| ObjectiveTo observe the effect of Borneol on the distribution of CNTF eyedrops in the retina and the intervention of sodium iodate in SD rats,and to explore whether borneol can enhance the effect of protecting retinal nerve cells by increasing the bioavailability of CNTF eyedrops in the target organ retina,and to explain the effect and mechanism of Borneol on the menstruation Objective basis is provided for clinical application of Ophthalmology,and innovative ideas and new methods are provided for research and development of eye drops for prevention and treatment of retinal diseases.Methods1.The effect of Borneol on the distribution and metabolism of CNTF in rabbit retina:96 male New Zealand rabbits,1.5-2kg,were randomly divided into six groups:normal group,0.1%borneol matrix eye drops group,2?g/ml CNTF eye drops group,0.1%borneol+CNTF eye drops group,0.05%borneol+CNTF eye drops group,0.025%borneol+CNTF eye drops group.There were 16 rabbits in each group.In addition to the normal group(without any treatment),the rest were given 0.1%borneol matrix eye drops,2?g/ml CNTF eye drops,0.1%borneol+2?g/ml CNTF eye drops,0.05%borneol+2?g/ml CNTF eye drops,0.025%borneol+2?g/ml CNTF eye drops.The retinas of rabbits were taken out in 15mins,30mins,1h,and 2 hours after the last eye spot.The content of CNTF protein in retinas of different groups was observed by enzyme-linked immunosorbent assay(ELISA).2.Effects of borneol on the distribution and expression of CNTF eyedrops in the retina of dry age-related macular degeneration rats:a dry AMD model was established by injection of sodium iodate into tail vein.After modeling,SD rats were randomly divided into model group,borneol eye drops group,CNTF eye drops group,borneol+CNTF eye drops group,and each group had 8 rats.The rats in the right eye were given gel matrix,0.025%borneol gel eye drops,and 2?g/ml CNTF eye drops,0.025%borneol+2?g/ml CNTF eye drops,4 times/d,the administration time was 8:00,11:00,14:00 and 17:00 respectively.The normal group was fed without treatment.Immunofluorescence was used to detect the location of exogenous CNTF protein in the retina of SD rats 7 days and 14 days after administration,and reverse transcription PCR was used to detect the expression of endogenous CNTF.3.The effect of Borneol on dry eye drops of CNTF to protect retinal degeneration:the methods of making model,grouping and administration were the same as above.Electroretinogram,histopathological indexes and GFAP expression were detected 7 and 14 days after administration.Results1.Effects of Borneol on CNTF Target Distribution and Metabolism in Rabbits:At 15 min,the highest concentration was in the 0.025%borneol CNTF group(42.72±0.97pg/ml),followed by the 0.1%borneol CNTF group(39.88 ±0.71pg/ml),the 0.05%borneol CNTF group(35.84±0.55pg/ml),and the CNTF group.(35.30±1.17 pg/ml),the difference between the groups was statistically significant(P<0.05).At 30 minutes after administration,the content of CNTF in the retina of each group decreased significantly.The trend between groups was the same as that at 15 minutes.It was 0.025%borneol CNTF(18.05 ± 0.75 pg/ml)>0.1%borneol CNTF group(16.55±1.23 pg/ml).>0.05%borneol CNTF group(9.97 ± 1.42 pg/ml)>CNTF group(9.30±1.40 pg/ml)(P<0.05);after 1 hour of administration,the content of CNTF in the retina of each group continued to decrease,and 0.025%borneol CNTF group Still the highest(14.72 ± 1.09pg/ml)compared with other groups with statistical significance(P<0.05);two hours after administration,except for 0.05%borneol CNTF group content increased slightly,the other three groups content was more than the administration After 1 hour,the concentration of each group was 0.025%borneol CNTF group>0.1%borneol CNTF group>0.05%borneol CNTF group>CNTF group,the highest concentration of 0.025%borneol CNTF,the difference was statistically significant(P<0.05),the differences between the other three groups were not statistically significant(P>0.05).2.Effects of borneol on the Distribution and expression of CNTF in the retina of dry AMD Rats:Immunofluorescence test showed that there was no obvious red fluorescence in the model group at 7 days after administration.In the CNTF group,punctate red was visible except for Ganglion cell layer(GCL),inner nuclear layer(INL)and choroid layer.In addition to fluorescence,short strip-shaped red fluorescence can be seen inside the retinal pigment epithelium(RPE).In addition to the above-mentioned red fluorescence visible in the borneol CNTF group,the vitreous is near the retinal inner limiting membrane and the subretinal pigment epithelium Visible stripe fluorescence.At 14 days after administration,lump fluorescence was seen in the retinal pigment epithelium of the CNTF group,short strips of red fluorescence were seen in the space below the retinal pigment epithelium,and the red fluorescence staining of the borneol CNTF group was significantly increased compared to other groups,distributed in the retinal pigment epithelium,Outer nuclear layer(ONL),subretinal space of retinal pigment epithelium,outer plexiform layer(OPL),inner core layer.The results of RT-qPCR showed that the relative expression of endogenous CNTF mRNA in each group was normal group at 7 days after administration:1,model group:1.65 ± 0.46,borneol group:1.94 ± 1.12,CNTF group:1.62 ± 0.42,borneol CNTF group:1.72±0.92.There was no statistically significant difference between the model group,borneol group,CNTF group and borneol CNTF group(P>0.05).At 14 days of administration,the relative expression levels of endogenous CNTF mRNA in each group were:normal group:1,model group:1.84±0.92,borneol group:1.40±0.69,CNTF group:1.33±1.07,borneol CNTF group:1.23 ±0.88.There was no statistically significant difference between the model group,borneol group,CNTF group and borneol CNTF group(P>0.05).3.Effect of borneol on CNTF eye drops in protecting retinal degeneration:The ERG test results showed that the dark adaptation 3.0ERGa wave amplitude at 14 days after administration,the data of the normal group,model group,borneol group,CNTF group and borneol CNTF group were:148.06±34.70,10.61±4.52,10.61±4.52,13.37±7.00,24.30±8.46,46.48±5.65,the difference between the data of each group is statistically significant(P<0.05);The model group value is significantly lower than the normal group(P<0.05),the borneol CNTF group is significantly higher than the model group,Borneol group and CNTF group(P<0.05).The values of dark adaptive oscillatory potential(OPS)normal group,model group,borneol group,CNTF group and borneol CNTF group are respectively:134.66±42.75,10±1.62,11.26±1.5,25.61±3.04,57.41±18.64,and the data difference of each group were statistically significant.(P<0.05);The model group was significantly lower than the normal group(P<0.05),and the borneol CNTF group was significantly higher than the model,borneol and CNTF groups(P<0.05).The apparent adaptive scintillation photopotential amplitude constant group,model group,borneol group,CNTF group and borneol CNTF group are:54.66±9.80,10.50±1.97,10.40±4.46,10.70±2.16,20.14±5.05,and the differences in the data of each group are statistically significant.(P<0.05);The model group was significantly lower than the normal group(P<0.05),and the borneol CNTF group was significantly higher than the model,borneol and CNTF groups(P<0.05).In the borneol CNTF group,the left and right eyes contrast dark adaptation ERG,maximum response ERG,dark adaptation oscillation potential,bright adaptation ERG,and flickering ERG.The data of the left eye are:39.01±30.3,21.487.01,28.67±6.50,17.40±5.98,16.91±2.89,the right eye data are:58.59±40.96,46.48±5.65,57.41±18.64,16.95±6.33,20.14±5.05,among which the dark adaptation maximum response ERG amplitude,dark adaptation shock potential amplitude,and bright adaptation flicker photo potential The amplitude of the right eye was significantly higher than that of the left eye(P<0.05).Histopathological observation results showed that the retinal pigment epithelium and outer nucleus of the model control group had obvious damage,the number of cells decreased,the arrangement was disordered,and wave-like changes were observed at 7 days after administration.The normal ONL cell number group,model group,and borneol group The data of CNTF group,CNTF group and borneol CNTF group were:199.33±71.95,107.63±13.90,109.88±11.27,108.5±10.77,125.12±9.14,the model group values were significantly lower than the normal group(P<0.05),and the borneol CNTF group was significantly higher In the model group,borneol group and CNTF group(P<0.05);The change in the model group was similar to that in 7 days at 14 days of administration.The data of the normal ONL cell group,model group,borneol group,CNTF group,and borneol CNTF group were 134.66±42.75,10±1.62,11.66±1.5,25.61±3.04,57.41±18.64,the model group values were significantly lower than the normal group(P<0.05),and the borneol CNTF group was significantly higher than the model group,borneol group and CNTF group(P<0.05).The results of GFAP inununofluorescence showed that there were a few plaque-like GFAP red fluorescence staining in the ILM of the retina of normal rats,and no positive staining was found in the other layers of the retina.At 7 days,the retinal ILM in the model group showed that the GFAP red fluorescence was significantly enhanced and widened,and the strip-shaped red fluorescence penetrated from the ILM layer to the inner core layer.The borneol group is similar to the model group.The core layer and inner plexiform layer have weaker fluorescence than the model group.The CNTF group and the borneol CNTF group have no obvious red fluorescent coloration.The fluorescence of the inner boundary film of the borneol CNTF group is relatively weak.The CNTF group is slightly weaker.At 14 days,the performance trend of each group was similar to that of 7 days.The red fluorescence expression of each group was further enhanced.Very strong red fluorescence was seen in the inner limiting membrane,nerve fiber layer,and ganglion cell layer of the model group.Strip-shaped fluorescence increased,increased,and thickened throughout the retina.Stripe fluorescence was also seen in the CNTF group and the borneol CNTF group,but it was mostly limited to the inner boundary membrane to the inner plexiform layer,and the outer membrane of the borneol CNTF group to the ganglion cell layer.Less fluorescence than CNTF group.Conclusion1.0.025%borneol can effectively increase the content of CNTF protein in CNTF retina gel in rabbit retina,and may play a role through conjunctival pathway.2.Borneol can promote the distribution of exogenous CNTF in the retina of experimental dry amd rats,suggesting that borneol can increase the targeted bioavailability of CNTF by eye drops3.Borneol CNTF eye gel can significantly reduce the degree of retinopathy and improve the retinal function in experimental dry AMD rats,and its protective effect is significantly enhanced than that of CNTF eye drops.Borneol plays an important role in inducing eye drops in eye drops. |
PDF Full Text Request