Study on Therapy and Mechanisms of the Role of Tang Kang in Diabetic Rat and Endothelial Cells
|School||Xinjiang Medical University|
|Course||Biochemistry and Molecular Biology|
|Keywords||Diabetic retinopathy Oxidative stress Tang Kang Endothelial vascular cells NF-κb|
Objective: to study on the effect and mechanism of Tang Kang on retinal tissue of diabetic rats and endothelial vascular cells cultured in high glucose environment (ECV-304). Methods: 1) animal experiments: Sixty Wistar rats randomly pick ten as normal control group, another fifty rats were divided into five groups including diabetic group,normal group, tangkang high dosage, median dosage and low dosage group and DaNaKang group, were given streptozotocin (STZ) injection to induce the diabetic rats model. We detected the body weight and blood glucose of all experimental groups every week. Retinal vascular preparations were performed by trypsin digestion to observe the histopathologic alternations of retinal microvascular; The content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in serum and retinal tissue were detected by spectrophotometry; Through Western blot, RT-PCR to detect the protine expression of NF-κb p65 protein and NF-κb,IL-Iβ,Bax and VEGF expression of genes. 2) The cultured ECV-304 cells with Sugar concentration of 5.5mmol / L (normal control group), 15 mmol / L group)intervention sugar medium were cultured vascular endothelial cells after 48 h, 72 h and 96 h. MTT assay of different cell proliferation in the intervention group; thio-barbiturate determination of cell MDA (Malondialdehyde, MDA) content; use of double-dithiobis nitrobenzoic acid in the colorimetric determination of vascular endothelial cells glutathione GSH content.Result: 1) animal experiments: after one week, DM rats model showed the typical“3 more”phenomena Comparison of model groups and Tang Kang high dose and low dose groups, the above symptoms is light, Tang Kang low dose groups and with tanakan group without obvious changes. Model group and the Group earlier in rat normal rat body weight significantly reduced blood sugar significantly elevated, a significant statistical significance (P<0.01). Compared with the normal group, model group of rat retinal capillary in rendering more cell-free strips (approximately 5.33 times times the normal group). Compared with the model group, n high Tang Kangdose groups do not have cells i, pieces marked decline in the number of capillaries (P<0.01), compared with the model group, the treatment group in rat serum and retinal tissue SOD activity degrees, MDA content lower. And normal controlled group comparison, model group retinal organization in the NF-κb P65 protein expression raised (P<0.01), NF-κb and VEGF gene of expression obvious raised, to Tang Kang high dose group most significantly (P<0.01); and model group comparison all treatment group large rats retinal organization in the NF-κb P65 protein and NF-κb and VEGF gene of expression obvious cut, to Tang Kang high dose group effect significantly (P<0.01) And. 2) cell test: with the rise of glucose concentration and prolonged incubation time of the Group significantly reduced cell OD values (P<0.01), GSH and MDA content increased reduction (P<0.01), has an obvious time and concentration-dependent. And model comparison, Tang Kang serum containing the treatment group obviously inhibition of cell proliferation, different levels of reduction cells and MDA content in improving GSH content, Tang Kang evident the high dose group (P<0.01). Conclusion: 1) Tang Kang inhibit the development of histopathology in the retina of diabetic rats, including capillary cell apoptosis and acllular capillaries,and show protective role to the microcapillary in the retina of diabetic rats 2) Tang Kang significantly increased activity of SOD in serum and retinal tissue of diabetic rats, reduced content of MDA. 3) The protective effect of Tangkang to the retina of diabetic rats may be contributed to the down regulation of NF-κb P65 protein in rat retina;The down regulation of NF-κb, IL-1β,VEGF and Bax mRNA may play an important role in protecting the retina of diabetic rats by Tangkang. 4) high levels of glucose cause varying degrees of endothelial vascular cells from oxidative injury and serum containing Tang Kang can significantly reduce oxidative damage.