离时及复位后视网膜突触素（synalatophysin，SY）表达的影响。方法将48只家兔随机分为4组，分别为正常组、模型组、西药对照组及复明片组，采用视网膜下腔注射透明质酸钠的方法建立视网膜脱离模型，10～14d后视网膜自动复位。正常组、模型组给予生理盐水，西药对照组给予西药混合溶液，复明片组给予复明片混悬液灌胃，分别于术后1周（视网膜脱离时）及3周（视网膜复位后）采用免疫组化法观察SY表达情况。结果视网膜脱离导致SY表达显著减少，术后1周时，模型组、西药对照组及复明片组SY染色0D值分别为0、07±0，叭、0、08±0．01、0.13±0．02，较正常组（0．24±0．02）明显降低（P〈0．01）；3周时，SY表达较视网膜脱离时增加，3组染色OD值分别为0、13±0．01、0．14±0，02、0．20±0、02，但与正常组（0，25±0，03）比较，其染色OD值仍显著降低（P〈0．01；P〈0，01；P〈0．05）；在视网膜脱离时及复位后2个时相，复明片组SY染色OD值均高于相同时间点的模型组和西药对照组，差异亦有极显著性（P〈0.01）。结论视网膜脱离导致视功能严重受损，而复明片可促进视网膜神经元突触密度和分布的恢复（轴突再生），促进视觉信息的传递，从而促进视功能的恢复。

复明片对实验性视网膜脱离复位细胞凋亡的影响

目的:研究复明片对视网膜脱离时及复位后细胞凋亡的影响。方法:将48只家兔随机分为4组,每组12只,采用视网膜下腔注射透明质酸钠的方法建立视网膜脱离动物模型,10～14天后视网膜自动复位。正常组、模型组予生理盐水,西药对照组予西药混合溶液,复明片组予复明片混悬液灌胃,分别于术后1、3周采用TUNEL法观察视网膜的细胞凋亡情况。结果:术后1周和术后3周,复明片组视网膜细胞凋亡较相同时间点模型组及西药对照组明显减少,细胞凋亡指数显著降低(P<0.01)。结论:复明片能减少网脱所致视网膜细胞凋亡,从而提高视功能。

密蒙花提取物治疗兔去势所致干眼症

目的观察密蒙花提取物对去势所致的干眼症雄兔基础泪液分泌量,泪腺局部炎症因子转化生长因子β1(transforming growth factor-β1,TGF-β1)、白细胞介素1β(interleukin-1β,IL-1β)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)的影响,探讨密蒙花提取物抗雄激素水平下降所致的干眼症的作用机制。方法将30只日本大耳雄兔随机分为空白组(A)、模型组(B)、1倍剂量密蒙花提取物治疗组(C)、2倍剂量密蒙花提取物治疗组(D)和染料木黄酮治疗组(E),每组6只,对B、C、D、E组行雄激素去势术建立动物模型,对C、D、E组分别以密蒙花提取物1倍剂量和2倍剂量以及染料木黄酮连续灌胃1个月,对全部实验兔行SchirmerI试验,采用免疫组织化学方法,对兔泪腺组织中炎症因子TGF-β1、IL-1β、TNF-α进行检测。结果C、D、E组SchirmerI试验测量值明显高于B组(P<0.01),在泪腺导管及腺泡上皮细胞中,IL-1β、TNF-α阳性表达的细胞数均明显低于B组,而TGF-β1阳性表达的细胞数均明显高于B组(P<0.01)。以上检测结果C、D组均优于E组(P<0.05)。结论密蒙花提取物中主要成分为黄酮类物质,可显著抑制雄激素水平降低后兔干眼症的发生。其作用机制可能与黄酮类物质化学结构与雄激素类似,可起到拟雄激素效应,从而调节泪腺局部炎症反应有关。密蒙花提取物可能成为治疗干眼症的新途径。

Dan Huang Ming Mu Recipe Suppresses the Progression of Streptozotocin-induced Diabetic Retinopathy After Retinal Laser Photocoagulation in Brown Norway Rats via Down-regulating Vascular Endothelial Growth Factor and Up-regulating Pigment Epithelium-derived

Objective To analyze the effects of Dan Huang Ming Mu Recipe(DHMMR)(a pharmaceutical preparation from herbs and having the function of replenishing vital essence,removing heat,promoting blood circulation and excreting pathogenic water) on diabetic retinopathy(DR) after retinal laser photocoagulation through regulating the expression of vascular endothelial growth factor(VEGF) and pigment epithelium-derived factor(PEDF).Methods Forty male Brown Norway(BN) rats were randomly divided into blank group(group A,10 BN rats) and model group(30 BN rats).DR models were induced by 40 mg/kg streptozotocin(STZ) and the body weight and blood glucose of rats were monitored.After 12-week injection,fluorescein fundus angiography(FFA) and histopathological examination were detected to confirm the successful establishment of DR models.Subsequently,the right eyes of model group rats were conducted retinal laser photocoagulation with the left eyes having no retinal laser photocoagulation and rats of the model group were randomly divided into group B(the model control group),group C(the positive control group),and group D(the DHMMR group),with 10 rats in each group.Rats of the group A and the group B were given vehicle,and the group C were given calcium dobesilate suspension by gavage,and the group D were given DHMMR by gavage.After 4-week gavage,FFA was carried out to observe the fundus microvascular change.Then,the rats were sacrificed to do histopathological examinations and the serum levels of P-selectin,cAMP,cGMP,and the relative expression of the protein of VEGF and PEDF in retina were detected.Results After 12-week STZ injection,the blood glucose of the model group were pronouncedly higher than the blank group(P < 0.01).Fundus micro-hemangioma changes were observed in the rats of the model group through FFA,and the rats of the blank group did not see any changes.The pictures of HE staining showed that the retinal structure of the model group was more disordered than the blank group.After 4-week gavage,treatments with DHMMR showed dramatic reduction of serum levels of Pselectin,cAMP and cGMP compared with the group B(P < 0.01).FFA and histopathological examinations of DHMMR-treated rats revealed significantly suppression of retinal edema,fundus microvascular destruction and retinal destruction distinguishing from the group B.And the relative expression of VEGF protein of the group D and the group A was markedly lower than that of the group B(P < 0.01).The relative expression of PEDF protein of the group D and the group A was dramatically higher than that of the group B to the contrary(P < 0.01).Conclusions DHMMR demonstrates pronounced suppressive effects on the progression of DR after retinal laser photocoagulation,through down-regulating VEGF and upregulating PEDF.

有些视疲劳是脖子闹的

出现视物模糊等表象时,人们常以为是自己的眼睛出了问题,一般情况下,只要到眼科进行仔细检查,大多能够找到原因,通过适当治疗可治愈或缓解。然而,有部分视疲劳患者,虽然有明显的视疲劳症状,但是眼部查不出明显的原因,这就要怀疑是否是颈椎病导致了视疲劳。