| Objective: To observe the effect of Sanxue Mingmu Tablet on the expression of P38 MAPK,MEK,PI3 K and AKT in retinal proliferative membrane by establishing a model of traumatic proliferative vitreoretinopathy in rabbits.Methods: Forty-two New Zealand white rabbits were randomly divided into A,B,C,D,E,F,and G groups,with a total of 7 groups of 6 rats.They were: group A: blank group(normal group);group B: model group;group C: Sanxue Mingmu tablets group;group D: PD98059 group;group E: LY294002 group;F: MK-2206 group;Inhibitor mixed group.30 min after modeling,the rabbits in group D,group E,group F,and group G were injected with 0.1 ml MK-2206,PD98059,LY294002 and three mixed inhibitors.Groups A,B,and C were injected intravitreally with 0.1 ml of normal saline.On the first day after operation,the Sanxue Mingmu Tablets group was administered with 10 mg solution of Sanxue Mingmu Tablets.The other groups were intragastrically administered with normal saline 10 mL/kg for 28 days.On the 1st,3rd,7th,14 th,21st and 28 th day after modeling,the anterior segment of the eye and the fundus were observed with a slit lamp and a direct ophthalmoscope.The fundus,the B-ultrasound,and the three-sided mirror recorded the fundus.After 28 days,the proliferating membrane was taken for examination.The morphological structure of each layer of retina was observed under light microscope.The expression of PI3 K,AKT,MEK and P38 MAPK protein in the retina was detected by immunohistochemistry.The relative expression of P38 MAPK,MEK,PI3 K and AKT in each group was detected by qPCR and Western Blot.Results: 1.The condition of the anterior segment of the eye: After the operation,the conjunctiva of rabbits in each model showed congestion at different degrees.Some corneas showed edema,a small amount of floating objects in the anterior chamber,and lens opacity in individual rabbit eyes.On the 7th day after surgery,the symptoms were improved and the inflammation of the anterior segment was eliminated.2.Fundus conditions: The vitreous of the model has different degrees of turbidity,and the formation of proliferating membrane and proliferation cord is obvious,which has a pulling effect on the local retina and causes retinal detachment.In the group of Sanxue Mingmu tablets,there was no obvious proliferative cord in the fundus,and there was no retinal detachment.3.The retinal tissue structure was observed under light microscope: the retinal edema and the disordered structure of each layer in the model group showed hyperplastic tissue,a large amount of inflammatory cell infiltration and collagen fiber formation.The structure of the retina in the Sanxue Mingmu group was generally normal,and a small amount of inflammatory cells were observed,and no obvious hyperplasia was observed.4.Immunohistochemistry was used to observe the expression of each index in the tissues: brown and yellow particles were found in all layers of the model group,and the expression was increased in the retinal pigment epithelial layer,optic nerve fiber layer,ganglion cells and hyperplastic tissues.The expression of Sanxue Mingmu Tablets was weak,and it was mostly expressed in the retinal pigment epithelial layer and ganglion cell layer.5.Western blot analysis of the expression of the index protein in the tissue: after 28 days of treatment,the protein content of each treatment group was reduced compared with the model group.PI3 K protein expression: There was no statistically significant difference between the blank group and the inhibitor group(P>0.05),which was different from other factors(P<0.05 or P<0.01).PI3 K protein was abundantly expressed in the model group.Compared with the model group,the expression of PI3 K and the four groups of inhibitors was significantly weaker than that of the model group,and the difference was statistically significant(P<0.01).In the drug intervention group,the combination of Sanxue Mingmu Tablets and inhibitors was not statistically significant(P>0.05).The expression of Sanxue Mingmu Tablets was lower than that of the other three groups of inhibitors,and the difference was statistically significant(P<0.01).The expression of AKT protein was similar in the combination of the combination group and the blank group(P>0.05).In the model group,the expression of histones in each drug intervention group was lower than that in the model group,which was statistically significant(P<0.05 or P<0.01).There was no significant difference between the Sanxue Mingmu Tablets group and the inhibitor mixed group(P>0.05).Compared with the other three groups of inhibitors,the expression of Sanxue Mingmu Tablets group was significantly lower than that of the three groups(P<0.05 or P<0.01).In the expression of MEK protein,the content of MEK protein in each model group was significantly increased,and the expression content in the model group was the highest,and the difference was statistically significant(P<0.01).There were statistically significant differences between the models and the model group(P<0.01 or P<0.05).In the drug intervention group,the expression of the combination of Sanxue Mingmu Tablets and the inhibitor group and PD98059 group was high,the difference was statistically significant(P<0.05),and the expression of Sanxue Mingmu Tablets group was significantly lower than that of LY294002 group and MK2206.The differences were statistically significant(P<0.01).In the expression of P38 MAPK protein,the statistical significance of each model was compared with the blank group(P<0.01 or P<0.05).Compared with the model group,the expression of each group in the model group was lower than that in the model group,and the difference was statistically significant(P<0.01 or P<0.05).In each drug intervention group,four groups of inhibitors and scattered blood The expression of the film group was higher than that of the Sanxue Mingmu group,which was statistically significant(P<0.01 or P<0.05).6.qPCR was used to detect the expression of each index in the tissues: PI3 K mRNA expression: There was no significant difference between the inhibitor mixed group and the blank group(P>0.05),and the other six groups were different from the blank group,which was statistically significant(P>0.05 or <0.01).In the model group,the expression levels of the drug intervention group were lower than those of the model group,and the difference was statistically significant(P<0.01).In the drug intervention group,the expression of the Sanxue Mingmu Tablets group was similar to that of the inhibitor combination group,and there was no statistical significance(P>0.05).The difference between the Yu group and the Sanxue Mingmu group was statistically significant(P<0.05 or P<0.01).AKT mRNA expression: The expression of AKT mRNA in each model group was increased to different extents.There was no significant difference between the group of Sanxue Mingmu Tablets and the inhibitor group(P>0.05).In the model group,the expression of each drug intervention group was significantly lower than that of the model group(P<0.01 or P<0.05).Among the drug intervention groups,there was no significant difference between the Sanxue Mingmu Tablets group and the inhibitor mixed group(P>0.05).Compared with other groups,the expression of Sanxue Mingmu Tablets was statistically significant(P<0.01 or P<0.05).MEK mRNA expression: The difference between the model group and the blank group was statistically significant(P<0.05 or P<0.01).In each model group,the model group expressed MEK mRNA significantly,and the other intervention groups and model group.In comparison,it was statistically significant(P<0.05 or P<0.01).In the drug intervention group,the expression of the combination of the Sanxue Mingmu Tablets and the inhibitors was higher than that of the PD98059 group(P<0.05),and the expression level was lower than that of the intervention group(P<0.01).P38 MAPK mRNA expression: Each model has different degrees of expression,and the model group has the most expression.There were differences between the two groups and the statistical significance(P<0.05 or P<0.01).The expression levels of each group in the model group were lower than those in the model group,and there were significant differences compared with the model group(P<0.05 or P<0.01).In each drug intervention group,the expression of the Sanxue Mingmu Tablets group was lower than that of the four groups of inhibitors,and the difference was statistically significant(P<0.05 or P<0.01).Conclusion:1.Sanxue Mingmu Tablet can improve the structure of the retina in rabbit PVR,reduce inflammatory cell infiltration,reduce tissue edema,and inhibit the production of hyperplastic tissue.2.Sanxue Mingmu Tablet can effectively reduce the expression of PI3 K,AKT,MEK and P38 MAPK in rabbit retinal proliferation membrane,but the MEK effect is weak.3.The prevention and treatment of PVR by Sanxue Mingmu Tablet may be related to MAPK and PI3K/Akt signaling pathways. |
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