| PurposeDiscuss the influence of thrombin to IPE cells. We will evaluate that thrombin stimulted the proliferation of cultured IPE cells,then this finding provides an experimental basis for understanding the regeneration of the lens from IPE cells.Method1,I vitro rabbit IPE cells are cultured.2,Carries on the immunity histochemistry appraisal to cultured rabbit IPE cells.3,Using in vitro cultured IPE cells, observe the influence of different thrombin concentrations to IPE cells. IPE cells are vaccinated to 96 culture boards, each 0.2ml, and randomly divided into 5 groups to carry on the experiment(1-5 group), 1 groups for the control group,2-5groups for the experimental group, separately joins 0.1,0.5,1.0,5.0NU/ml thrombin, 37℃,5%CO2 incubator to culture 48 hours. Determines the OD value of the cell quantity in various cultured boards with the MTT assay.4,According to the previous experimental result, designated 0.5NU/ml is the concentration of thrombin in the experimental group. Adjust the concentration of cultured cells, then the cells are vaccinated to 96 culture boards, each 0.2ml, and randomly divided into two groups to carry on the experiment. The control group joins 0.5NU/ml thrombin, the experimental group joins 0.5NU/ml thrombin and isometric thrombin inhibitor. Put it in 37℃,5%CO2 incubator and culture for 48 hours. Determines the OD value of the cell quantity in various cultured boards with the MTT assay.5,Statistical data analysis: using SPSS11.0 software.Using the unpaired Student's t test,the different was significant as to 0.05.Result1,Primary cultured IPE cell nucleus are round, the endochylema has been filled with the black pigment grains. Cells are the polygonal or square in the crowded place and spindle-shaped in the sparse place. After handing down from generation to generation the cellular form is similar with primary cultured cells, but along with the augmented times of handing down from generation to generation, the volume of IPE cells have increased, the number of black pigment grains in the endochylema is fewer and fewer. The cell gradually becomes old and the boundary is fuzzy.2,The result of cellulars immunohistochemistry: Nearly all the endochylema present the yellowish brown color, the cell nucleus are purple.3,Different concentrations of thrombin demonstrated to the IPE cells function MTT assay result that, the thrombin has the proliferation function to in vitro cultured IPE cells, with the obvious dosage dependence. The comparision of each group and the control group has statistics significance, and along with the thrombin increasing, the proliferation function of IPE cells also increases.4,Thrombin and its inhibitor demonstrated to the IPE cells function MTT assay result that, the proliferation function of IPE cells in vitro by thrombin can be suppressed by thrombin inhibitor, and the difference has statistics significance.ConclusionThe thrombin is one of for serine proteinase super family members. Activity thrombin in the blood plasma is originally formed by way of catalyzing the decomposing the hemoglutination enzymeoriginal enzyme compound (prothrombinase complex) (thrombin factor X a and the thrombin factor Va union forms 1: 1 compound). The present research already discovered that thrombin has the function of the participation blood coagulation as well as the thrombus,and also has other functions, for example: the inflammation response, the wound healing, organization reconstruction, activating growth factor, embryogeny, as well abnormal and unusual growth regulation. The majority of thrombin exist in blood in the precursor form, namely thrombin precurosor. Pull the blood ingredient in the wound gathering to influence the healing response.In vitro cultured IPE cells have the obvious proliferated under the function of thrombin, moreover the cell proliferation has the certain dosage dependence to the thrombin. Along with the concentration of thrombin enhanced, the proliferation function of IPE cells also strengthen. In the cataract surgery, the surgery stimulates the permeability of blood vessel as well as the blood - aqueous humor barrier destruction or weaken in the surgery process all possibly causes thrombin in the blood vessel originally enter aqueous humor, and participate promotion IPE cells proliferation. After joining the thrombin inhibitor, the growth of IPE cells is suppressed. Obviously, the function of thrombin can be suppressed by thrombin inhibitor. We should see that although the inhibitory action of thrombin inhibitor has statistics significance, but it cannot be completely suppressed, in other words, the proliferative function of thrombin to the IPE cells is not only itself directaction, there possibly also has its derivative or other functions, these extra functions at present have not been understood, still waiting for the further research.Thrombin stimulted the proliferation of cultured IPE cells.This finding provides an experimental basis for understanding the regeneration of the lens from IPE cells. |
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