Validation of a preclinical dry eye model in New Zealand white rabbits during and following topical instillation of 1% ophthalmic atropine sulfate

Background : The objective of this study was to validate an animal model for dry eye during and after the administration of 1% ophthalmic atropine sulfate(OAS) in New Zealand white(NZW) rabbits.Methods : OAS(1%) was applied three times per day to 30 eyes of 15 healthy NZW rabbits. Sacrifice, enucleation, and lacrimal gland removal took place on days 15, 21,and 30(OAS group). A second group(n = 5) was used as control. Clinical evaluations took place on days 3, 10, 15, 18, 21, 24 and 30. The primary endpoints were:Schirmer I test, tear break-up time(TBUT), and corneal fluorescein staining. As secondary endpoints, clinical changes including intraocular pressure, and histopathology were evaluated.Results : While OAS was administered, the Schirmer I test showed a statistically significant reduction for OAS group versus control( p < 0.001), and versus basal production( p < 0.001). TBUT showed statistically significant differences between groups(days 3 and 10;p = 0.001) and versus basal values(day 3;p < 0.001). Fluorescein staining showed a statistically significant difference(day 3;p = 0.001). The most frequent clinical finding was conjunctival hyperemia(76.9% OAS vs. 20% control). For histopathology, all OAS subjects presented some degree of inflammation(86.7% minimal;13.3% mild) whereas the control presented only 30% minimal inflammation. Goblet cell density showed no difference.Conclusions : The effectiveness of the OAS dry eye model in NZW rabbits as reported in previous studies was confirmed, provided that the application of the drug is maintained throughout the intervention;it is not a viable model after OAS administration is suspended.

Effects of Buddlejae Flos Granules on Inflammatory Factors TGF-β1,NF-κB,IL-10 and IL-12 in Lacrimal Gland Cells of Castrated Male Rabbits

Objective To observe the effects of Buddlejae Flos (Mi Meng Hua,密蒙花) granules on the expression of TGF-β1, NF-κB, IL-10 and IL-12 in lacrimal gland cells in castrated male rabbits. Methods A total of 30 New Zealand male white rabbits were randomly divided into 5 groups. Group A: blank group;group B: model group;group C: Buddlejae Flos (Mi Meng Hua,密蒙花) granules group;group D: placebo group;group E: testosterone group. Except for the group A, all rabbits were treated with bilateral testicular and epididymis resection before intervention. Rabbits in group C were administered with Buddlejae Flos (Mi Meng Hua,密蒙花) granules (100 mg/kg), 3 times per day;Rabbits in group D were administered with normal saline, 3 times per day. Rabbits in group E were injected with testosterone propionate (0.5 mL/kg) in the thigh muscle, every 3 days. Schirmer I test (SIT) and break-up time (BUT) were measured on 1st day before modeling and 30th day after modeling. After 30 days of intervention, all rabbits were sacrificed. The removal lacrimal gland was stained in immunohistochemica staining. The expression of TGF-β1, NF-κB, IL-10 and IL-12 in lacrimal gland tissue of each group was detected, and the structure of lacrimal gland was observed. Results The results of SIT and BUT showed that there was significant difference between group B and D compared with group A (P < 0.05). There were no significant differences between group C and E (P > 0.05). There were no significant differences in the comparison of inflammatory factors TGF-β1, NF-κB, IL-10 and IL-12 between group B and D (P > 0.05). There were significant differences between group B and D compared with other groups (P < 0.01). There was no significant difference between group A, C and E (P > 0.05). Conclusions Buddlejae Flos (Mi Meng Hua,密蒙花) granules have an inhibitory effect on the expression of TGF-β1, NF-κB, IL-10 and IL-12 in castrated male rabbit lacrimal gland cells, and alleviate the inflammation of lacrimal gland tissue by inhibiting the expression of TGF-β1, NF-κB, IL-10 and IL-12 in lacrimal gland tissue, so as to treat dry eye.

Effect of Flos Buddlejae Granules on Apoptosis Factors Bax,Bcl-2,Fas and FasL in Lacrimal Gland Cells of Castrated Male Rabbits

Objective To observe the effect of different concentration Flos Buddlejae(Mi Meng Hua,密蒙花)granules on apoptosis factors of lacrimal gland cells of castrated male rabbits,and study the treatment effects of Flos Buddlejae(Mi Meng Hua,密蒙花)granules in the dry eye model of castrated male rabbits.Methods Flos Buddlejae(Mi Meng Hua,密蒙花)raw material was made into granules.Thirty healthy adult New Zealand rabbits were randomly divided into five groups,six rabbits in each group.Group A:blank group,Group B:model group,Group C:Flos Buddlejae(Mi Meng Hua,密蒙花)granule group,Group D:placebo group,Group E:testosterone group.Except for group A,all rabbits underwent removal of bilateral testis and epididymis.Rabbits in group C were administered Flos Buddlejae(Mi Meng Hua,密蒙花)granules(100 mg/kg),three times per day;rabbits in group D were administered normal saline,three times per day.Rabbits in group E were injected with testosterone propionate(0.5 mL/kg)in the thigh muscle,every 3 days.All rabbits were tested by Schirmer I test(SIT)and tear film break-up time(BUT)before operation and 4 weeks after operation.After 4 weeks,all rabbits were sacrificed by air embolism and clipping of the lacrimal gland.Apoptosis factors,including Bax,Bcl-2,Fas and FasL of lacrimal gland cells were characterized by immunohistochemistry,and resulting data were statistically analyzed.Results(1)Comparison of SIT and BUT before and after operation:There were statistically significant differences between groups B and D(P<0.01),but not among other groups(P>0.05).(2)Comparison of apoptosis factors Bax,Bcl-2 Fas and FasL:In a comparison of groups B and D,there was no statistically significant difference after operation(P>0.05).In a comparison of the other groups,there were statistically significant differences(P<0.01).In comparisons among A,C and E groups,there were no statistically significant differences(P>0.05).Conclusion Compared with androgen,Flos Buddlejae(Mi Meng Hua,密蒙花)granules caused similar but slightly weaker depression of Bax,Fas and FasL,and increased expression of Bcl-2.