The Effect and Mechanism of p38MAPK Inhibitor CBS3830 on Intimal Hyperplasia in Autogenous Vein Grafts of Diabetic Rats
|School||Anhui Medical University,|
|Keywords||vein graft p38mitogen-activated protein kinase(p38MAPK) intimal hyperplasia inflammation diabetes mellitus nuclear transcription factor kappa B (NF-κB) advanced glycation end products (AGEs)|
Objective To study the anti-proliferative effects of the p38MAPK inhibitor CBS3830 in autogenous vein graft of diabetic rats, to further explore the mechanism of the p38MAPK signaling pathway in vein graft restenosis for preventing intimal hyperplasia and vein graft stenosis to provide a new way of the targeted therapies.Methods 30 male Sprague-Dawley rats were randomly divided into three groups: control group (n=10), sham-operated group (n=10), drug group (n=10).Sham-operated group, only simulated surgical procedure,without vascular grafts and drug intervention. Diabetic rat model were established by intraperitoneal injection of streptozotocin (STZ) in control group and drug group. Autogenous vein graft model was established with the improved cuff technique in the diabetic rats.One hour before modeling, CBS3830 of 0.3mg/ml Concentration is poured into the drug group rats through stomach tube,the dose is 10ml/kg. As for the control group was given the same volume of 1% methylcellulose works as the solvents for CBS3830. Blood samples were harvested at day 1, day 3, day 7 after surgery and before surgery.Enzyme-linked immunosorbent assay (ELSIA) was used to detect the serum levels of TNF-α, IL-1βand AGEs. Vein graft samples were harvested day 7 after surgery for histological examination. HE staining was used to detect the pathological changes of vascular intima and media each group. Immunohistochemical staining was used to detect the expression of nuclear transcription factor kappa B (NF-κB) of vascular specimens each group.Western blotting was used to detect the expression of p38, p-p38,β-actin. Results The serum levels of TNF-αrise in 0 ~ 3d and reach to maximum at the third day in the drug group,showing no difference compare to control group(p>0.05). The serum levels of TNF-αshow a gradual declining trend in 3d~ 7d and reach to maximin at the seventh day, which are significantly lower than control group and show a significant statistical difference (p<0.05). The serum levels of TNF-αin sham group are significantly lower than the other groups(p<0.05). The serum levels of IL-1βreach to maximum at the first day in the drug group,then show a gradual declining trend in 1 ~ 7d ,the lowest at the seventh day and show a significant statistical difference (p<0.01). The serum levels of IL-1βin sham group are significantly lower than the other groups,too(p<0.05).The serum levels of AGEs in control group and drug group are significantly more than the sham group,showing a significant statistical difference(p<0.01),but having no difference between the two groups(p>0.05).The intima and media thickness in the drug group is significantly lower than control group, there are significant difference (p<0.05), but there is no difference compare to sham group(p>0.05).The levels of NF-KB expression of drug group were significantly lower than control group at the seventh day (p <0.05).Western blotting detection of p38 and P-p38 in the drug group is significant less than the control group, but having no significant difference compare to the sham group.Conclusion p38MAPK inhibitor CBS3830 has a significant anti-inflammatory and anti-proliferative effect in the autogenous vein graft of diabetic rats. The p38MAPK pathway activation is a critical part of vein graft vascular remodeling. It is expected a new target for preventing and treating restenosis after CABG in diabetes through further study it.