Study On The Protective Effect Of Buqing Granules In The VEGF-VEGFR2-SRC Pathway On The Oxidative Stress In Rat Lens

Object Objective to investigate the effect of supplementing green Granule on the balance of oxidative stress and the expression of VEGF-VEGFR2-SRC signaling pathway in rats.In order to provide experimental basis for clinical treatment of age-related cataract(ARC).Methods(1)SPF class male SD rats were examined by slit lamp microscopy to exclude bilateral lens disease and other ocular diseases.After one week adaptive feeding,in accordance with the principle of random distribution,divided into six groups: control group,model control group,Qi group,BQ granules,high,medium and low dose group,except control group,other groups were using ultraviolet irradiation copy lens oxidative stress in the rat model of SD,during the modeling at the same time give the corresponding aqueous solution by gavage.Positive control group,QJ 1g/ml solution gavage;BQ high,medium and low dose groups were given 4g/ml,2g/ml,1g/ml aqueous solution gavage;model control group and blank control group were treated with normal saline(10ml/kg)instead of medicine.The rats were observed and recorded on the general situation of the mental state,diet and drinking water during the experiment.To observe the degree of opacity of lens after UV irradiation.The degree of turbidity of the lens was recorded on the 5rd day,10 days,and 15 days after UV irradiation.Successful indicators : slit lamp observed under the rat lens are at least vacuoles.(2)After 15 days,HE staining was used to observe the pathological changes of lens epithelial cells;The activity of SOD and GSH-PX in serum and the content of MDA were measured by enzyme-linked immunosorbent assay(ELISA).Real Time PCR method was used to detect the expression of VEGF,VEGFR2,SRC in the lens of blank group,model group,positive control group and high dose group.The expression of VEGF,VEGFR2,SRC and P-SRC in lens epithelial cells was detected by immunohistochemistry.Determination of VEGF,VEGF-R2,SRC and P-SRC protein expression in lens by Western Blot.Attention: Each group of the same rat two different lenses were measured different indicators,so as not to affect the experimental results.Result(1)The rats in each group were in good condition during UV irradiation,Individual rats appear apathetic,epistaxis and other phenomena;later rats in each group were mentally retarded,eyes blurred,lethargy and other cataract performance.Under the slit lamp,the lens of blank control group were always transparent;Five days after UV irradiation,lens opacity was not obvious;On the tenth day,the fibrous structure of the lens was disordered and the edema was obvious.Most of the cortical areas of the lens were dense;Day 15,lens opacity,cataracts continue to develop.(2)During the modeling,slit lamp observation results showed that : at the beginning of the experiment,in the model group,the incidence of cataract was 57%.After three days,compared with the blank control group,the difference was significant(P < 0.05);with the prolongation of UV irradiation,the number of SD rats in stage ?,? and ? increased gradually,and there was significant difference compared with blank control group(P<0.05).The result showed that the positive control group and the treatment group can delay the development of cataract,compared with the model group,the difference was statistically significant(P<0.05),and the effect of BQ Granule high dose group was more significant(P<0.05).(3)HE: Blank control group—The lens epithelial cells were a single layer of cuboidal epithelial cells.Under microscope,the morphology of cells was regular and the fibers were arranged closely.Model group—The lens epithelial cells were irregular,the depth of structural disorder of lens fiber.In the positive control group,the damage of lens tissue was reduced,the arrangement of LECs was regular,the staining was light,and the arrangement of lens fibers was more complete.BQ group:Lens tissue injury,lens epithelial cells arranged neatly arranged more compact,QJ complete set of lens fibers,especially in the most obvious BQ high dose group.(4)Immunohistochemistry: The positive expression of VEGF,VEGF-R2,SRC and P-SRC were significantly higher in the SD cataract lens induced by oxidative stress than that in the normal control group,The difference was statistically significant(P <0.05);After treatment,the levels of VEGF,VEGF-R2,SRC and P-SRC positive expression were significantly less than model group,the difference was statistically significant(P < 0.05).Compared with QJ group,BQ grnules are better,the difference was statistically significant(P<0.05).(5)Serum results: Compared with the blank control group,SOD and GSH-PX activities in the model group were significantly decreased,while the MDA content was significantly increased,the difference was statistically significant(P < 0.05);Comparison of the BQ Granule groupand QJ group,with the concentration of BQ Granule gradually increased,the activity of SOD and GSH-PX in serum increased significantly,and the MDA content decreased,which was statistically dependent and the difference was statistically significant(P<0.05).The result showed that: after UV irradiation,the activity of SOD and GSH-PX in serum and the content of MDA were significantly decreased,BQ granule can significantly improve the activity of SOD and GSH-PX in serum and decrease the content of MDA,and with the increase of concentration,the effect is more obvious.(6)Real Time-PCR: Compared with the blank control group,the expression levels of VEGF,VEGF-R2 and SRC in the lens of the model group were significantly higher than those in the control group(P < 0.01);Compared with the model group,the expression of VEGF,VEGF-R2 and SRC in the BQ group was significantly lower than that in the model group(P <0.01)after the intervention of the drugs in the RNA group;Compared with model group,the expression of VEGF and SRC in m RNA group was not significant in QJ group(P > 0.05);BQ dose group compared with the blank control group,the expression of three factors in RNA was significantly different(P < 0.01).(7)Western-Blot: Compared with the blank control group,the expression levels of VEGF,VEGF-R2,SRC and R-SRC were significantly increased in the model group after UV irradiation(P < 0.01);After treatment,the treatment group compared with the model group,VEGF,VEGF-R2 and SRC protein expression were decreased,the difference was significant(P < 0.01);The expression level of R-SRC protein in QJ group was not significantly different from that of modelgroup(P > 0.05);BQ high dose group R-SRC protein expression level,compared with the blank control group,the difference was not significant(P > 0.05);BQ high dose group compared with QJ group,there was significant difference(P < 0.01).Conclusion(1)Oxidative stress mechanism plays an important role in the occurrence of cataract in SD rats induced by UV;(2)BQ Granules may reduce the expression of VEGF,VEGF-R2,P-SRC and SRC in the lens by reducing the expression of VEGF,VEGF-R2 and SRC in the lens,so as to reduce the lens opacity and prevent catarac.