Astragalus Extract Experimental Study Of Cultured Rat Retinal Microvascular Endothelial Cells In Vitro High-sugar

Purpose: (1) To establish cell cultured model of retinal capillary endothelial cells(RCEC) in normal glucose and in high glucose in vitro. (2) To observe the effects of high glucose to RCEC. (2) To observe the effects of Chinese medicine extractive of Milkvetch Root for RCEC.Methods: (1) Establish normal glucose cell culture model of RCEC cells in vitro. (2) Using normal glucose culturing liquid make the third endothelial cells into suspension cell, count cells and plant them in 96 hole culture-bottles, then make DMEM culturing liquid of different glucose (5.5mmol/L,11mmol/L,22 mmol/L,33mmol/L,44mmol/L,55mmol/L). Cultured after 1～6 days in vitro, we collect cells and then research the effect of high glucose for them. (3) Put 0.4mg/ml extractive of Milkvetch Root H₂₀ in RCEC of cultured 48 hours as in vitro. And then determine the OD of living cells by MTT for another 48 hours. (4) After cultured with high glucose in vitro 48 hours, RCEC cells were collected and check the sum of apoptosis cells by Flow Cytometry. (5) After cultured with high glucose in vitro 48 hours, collect the culturing liquid and check the content of ICAM-1,SOD and NO .Results: (1) The cells in primary culture demonstrated a cobblestone morphology, focally forming contact inhibited monolayers. The cells were positive staining for factorâ…§antibody. (2) Retinal capillary endothelial cells proliferation have been restricted in various high glucose (44 mmol/L,55 mmol/L for 1 day, 33mmol/L,44 mmol/L,55 mmol/L for 2 days , 22 mmol/L,33 mmol/L,44 mmol/L,55 mmol/L for 4 and 6 days). The propotion of restriction in 55 mmol/L for 6 days is highest. (3) H₂₀ can promote RCEC proliferation. (4) After 33 mmol/L glucose for 2 days, the percent of apoptosis of RCEC in 0.4mg/ml H₂₀ is lower than control's and model's (P<0.01). (5) After 33 mmol/L glucose for 2 days, the content of ICAM-1 is lower than control's and model's and the content of SOD and NO is higher than control's and model's.Conclusion: (1) High glucose could restrain retinal capillary endothelial cells proliferation, and reduce the mount of RCEC by dose dependence. With the concentration rising and time lasting, cells have distinct changes in morphology and the proportion of restriction increase. (2) High glucose could accelerate the apoptosisof retinal capillary endothelial cells and increase the content of ICAM-1 and reduce thecontent of SOD and NO. (3) H₂₀ can protect RCEC from hurting of high glucose. (4)H₂₀ corrects abnormal levels of glucolytic metabolites and delays endothelial cellsreplication.