| Objectives To investigate the effect and mechanism of sea-buckthorn extract on retinal ganglion cells in diabetic ratsMethods Select 60 healthy male SD rats without anterior segment and fundus diseases,radom choose 40 rats of them in order to establish the diabetes model by intraperitoneal injection of streptozotocin.After the model was successfully established and the fasting blood glucose was stable,divided into diabetes group(group D)and sea-buckthorn extract treatment group(group H)randomly,with 20 rats in each group.The other rats were intraperitoneally injected with 0.01mol/L sodium citrate buffer as control group(group C).The rats in group H were given sea-thorn extract(2g/kg)by intragastric administration,and the other rats were given equal volume of normal saline instead for 12 weeks.Fasting blood glucose of all rats was measured at 1,4,8 and 12 weeks.At the end of 12 th week,exclude death and have significant difference in weight rats randomly selected 12 rats from each group radomly to continue the experiment.The rats were anesthetized by intraperitoneal injection of 2ml 4% chloral hydrate in order to get the specimen.Observing the morphology of retina by HE staining,observe the apoptosis of ganglion cells in rats by TUNEL method and calculate the apoptosis index(AI).Measuring Caspase-3 expression in each group by immunohistochemistry.Observing the relative expression levels of glial cell derived neurotrophic factor and p38 MAPKm RNA by q PCR.Results 1 Fasting blood glucose: At the same time node,diabetic group had the highest fasting blood glucose,followed by treatment group,and normal group had the lowest fasting blood glucose.The differences among the three groups were statistically significant(P<0.05).2 HE staining: Compared with normal group,the retina in diabetic group became thinner,with disordered arrangement of each layer,and the number of ganglion cell layer cells decreased.Treatment group also can be observed in the typical structure of diabetic retinal pathological changes,but the light in diabetes group obviously,retinal thickness is thicken,each layer structure is nestly arranged,increasing the numbers of ganglion cells,the visible part of the cell in vacuoles.All of these prove the seabuckthorn extract lavage treatment can delay the progress of the diabetic rats retina pathological change.3 TUNEL and AI values: Ganglion cell apoptosis was observed in the retina of rats in each group,and the AI values in the diabetic group were the highest,the treatment group was lower than the diabetic group,and the normal group was the lowest.The differences among the three groups were statistically significant(P<0.05).So confirmed that Seabuckthorn extract could reduce RGCs apoptosis.4 Caspase-3 expression:The average optical density value(AOD)of each group was calculated and compared.AOD value of the diabetic group was higher than that of the other two groups,and AOD value of the normal group was the lowest.The difference of AOD value among the three groups was statistically significant(P<0.05).Proving that sea-thorn extract could inhibit the expression of Caspase-3.5 Relative expression levels of GDNF and p38 MAPKm RNA:The relative expression level of GDNFm RNA in the diabetic group was the lowest,while that in the treatment group was lower than that in the normal group.The difference among the three groups was statistically significant(P<0.05).The relative expression level of p38 MAPKm RNA in the diabetic group was the highest and that in the treatment group was higher than that in the normal group,with statistical significance(P<0.05).It was confirmed that sea-buckthorn extract could promote the expression of GDNF and inhibit the expression of p38 MAPK.Conclusions 1 Sea-buckthorn extract can protect retinal ganglion cells in diabetic rats to a certain extent.2 The protective effect of sea-buckthorn extract may be related to p38 MAPK signaling pathway.Figure10;Table13;Reference 160... |
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