Garriz Angela, Morokuma Junji, Bowman Maytal, Pagni Sarah, Zoukhri Driss
Department of Comprehensive Care, Tufts University School of Dental Medicine, Boston, MA, USA.
Public Health and Community Service, Tufts University School of Dental Medicine, Boston, MA, USA.
Front Ophthalmol (Lausanne). 2022;2. doi: 10.3389/fopht.2022.873486. Epub 2022 Apr 27.
In the lacrimal gland, myoepithelial cells (MEC) express muscle contractile proteins such as alpha smooth muscle actin (SMA) and calponin and therefore can contract to help expel lacrimal fluid. In a previous study, we demonstrated that lacrimal gland MEC express the oxytocin receptor (OXTR) and they contract under oxytocin (OXT) stimulation. Using NOD and MRL/lpr mice (animal models of Sjogren's syndrome), we reported a decrease in SMA and calponin protein levels plus a decline in acini contraction after stimulation with OXT. It is known that proinflammatory cytokines, such as interleukin-1β (IL-1β), tumor necrosis factor alpha (TNF-α) or interferon gamma (IFN-γ), can affect OXTR expression and signaling capacity and inhibit MEC contraction. The aim of the current study was to investigate if proinflammatory cytokines are implicated in the loss of MEC contractile ability. Thus, lacrimal gland MEC from a SMA-GFP transgenic mouse were treated with IL-1β (10 ng/ml) for a total of 7 days. At days 0, 2, 4 and 7, GFP intensity, cell size/area, contractile proteins amounts and MEC contraction were assessed. At day 0, control and treated cells showed no differences in GFP intensity and cell size. GFP intensity started to decrease in treated MEC at day 2 (20%; p=0.02), continuing after day 4 (25%; p=0.007) and 7 (30%; p=0.0001). Mean cell area was also reduced at day 2 (34%; p=0.0005), and after 4 (51%; p<0.0001) and 7 days (30%; p=0.0015). The contraction assay at day 2 showed a 70% decrease of contraction in treated MEC (p<0.0001), 73% (p<0.0001) at day 4 and 82% (p=0.0015) at day 7 when compared to control. Levels of contractile proteins were measured on day 7 showing a decrease in SMA and calponin amount in treated MEC compared with the control group (around 30%; p=0.0016 and p=0.0206; respectively). Similar results were observed when TNF-α and IFN-γ were added along with IL-1β. Taken together the present data and those from our previous studies with Sjogren's syndrome mouse models, they strongly suggest that proinflammatory cytokines affect lacrimal gland MEC contractile ability that may account for the reduced tear secretion associated with Sjogren's syndrome dry eye disease.
在泪腺中,肌上皮细胞(MEC)表达肌肉收缩蛋白,如α平滑肌肌动蛋白(SMA)和钙调蛋白,因此能够收缩以帮助排出泪液。在先前的一项研究中,我们证明泪腺MEC表达催产素受体(OXTR),并且它们在催产素(OXT)刺激下会收缩。使用NOD和MRL/lpr小鼠(干燥综合征的动物模型),我们报道了用OXT刺激后SMA和钙调蛋白水平降低以及腺泡收缩能力下降。已知促炎细胞因子,如白细胞介素-1β(IL-1β)、肿瘤坏死因子α(TNF-α)或干扰素γ(IFN-γ),可影响OXTR表达和信号传导能力并抑制MEC收缩。本研究的目的是调查促炎细胞因子是否与MEC收缩能力丧失有关。因此,将来自SMA-GFP转基因小鼠的泪腺MEC用IL-1β(10 ng/ml)处理总共7天。在第0、2、4和7天,评估GFP强度、细胞大小/面积、收缩蛋白量和MEC收缩情况。在第0天,对照细胞和处理细胞在GFP强度和细胞大小方面没有差异。处理后的MEC在第2天GFP强度开始下降(20%;p=0.02),在第4天(25%;p=0.007)和第7天(30%;p=0.0001)继续下降。平均细胞面积在第2天也减少(34%;p=0.0005),在第4天(51%;p<0.0001)和第7天(30%;p=0.0015)后减少。与对照相比,第2天的收缩试验显示处理后的MEC收缩减少70%(p<0.0001),第4天减少73%(p<0.0001),第7天减少82%(p=0.0015)。在第7天测量收缩蛋白水平,显示处理后的MEC中SMA和钙调蛋白量与对照组相比减少(分别约为30%;p=0.001和p=0.0206)。当TNF-α和IFN-γ与IL-1β一起添加时观察到类似结果。综合目前的数据以及我们先前对干燥综合征小鼠模型的研究数据,强烈表明促炎细胞因子影响泪腺MEC收缩能力,这可能是干燥综合征干眼症相关泪液分泌减少的原因。