| Backgroud:Glaucoma is the second leading causes of blindness in the world in comparision to cataract. Glaucoma is characterized by pathological high intraocular pressure or normal intraocular pressure with injury of optic nerve and visual function damage. Glaucoma is a kind of lifelong disease. Most patients see doctor so late that drug has been unable to meet the needs of target intraocular pressure. Instead, Glaucoma filtration operation is the main treatment. Trabeculectomy is most classic filtering operation of Glaucoma. However, failure of intraocular blood-aqueous barrier, inflammatory response of operation area, hyperplasia fibroblast and collagen of filtration area, and finally hypertrophic scar tissue under conjunctiva and closure of aqueous humor outflow channel lead to the failure after glaucoma filtering operation. At present, operation in conjunction with antimetabolite such as5-Fu and MMC is mostly adopted in clinical to suppress filtering bleb scarring. The effect is certainly consistent, but the side effect and complications is obvious. Therefore, its use is limited. Chinese medicine for anti-scarring treatment is usually Qufeng ChuShi Tongbi, Huoxue Tongluo Huayu and Qingre Jiedu Sanjie method, Chinese medicine and herbal extracts of Qufeng ChuShi, Huoxue Huayu, Qingre Jiedu is mostly adopted. However this kind of drug is more applied systemicly and less applied in ophthalmology in clinical. The mechanism of resistance to filtering bleb scarring has been no absolutely accurate description.Objective:To investigate the mechanism of Qufeng Chushi Huoxue Tongbi herbal extracts impacting on postoperative incision site about fibroblast proliferation and collagen synthesis, according to topical application of antirheumatic herbal extracts Tet and TP in filtration surgery on rabbit eyes.Methods:40New Zealand white rabbits were randomly divided into7groups, including:saline group, MMC group,0.2mg/mlTet group,0.4mg/mlTet group,0.2mg/mlTP group,0.4mg/mlTP group, normal group. The36abbits(72eyes) in6drug group undergo routine trabeculectomy and accept local infiltration with different drugs on incision site in2min. Take4rabbits(8eyes) in normal group as a control, without any intervention. Observe IOP and Anterior segment performance of eyes after6days and14days. Take3rabbits (6eyes) randomly from each drug group as6days group and kill them on the6th day after surgery. Take the remaining3rabbits (6eyes) from each group as14days group and kill them on the14th day after surgery. Prepare eyeball specimens inmmediately after all the of experimental rabbits were killed and fix them by10%formalin. Observe pathological section for HE staining and immunohistochemical staining through microscope after drawing materials.Results:1intraocular pressure monitoring in different periodOn the first day after operation, the intraocular pressure of every group was significantly decreased compared with preoperative cicumstances and recover to normal value gradually with2weeks. On the first day and sixth day after operation, there is significant difference between preoperative and postoperative intraocular pressure about the normal saline group (P <0.05);on the fourteenth day after operation, there is no significant difference between preoperative and postoperative intraocular pressure about the normal saline group (P<0.05); On the first day and sixth day after operation, there was apparently significant difference between preoperative and postoperative intraocular pressure about the MMC group (P<0.01);on the fourteenth day after operation, there is significant difference between preoperative and postoperative intraocular pressure about the MMC group (P<0.05); There was apparently significant difference that the intraocular pressure of high concentration Tet groups, low concentration Tet group, high concentration TP group, low concentration TP group after1day,6days and14days is lower than before respectively(P<0.01).On the first day after operation, the intraocular pressure of two concentration Tet and TP group is no different from the normal saline group and the MMC group respectively(P>0.05); On the sixth day after operation, the intraocular pressure of two concentration Tet and TP group is apparently different from the normal saline group respectively(P<0.01); The intraocular pressure of two concentration Tet and the high concentration TP group is apparently different from the MMC group respectively(P<0.01), while the low concentration TP group is different from the MMC group(P<0.05); On the fourteenth day after operation, the intraocular pressure of two concentration Tet and TP group is apparently different from the normal saline group respectively(P<0.01); The intraocular pressure of the high concentration Tet and TP group is apparently different from the MMC group respectively(P<0.01);The intraocular pressure of the low concentration Tet group is different from the MMC group (P<0.05), while the intraocular pressure of the low concentration TP group is no different from the MMC group (P>0.05);On the same time after operation,the two concentration of Tet group is no different(P>0.05);Only on the fourteenth day after operation, the two concentration of TP group is different(P<0.05).To sum up, the normal saline group’s postoperative intraocular pressure fluctuate and rebound quickly, nearly the same with preoperative cicumstances;the postoperative intraocular pressure of other groups rises slowly and smoothly, apparently lower than preoperative cicumstances. 2Anterior segment performance of eye in different periodOn the first day after operation, blebs of all rabbits’ anterior segment are transparent, high uplift and diffused. On the sixth day after operation, two concentration of Tet group, two concentration of TP group and MMC group’s blebs are transparent, moderate uplift, diffused and less obviously hyperemia, while the blebs of normal saline group are less transparent, lower uplift, relative limited and mild hyperemia; On the fourteenth day after operation, two concentration of Tet group, two concentration of TP group and MMC group blebs’ morphology is basically similar, which are transparent, moderate uplift, diffuse and less obviously hyperemia, while some of the blebs of normal saline group are slightly transparent, slightly elevated, limited and obviously hyperemia,some of the blebs of normal saline group Present fiber wrapped appearance.3Observation of complicationsBleb leakage, endophthalmitis, corneal epithelial defects and other complications were not found in groups.1case of postoperative corneal edema in first days and1case of anterior chamber hemorrhage were found within MMC group.4Pathological manifestations4.1HE staining6-days group:The normal saline group filtering bleb shows a large number of inflammatory cells,fibroblasts and neovascularization. The MMC group showed a few inflammatory cells and a small amount of fibroblasts. Compared with the normal saline group, the filtering areas show smaller fibroblasts, smaller nucleus, larger filtration gap and more loosen surrounding tissue. The high concentration of Tet group and TP group had no obvious inflammatory cells,small fibroblast cells, small nucleus, clearly visible filtering passage, loose surrounding tissue and rule arrangement collagen. The low concentration of Tet group and TP group had less inflammatory cells,small fibroblast cells, small nucleus, clearly visible filtering passage, loose surrounding tissue and rule arrangement collagen.14-days group:The normal saline group filtering bleb shows that collagen fibers arranged densely and subepithelial connective tissue arranged densely and in disorder. The MMC group shows that the collagen fiber arranged scatteredly and subepithelial connective tissue arranged loosely. The high and the low concentration of Tet and TP groups show that filtering passages are open, collagen fibers are scattered and subepithelial connective tissue are loosen.On the sixth day and the fourteenth day after operation, the fibroblast of two concentration of Tet group and two concentration of TP group’s number is far less than the normal saline group(P<0.01); On the sixth day after operation, the fibroblast of two concentration of Tet group and the high concentration of TP group’s number is far less than the MMC group(P<0.01). the low concentration of TP group’s number is less than the MMC group(P<0.05),the different concentrantion of Tet and TP group’s number is different(P<0.05); On the fourteenth day after operation, the fibroblast of two concentration of Tet group and the high concentration of TP group’s number is far less than the MMC group(P<0.01), the low concentration of TP group’s number is less than the MMC group(P<0.05), the different concentrantion of Tet group’s number is no different(P>0.05), the different concentrantion of TP group’s number is different(P<0.05).4.2Immunohistochemical staining for TGF-β16-days group:In each treatment group, TGF-β1staining positive particles of the filtration path fibroblasts cytoplasm are less than the normal saline group.The normal saline group shows dark brown particles, the treatment groups show pale brown particles.14-days group:In each treatment group, TGF-β1staining positive particles of the filtration path fibroblasts cytoplasm show pale brown particles which are broadly similar with the normal saline group.The number of the positive particles are less than the6-days groups.Conclusion:1The histomorphology changes of the operation area is just like postoperative glaucoma clinical manifestations and pathological changes in the normal rabbit eyes.2According to the6day after operation for early-middle stage,14days for the late stages, high concentration and low concentration of traditional Chinese medicine extract of tetrandrine and triptolide of antirheumatic can maintain relatively stable low level intraocular pressure in early-middle and late lesions. On the late stages, high concentration of TP is better than low concentration. On the same time, the effect of decreasing IOP of high concentration and low concentration of tetrandrine is better than MMC group in the early-middle and late stages. The effect of decreasing IOP of high concentration triptolide is better than MMC group in the early-middle and late stages; The effect of decreasing IOP of low concentration triptolide is better than MMC group in the early-middle stage, while in the late stage were they the same.3High concentration and low concentration of traditional Chinese medicine extract of tetrandrine and triptolide of antirheumatic in early-middle and late lesions can help maintain bleb transparent, moderate swelling, diffusion.4High concentration and low concentration of traditional Chinese medicine extract of tetrandrine and triptolide of antirheumatic can inhibit fibroblast proliferation and collagen synthesis in early-middle and late lesions, and the effect was better than MMC group. On the early-middle stage, high concentration of Tet and TP is better than low concentration respectively. On the late stages, high concentration of TP is better than low concentration.5High concentration and low concentration of traditional Chinese medicine extract of tetrandrine and triptolide of antirheumatic can inhibit the expression of TGF-β1in early-middle and late lesions. |
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