Dias Lara Cristina, Zheng Changyu, Murashima Adriana de Andrade Batista, Dias Ana Carolina, Fantucci Marina Zilio, Nominato Luiz Fernando, da Silva Lilian Eslaine Costa Mendes, Rocha Eduardo Melani
Ribeirão Preto Medical School, University of São Paulo, Ribeirao Preto, SP, Brazil.
Molecular Physiology and Therapeutic Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health Bethesda, Bethesda, Maryland, USA.
Curr Eye Res. 2021 Sep;46(9):1314-1319. doi: 10.1080/02713683.2021.1893754. Epub 2021 Mar 30.
: The aims of this work were a) to describe the histology of the lacrimal gland (LG) and cornea induced by an adenovirus (Ad) vector encoding the human erythropoietin (Epo) gene delivered to the LG and b) to evaluate the therapeutic potential of this strategy to prevent benzalkonium chloride (BAK) corneal toxicity.: Structure and function of male Wistar rats LG were compared in the groups: 1) naïve control and 2) Ad-hEpo in the right LG (RLG). The protective response against BAK eye drops was compared among the groups 1) naïve control, 2) BAK in the right eye, 3) Ad-hEpo RLG + BAK and 4) Ad-hEpo in the right salivary gland (RSG)+BAK. Ad-hEpo groups received an injection of AdLTR2EF1a-hEPO (25 ul, 10 particles/ml) in the right LG or SG (positive control). The BAK groups received 0.2% BAK in the right cornea twice a day. The tests applied after 7 days, included tear secretion, hEPO mRNA detection by qRT-PCR, LG and cornea histology, LG ELISA for cytokines and hematocrit.: hEPO mRNA was present in the Ad-hEpo RLG and RSG, but not kidney or liver samples (negative controls). TNF-α and IL-1β increased in the LG exposed to Ad-hEpo compared to naïve control ( = .0115 and = .0397, respectively). BAK reduced tear secretion, but this reduction was prevented by Ad-hEpo RLG+BAK and Ad-hEpo RSG+BAK ( = .017). The corneal epithelia were thinner in the BAK-treated groups independent of Ad-hEpo ( = .0009). Hematocrit increased only in the Ad-hEpo RSG group ( = .01).: Ad-hEpo infection of rat LG and SG induces local, but only the SG infection induced systemic changes in rats. Importantly, Ad-hEpo attenuated the BAK-mediated toxic reduction in tear flow. Future studies must consider viral vector tissue tropism, biodistribution and effective therapeutic gene products for ocular surface diseases.
a）描述将编码人促红细胞生成素（Epo）基因的腺病毒（Ad）载体注射到泪腺（LG）后，泪腺和角膜的组织学变化；b）评估该策略预防苯扎氯铵（BAK）所致角膜毒性的治疗潜力。比较以下几组雄性Wistar大鼠LG的结构和功能：1）未处理对照组；2）右侧LG注射Ad-hEpo组。比较以下几组对BAK滴眼液的保护性反应：1）未处理对照组；2）右眼滴注BAK组；3）右侧LG注射Ad-hEpo + BAK组；4）右侧唾液腺（RSG）注射Ad-hEpo + BAK组。Ad-hEpo组在右侧LG或SG（阳性对照）注射AdLTR2EF1a-hEPO（25 μl，10颗粒/ml）。BAK组在右侧角膜每天滴注两次0.2% BAK。7天后进行的检测包括泪液分泌、通过qRT-PCR检测hEPO mRNA、LG和角膜组织学检查、LG细胞因子ELISA检测和血细胞比容检测。hEPO mRNA存在于右侧LG注射Ad-hEpo组和右侧SG注射Ad-hEpo组，但在肾脏或肝脏样本（阴性对照）中未检测到。与未处理对照组相比，暴露于Ad-hEpo的LG中TNF-α和IL-1β增加（分别为P = 0.0115和P = 0.0397）。BAK减少泪液分泌，但右侧LG注射Ad-hEpo + BAK组和右侧SG注射Ad-hEpo + BAK组可预防这种减少（P = 0.017）。在BAK处理组中，无论是否注射Ad-hEpo，角膜上皮均变薄（P = 0.0009）。仅右侧SG注射Ad-hEpo组的血细胞比容增加（P = 0.01）。大鼠LG和SG感染Ad-hEpo可引起局部变化，但只有SG感染可引起大鼠全身变化。重要的是，Ad-hEpo减轻了BAK介导的泪液分泌减少。未来的研究必须考虑病毒载体的组织嗜性、生物分布以及用于眼表疾病的有效治疗性基因产物。
