Drink Eight Taste Bright Eye To Sleep Deprivation Myopia Model Rats Scleral Fibroblast Proliferation

Scleral fibroblasts is main component of sclera, it constructs eyeball ectotheca with its deuterogenic collagenous fibers and other external cell ground substance, protects the wholeness of eyeball, and determines the length of eye axis. Sclera is a kind of intensive connective tissue, which is mainly made of scleral fibroblasts, thus it is one of the eye tissue closely related to myopia. Its structure, biochemistry and cell biology all play a key role in the development of myopia, for example, the fragile and stretchable sclera is the direct reason for pathologic myopia.GF is a kind of polypeptide substance regulating the growth and multiplication of cells. They are information elements leading to cell multiplication, which play an important role in the process of cell growth, differentiation regulation and signal transmission. The study on FDM proves that bFGF in extra-cell ground substance, TGF-β, NGF and IGF-I all play an important role in the growth of scleral fibroblasts.However, the previous experiments with fibroblasts in animals often utilized chickens or guinea pigs, instead of rats. Up to now, except reports on the influence of bFGF on rat scleral fibroblasts cultivated extracorporeal, there’s few report on the influence of TGF-β、NGF、IGF-I on rat scleral fibroblasts. Therefore, further study is indispensable.Ba Wei Ming Mu Yin is an experienced prescription based on years’clinical practice by ophthalmology of No.2Hospital Affiliated to Liaoning university of Traditional Chinese Medicine, main prescriptions are shengdi, huangqi, danggui, shanyurou, sanshen, fuzi, rougui, wuweizi. The prescription has made great effects on the treatment of pathologic myopia, though its mechanism is not revealed.Purpose By means of intracorporal animal experiment and extracorporeal rat scleral fibroblasts experiments, the study is to observe the variations of GF in bFGF、 TGF-β、NGF、IGF-I and four concentrations of GF in rat scleral fibroblasts proliferation after rats were deprived of scleral tissue, in order to explore the role of scleral fibroblasts in the process of FDM. The thesis is to observe the functions of Ba Wei Ming Mu Yin on scleral fibroblasts, discuss its mechanism, and lay a good foundation for clinical practice.Material and Method13521-day-old SD rats are selected, whether male or female.135SD rats are randomly divided into9groups,15in each group. Group Al is model8-week group; group A2is blank8-week group; group Bl is model6-week group; group B2is blank6-week group; group Cl is model4-week group; group C2is blank4-week group; group D1is model one-week group; group D2is blank one-week group; group E is Ba Wei Ming Mu Yin group. At the beginning, let the rats in group A1, group B1, group C1, group D1, group E have experiment eyelid suture, making them visual deprived. The next day after the operation, Group E rats receive decoction gavage treatment of the Chinese medicine Ba Wei Ming Mu Yin. Rats in group Al and group E experience form visual deprivation for8week; rats in group B1experience form visual deprivation for6week, rats in group C1experience form visual deprivation for4week, rats in group D1experience form visual deprivation for1week. Nothing was done about the blank groups. The rats were sacrificed at each time point measurement of the ipsilateral eye axial length, and detect the expression levels of bFGF, TGF-β, NGF, IGF-I by means of SP and RT-PCR.Medicated serum was prepared by the rats were given by gavage, intragastrically for10days,1hour after the last administration, take arterial blood serum and separate the blood, different concentrations of the drug-containing serum on in vitro cultured rat sclerafibroblasts, MTT assay was used to determine the proliferative activity of cells in each group, measured by flow cytometry groups scleral fibroblasts of the cell cycle. Results1. The axial length of one week model rats do not significantly change after4weeks, while there is a significant increase in axial length compared with the control group. Compared with model8-week group and model6-week group, the axial length of treatment group is shorter.2. Immunohistochemistry chemical experimental results:bFGF, TGF-beta, of NGF, of IGF-the Ⅰ protein expression level descends with the prolongation of the eyelid suture time, the expression level of the treatment group of bFGF, TGF-beta, of NGF, of IGF-the Ⅰ is significantly higher than the model group of eight weeks.3. RT-PCR experimental results:the expression level of bFGF and TGF-beta NGF, IGF-Ⅰ mRNA decreases with the extension of the eyelid suture time, and the expression level of treatment group of bFGF, TGF-beta NGF and IGF-Ⅰ mRNA was significantly higher than the model8-week group.4. It is proved in MTT that OD value of various concentrations of growth factor group were significantly higher than that of the blank control wells (p <0.05) was statistically significant.5. By means of flow cytometry, it is proved that compared with the blank wells of b-FGF, TGF-beta, NGF concentration intervention, cells in the G1phase was significantly reduced (p<0.05), G2and M phase cells was significantly increased (p<0.05).6. Flow cytometry proved that IGF-Ⅰ at different concentrations after the intervention, in the G1phase cells were significantly reduced (p<0.05), while cells in S phase was significantly increased (p<0.05).7. It is proved in MTT that10%,20%medicated serum hole OD values are significantly higher than that of blank groups (p<0.05).8. Compared with blank groups, the number of various concentrations of mediated serum hole cells in G1phase significantly reduces, while in S phase greatly increases (p<0.05). Conclusions1. Rats visual deprivation can lower the level of bFGF, TGF-β, NGF, IGF-I in scleral tissue, lengthen eye axis. The level is in inverse proportional to time.2. Ba Wei Ming Mu Yin can effectively control the growing length of rat model axis oculi, whose mechanism may be that, Ba Wei Ming Mu Yin c can promote the expression of bFGF、TGF-β、NGF、IGF-I, thus prevent axis oculi from lengthening.3. bFGF、TGF-β、NGF can promote the multiplication of scleral fibroblasts in dose-effect relationship, facilitating cells into phase G2and M. IGF-I can promote the multiplication of scleral fibroblasts in dose-effect relationship facilitating cells into phase S.4. Ba Wei Ming Mu Yin medicated serum can regulate the cycle of rat scleral fibroblasts, promoting cell multiplication. By driving phase G1, G2, M and S, Ba Wei Ming Mu Yin can furtherthe synthesis of DNA and protein.