Microvesicles (MVs) is small membrane vesicles released from different cell types under different conditions. Studies have shown that MVs may mediate vascular inflammation, angiogenesis, and other pathological processes. MVs may play an important role in the pathogenesis of diabetic retinopathy (DR) by mediating endothelial cell injury, thrombosis and neovascularization. The plasma MV level may be an effective parameter to monitor the development of DR. This article will summarize the research progress of the relationship between MVs and DR in recent years.
The classical Hippo pathway leads to the phosphorylation of downstream effector molecules Hippo-Yes-associated protein (Yap) and transcriptional coactivator PDZ-binding motif (Taz) serine sites through a kinase response, thereby promoting cell proliferation, controlling cell polarity, changing cytoskeleton, it plays an important regulatory role in various pathophysiological processes such as epithelial-mesenchymal transition and inhibition of cell contact. Studies have shown that Yap/Taz can affect the progression of vitreoretinal diseases, opening up new prospects for the pathogenesis and clinical treatment of diabetic retinopathy, proliferative vitreoretinopathy, and retinal ischemia-reperfusion injury. Exploring the molecular mechanism of Yap/Taz provides a possible therapeutic target for future research in the treatment of retinal fibrosis diseases such as diabetic retinopathy and proliferative vitreoretinopathy. At the same time, regulating the activity of local Yap/Taz in the retina will also become an effective therapeutic target for damage-repair in retinal ischemia-reperfusion injury. However, Yap inhibitors have potential retinal toxicity and are still in the preclinical development stage. Further research on the mechanism of action and clinical safety of Yap inhibitors will provide new methods for the treatment of retinal diseases.
Rituximab (RTX) is a monoclonal antibody directed against the CD20 antigen expressed on B cells. It has been successfully employed in the treatment of non-Hodgkin's lymphoma and varied systemic autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, and granulomatosis with polyangiitis. Recently its efficacy in the treatment of ocular inflammatory diseases (OID), including refractory scleritis, peripheral ulcerative keratitis, uveitis, and ocular cicatricial pemphigoid, has aroused more concerns. The literature suggests that RTX may be useful for controlling the inflammation and decreasing or stopping the use of corticosteroids and other immunosuppressants in OID, which may contribute a new treatment alternative in patients with the recalcitrant and sight-threatening forms of OID. This article reviews the clinical application status of RTX in the treatment of OID.
ObjectiveTo study how CD73 is shed from the retinal pigment epithelium (RPE) surface.MethodsCD73 shedding was induced by treating RPE with lipopolysaccharides (LPS) and TNF-α. After Phospholipase C (PLC) or pan matrix metalloproteinase (MMP) inhibitors were added, surface amount of CD73 was evaluated by flow cytometry (FACS). Then selective inhibitors or their corresponding siRNAs of MMP-2 and MMP-9 were applied to the treatments of RPE; and their effects on induced CD73 shedding were evaluated by FACS. By site directed mutagenesis, mutations were introduced to Lys547-Phe548 coding sites of CD73 cDNA, which was cloned in a pcDNA mammalian expression vector. Both wt-CD73 and mutated-CD73 were over expressed in CD73-/- RPE and their induced shedding was compared.ResultsLPS and TNF-α induced CD73 shedding from RPE was completely blocked by the addition of pan MMP inhibitor but not PLC inhibitor. Selective MPP-9, but not MMP-2, inhibitor or its siRNA blocked CD73 shedding. In CD73-/- RPE induced CD73 shedding was happened to overexpressed wt-CD73 but not Lys547-Phe548 sites mutant CD73.ConclusionMMP-9 is responsible for shedding CD73 from RPE through hydrolyzing its Lys547 -Phe548 sites.
ObjectiveTo investigate the changes of choroid thickness in adolescents with different types of non-pathological high myopia (HM). MethodsA retrospective observational study. From January 2021 to April 2022, 179 eyes of 101 adolescents with myopia in Liaocheng Aier Eye Hospital were collected and analyzed. According to the spherical equivalent (SE) and corneal curvature, subjects were divided into mild myopia or emmetical eye group (control group), HM group, occult HM group (OHM group) and super HM group (SHM group). There were 52 eyes in 30 cases, 47 eyes in 26 cases, 42 eyes in 24 cases and 38 eyes in 21 cases, respectively. Medical optometry, intraocular pressure, optical coherence tomography (OCT), axial length (AL) and corneal curvature were measured. The macular foveal choroidal thickness was analyzed by using spectral-domain OCT. The diopter was expressed in SE. The thickness of choroid in the fovea of macular region was measured by enhanced deep imaging with frequency domain OCT. The thickness of choroid was measured in 9 regions within 1 mm, 3 mm from the fovea, including the upper, lower, nasal and temporal regions. Generalized estimating equation was used to compare the data among groups, and the least significant difference t-test was used to compare the data among groups. The correlation between AL, corneal curvature, intraocular pressure and choroidal thickness was analyzed by Pearson correlation. ResultsThe choroidal thickness in the foveal macula and the areas 1 mm and 3 mm away from the fovea were compared among the control group, HM group, OHM group and SHM group, the difference were significant (χ2=76.646, 36.715, 27.660, 35.301, 24.346, 38.093, 36.275, 33.584, 36.050; P＜0.05). Compared with the control group, the choroidal thickness of the fovea and the choroidal thickness in each area within 1 and 3 mm from the fovea in the HM group, the OHM group and the SHM group were significantly thinner than those in the control group, and the difference was statistically significant (P＜0.05). There were statistically significant differences in choroidal thickness in each region between the group and the SHM group, and between the OHM group and the SHM group (P＜0.05). The results of correlation analysis showed that AL was negatively correlated with choroidal thickness in various regions (P＜0.05); SE was positively correlated with choroidal thickness in various regions (P＜0.05); corneal curvature and intraocular pressure had no significant correlation with choroidal thickness in various regions (P＞0.05). ConclusionsThe choroidal thickness of SHM is significantly lower than that of OHM and HM; OHM patients have lower SE. However, the choroidal thickness is significantly thinner. AL and SE are the influencing factors of choroidal thickness.
ObjectiveTo preliminarily investigate the mechanism of MMP-9 blocking CD73 detachment from RPE cells surface and preventing and treating experimental autoimmune pigment membranitis (EAU).MethodsRPE cells isolated from wild-type C57BL/6 and CD73 gene knockout (CD73-/-) mice were cultured in vitro, and treated with lipopolysaccharide and TNF-α to induce CD73 detachment from RPE surface. According to whether MMP-9 inhibitor CTK8G1150 was added at the same time (the final concentration was 5.0 mol/L) or not, RPE cells cultured in the two types of mice were respectively set as MMP-9 inhibitor intervention group and non-intervention control group. The cells in each group were treated with the intervention of a solvent, 1 μmol/L adenosine monophosphate (AMP), 1 μmol/L AMP, and 3 μmol/L 5' -α,β-methylene adenosine diphosphate (APCP) (AMP+APCP). The stimulating effect of RPE cells in different groups on CD4+ T cell proliferation was detected by tritiated thymidine incorporation. Adoptive immune induced EAU in wild-type B6 mice and CD73-/- mice, respectively. The receptor mice were randomly divided into the MMP-9 inhibitor intervention group and the non-intervention control group, and CTK8G1150 or the solvent were injected into the subretinal cavity 4, 7 and 10 days after adoptive immunity. CD73 mRNA and protein expression in RPE cells of recipient mice were detected by real-time quantitative PCR (RT-PCR) and Western blot. One-way ANOVA was used to analyze all experimental data.ResultsWhen the stimulation mode was AMP, the proliferation of CD4+ T cells in the C57BL/6 MMP-9 inhibitor intervention group decreased significantly compared with the non-intervention group (F=13.28, P＜0.01). When the stimulation mode was solvent and AMP+APCP, there was no statistically significant difference in the proliferation capacity of CD4+ T cells between the two groups (F=7.78, 6.58; P＞0.05). There was no statistically significant difference in the proliferation capacity of CD4+ T cells between the CD73-/- MMP-9 inhibitor intervention group and the non-intervention group (F=5.24, 6.12, 7.04; P＞0.05). RT-PCR results showed that there was no statistically significant difference in the relative expression of CD73 mRNA in RPE cells between the MMP-9 inhibitor group and the non-intervention control group (F=6.54, P>0.05). Western blot results showed that the expression of CD73 protein in RPE cells in the MMP-9 inhibitor group of B6 receptor mice was significantly increased compared with the control group (F=15.24, P＜0.01).ConclusionMMP-9 inhibitor blocks CD73 detachment from RPE cells surface and has a protective effect on EAU.
ObjectiveTo observe and analyze the superficial retinal blood flow density and its related influencing factors in the macular area of adolescents with different types of non-pathological high myopia (HM). MethodsA retrospective clinical study. From March to August 2022, 117 eyes of 117 adolescents who were admitted to Liaocheng Aier Eye Hospital due to myopia were included in the study. According to equivalent spherical degree (SE) and corneal curvature, subjects were divided into mild myopia or emmetropia group (control group), HM group, occult HM (OHM) group, and super HM (SHM) group, with 30 eyes, 28 eyes, 35 eyes, and 24 eyes, respectively. All subjects underwent medical optometry, intraocular pressure, optical coherence tomography (OCT), OCT angiography (OCTA), axial length (AL) and corneal curvature measurements. The diopter was SE. OCTA instrument was used to scan the macular region in the range of 6 mm×6 mm, and the software automatically divided it into three concentric circles centered on the fovea of the macular, namely, the central area with a diameter of 1 mm, the inner ring area with a diameter of 1-3 mm, and the outer ring area with a diameter of 3-6 mm. The superficial retinal vascular density (SRVD), vascular perfusion density (SBPD), the area, perimeter (PERIM), avascular index (AI) of foveal avascular area (FAZ) and retinal thickness were measured in the macular region as a whole and in different regions. One-way analysis of variance was used to compare the data among groups, and the least significant difference t-test was used to compare the data among groups. The correlation of AL, corneal curvature and intraocular pressure with SRVD and SBPD in macula was analyzed by Pearson correlation analysis. ResultsThere were significant differences in SRVD and SBPD in the central, inner and outer regions of macula in control group, HM group, OHM group and SHM group (P＜0.05). There were statistically significant differences in the thickness of the retina above, below and on the temporal side of the central and outer ring regions (P＜0.05). However, no statistically significant difference was in the thickness of the retina on the nasal side (P＞0.05). There was no significant difference in PERIM (P＞0.05). There were significant differences in FAZ area and AI (P＜0.05). Correlation analysis showed that AL was negatively correlated with SRVD and SBPD in macular whole and central, inner and outer ring regions (P＜0.05). Corneal curvature and SE were positively correlated with the SRVD and SBPD of macular whole, central area and outer ring area (P＜0.05). AL was negatively correlated with retinal thickness in the outer ring region (P＜0.05). SE was positively correlated with the thickness of the retina above, below and temporally in the outer ring region (P＜0.05). AL was negatively correlated with FAZ area and AI (P＜0.05). SE was positively correlated with FAZ area and PERIM (P＜0.05). Retinal thickness was positively correlated with SRVD and SBPD (P＜0.05). ConclusionsThe SRVD and SBPD of different types of HM in adolescents decreases to different degrees. The thickness of the retina in the central region is thicker, and the retina in the outer ring region is thinner. With the decrease of SRVD, the retinal thickness gradually is thinner.
ObjectiveTo investigate the effects of migration and expression from chemokine receptor 4 (chemokine receptor-4, CXCR4) of rat bone marrow mesenchymal stem cells (BMSCs) which were pretreated by atorvastatin (ATV) in vitro.MethodsIsolated, cultivated, identified the BMSCs, pretreated P4-P6 of BMSCs with different concentrations of ATV for 12 hours. The experimental group was divided into control group, 0.1 nM/L (group 0.1 nM), 1 nM/L (1 nM group), 10 nM/L (10 nM group), 100 nM/L (100 nM group), 1 000 nM/L (1 000 nM group). The mRNA and protein of CXCR4 were determined by real time-polymerase chain reaction and Western blot. Immunofluoreseence assay were used to detect the expression levels of CXCR4. The migration ability of BMSCs were measured by transwell chamber.ResultsImmunofluoreseence assay showed the protein level of CXCR4 of group 1 nM and 10 nM were significantly higher than the other group. RT-PCR and Western blot showed the protein and mRNA level of CXCR4 in 10 nM was higher than that in group 1 nM. The migration ability of group 10 nM was higher than 1 nM and control group.ConclusionsATV can be dose-dependent promote expression levels of CXCR4 of BMSCs cultivated in vitro.
ObjectiveTo observe the effect and mechanism of human umbilical cord blood mesenchymal stem cells-derived microvesicles (hUMSCs-MVs) on the injury of the primary rat retinal ganglion cells (RGCs) by high glucose environment. Methods The primary RGCs of Sprague Dawley rats were cultured in vitro, hUMSCs-MVs were isolated and extracted by ultra-centrifugation. hUMSCs-MVs were internalized with RGCs. The RGCs were divided into 4 groups under the conditions below: normal control group (group A), high glucose condition group (group B, RGCs+glucose 33 mmol/L), normal RGCs co-cultured with hUMSCs-MVs group (group C, RGCs+hUMSCs-MVs), and RGCs co-cultured with hUMSCs-MVs in high glucose condition group (group D, RGCs+hUMSCs-MVs+glucose 33 mmol/L). The cell activity was detected by CCK-8 test. Annexin Ⅴ/PI staining detected the cell apoptosis rate by flow cytometry. And the relative expression levels of the genes such as Bcl-2, Bax and Caspase-3 were detected by fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Statistical analysis was performed by using One-way analysis of variance and SNK-q test was used for the comparison between groups. Results The hUMSCs-MVs were extracted by ultra-centrifugation, which were characterized as single or cluster of circular membranous vesicle-like structure with diameter ranging from 100 nm to 1000 nm. The flow cytometry analysis showed that hUMSCS-MVs were highly positived by the surface markers of CD44, CD29, CD73, and CD105 whereas been poorly expressed the integrin (CD49f), HLA class Ⅱ, CD34, CD45. There were significant differences in the cell activity and the apoptosis rate among 4 groups, the cell apoptosis rate of group B was higher significantly than that of group A and group D (F=107.92, P=0.000), the cell activity of group B was lower than that of group A and group D (F=382.11, P=0.000). The results of RT-PCR and Western blot showed that the relative mRNA (F=219.79, 339.198, 1 071.21; P=0.000, 0.000, 0.000) and protein (F=544.28, 295.79, 224.75; P=0.000, 0.000, 0.000) expression of Bcl-2, Bax, Caspase-3 and the protein expression of cleaved Capspase-3 (F=533.18, P=0.000) in group B and D were higher significantly than those in group A and C. The relative expression of Bcl-2 mRNA and protein in group B was significantly lower than that of in group D (P＜0.05). The relative expression of Bax, Caspase-3 mRNA and protein in group B was higher than that in group D (P＜0.05). The relative expression of cleaved Caspase-3 protein in group B was higher significantly than that in group D (P＜0.05). Conclusion The hUMSCs-MVs can protect the cultured rat RGCs from the damage of the high glucose condition through increasing the cell activity and reducing the apoptosis rate of RGCs by promoting the Bcl-2 expression, decreasing the expression of Bax and Caspase-3 and inhibiting the Caspase-3 into the activity form of cleaved Caspase-3.
ObjectiveTo observe the changes of macular structure and choroidal capillary blood flow density in patients with acute central serous chorioretinopathy (CSC).MethodsProspective cross-sectional study. A total of 24 eyes of 24 patients with monocular acute CSC (case group) diagnosed by clinical examination from Shanxi Eye Hospital during January and March 2018 were included in the study. The eyes (24 eyes) and contralateral eyes (24 eyes) of the patients in the case group were set to CSC group and contralateral eye group, respectively. Twenty-one eyes of 21 healthy volunteers with age and gender matching were selected as normal control group. The macular structure of the eyes were observed by OCT and OCT angiography (OCTA), and the blood vessel density of choroidal capillary layer in the circular area of the macular area with a radius of 1 mm was measured. The paired t-test was used to compare the differences in blood flow density in the choroidal capillaries between the three groups.ResultsThe results of OCT showed that the serous neuroepithelial detachment in the macular area was observed in all eyes of the CSC group, with or without RPE detachment being 20 or 4 eyes, respectively. Of the 24 eyes in the contralateral eye group, 13 eyes (54.2%) had thick choroidal RPE lesions (PPE). There was no abnormality in the retina and choroidal structure in the macular area of the normal control group. The results of OCTA showed that the blood flow density of choroidal capillaries in the CSC group, the contralateral eye group and the normal control group were 1.759±0.132, 1.924±0.463, and 1.940±0.033, respectively. Compared with the eyes of the contralateral eye group and the normal control group, the blood flow density of choroidal capillaries in the CSC group was significantly lower (t=6.611, 6.474; P=0.000, 0.000). There was no significant difference in the blood flow density of choroidal capillary layer between the contralateral eye group and the normal control group (t=1.328, P＞0.05). In the contralateral eye group, there was no significant difference in the blood flow density of choroidal capillary layer between PPE eyes and no RPE eyes (t=0.806, P＞0.05).ConclusionsThere is 54.2% of the contralateral eyes in the monocular acute CSC patients with PPE. The choroidal capillary layer blood flow density is lower than that of the contralateral and normal eyes.