| Objective: To observe the expression of Long non-coding RNA MIAT in rat with diabetic retinopathy by human umbilical cord mesenchymal stem cells intervention and the changes of retinal inflammatory factors and microvascular;To explore the role of MIAT in diabetic retinopathy after the injection of HUMSCs.Methods: Ninety healthy male Sprague-Dawley rats were randomly divided into normal control group(18)and diabetes mellitus(DM)group(72).The rats in the normal control group were randomly divided into group A(5)and group B(13),which were sacrificed at the beginning and end of the experiment,respectively.In DM group,rats were induced into DM rat model by injecting streptozotocin(STZ)via the lower left abdominal cavity.Body weight and blood sugar were monitored regularly.High sugar and high fat feeding continue.At DM 3rd,4th,5th month(m),5 rats randomly selected,which were classified as DM 3m group(5),DM 4m group(5),DM 5m group(5).Corresponding Rats were sacrificed in the month,and retinal tissue was taken.When the model was successfully established for 5m,10 rats died.The remaining rats(47)were randomly divided into DM 6m group(16),DM 6m + HUMSCs group(15),DM 6m + PBS group(16).The DM 6m+HUMSCs group was injected with 1ml of HUMSCs via tail vein,and rats in the DM 6m + PBS group were injected with an equal volume of PBS solution.The rats were killed after one month.Real-time fluorescence quantitative PCR was used to detect the expression of MIAT,IL-1? m RNA and IL-6 m RNA in retinas of each group.ELISA was used to detect the expression of IL-1? and IL-6 in retinas.The expression of Occludin in retinas was detected by immunohistochemistry.Evans blue(EB)retinal perfusion was used to detect retinal microvascular changes.Results: 1.MIAT expression in rat retina: MIAT expression in DM 3m group increased,with no significant difference compared with normal control group A(P>0.05);MIAT expression increased in DM 4m group,DM 5m group and DM 6m group,and compared with normal control group A,the difference was statistically significant(P<0.05).There was no significant difference in MIAT expression between adjacent months in DM 3m,DM 4m,DM 5m and DM 6m groups(P>0.05),while there was significant difference in MIAT expression between non-adjacent months(P<0.05).MIAT expression in DM 6m+HUMSCs group was lower than that in DM 6m group,the difference was statistically significant(P<0.05).2.The expression of IL-1? and IL-6 in retina and the correlation analysis with MIAT: The expression of IL-1? and IL-6 in DM 6m group increased,and compared with normal control group B,the difference was statistically significant(P<0.05).The expression of IL-1? and IL-6 in retina of DM 6m+HUMSCs group decreased,and compared with DM 6m group,the difference was statistically significant(P<0.05).MIAT is positively correlated with the expression of IL-1? and IL-6(r =0.733,0.790;P<0.05).3.The expression of Occludin detected by immunohistochemical staining: In the normal control group B,brown-yellow staining was observed in the ganglion cell layer and the inner nuclear layer of the retina,and ring-shaped staining was observed in the microvascular wall of the ganglion cell layer.In the DM 6m group,the staining of the ganglion cell layer,the inner nuclear layer and the microvascular was less than that of the normal control group B,and the staining became lighter.Retinal microvascular staining in DM 6m+HUMSCs group was deeper than the DM 6m group.Compared with DM 6m group,the IA values of Occludin protein in the DM 6m+HUMSCs group significantly increased,the difference was statistically significant(P<0.05).4.Retinal EB perfusion: Rats in normal control group B have regular retinal microvascular morphology and good course;and in DM 6m group,the microvascular is tortuous and dilated.After 1 month with tail vein injection of HUMSCs,retinal microvascular leakage in DM 6m+HUMSCs group was less than that in DM 6m group.The leakage of the DM 6m group was significantly increased,and compared with the normal control group B,the difference was statistically significant(P<0.05);the leakage of microvascular in the DM 6m+HUMSCs group was significantly reduced,and compared with the DM 6m group,the difference was statistically significant(P<0.05).Conclusion: 1.MIAT expression in retina of diabetic rats from third month to sixth month increases in a time-dependent manner.2.After tail vein injection of human umbilical-cord mesenchymal stem cells,the expression of IL-1? and IL-6 in retina of diabetic rats decreases,and retinal microangiopathy is alleviated.3.After tail vein injection of human umbilical-cord mesenchymal stem cells,MIAT expression in retina of diabetic rats decreases,and the expression of MIAT is positively correlated with that of IL-1? and IL-6.4.MIAT might play regulatory roles in reducing IL-1? and IL-6 levels and mitigating microangiopathy in the retina of diabetic rats after tail vein injection of human umbilical-cord mesenchymal stem cells?... |
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