The Role Of CIRBP In Intimal Hyperplasia After Vascular Endothelial Injury

Objective: 1.To explore the role of CIRBP in intimal hyperplasia after vascular endothelial injury;2.To investigate the effect of CIRBP on the proliferation and migration of vascular smooth muscle cells(VSMCs);3.To determine whether CIRBP regulates VSMCs proliferation and migration through m TORC1 pathway.Methods: The experiment is divided into two parts: in vitro experiment and in vivo experiment.1.In vitro experiment: The vascular smooth muscle cells were divided into negative control group and adenovirus interference group.The total protein and RNA of the cells were extracted after 48 hours in complete culture medium.The expression of CIRBP protein and m RNA in vascular smooth muscle cells was explored by western blotting and PCR experiments;Then vascular smooth muscle cells were divided into Adv-NC、Adv-sh CIRBP and Adv-sh CIRBP +insulin groups.The phosphorylation level of m TORC1 classic substrate 4EBP1 and S6 were explored by western blotting;Ki-67 immunofluorescence staining and CCK-8 colorimetry were used to detect the cell proliferation ability,and scratch test was used to detect the cell migration ability.2.In vivo experiment: C57BL/6J male mice aged about 7weeks were fed adaptively for one week,they were exposed to light for 12 hours in the daytime and fed with food and water on time.The mice were divided into sham operation group and carotid artery endothelial injury group.The model of vascular endothelial injury was established by rubbing the left common carotid artery endothelial of mice with rough guide wire back and forth.The sham operation group did not use guide wire friction,and the rest of the operation process was the same as the carotid artery endothelial injury group.Observe the status of mice after the operation,the mice were fed food and water on time,and the blood vessel samples were taken according to the grouping 28 days later.The expression of CIRBP protein in the common carotid artery of mice was explored by western blotting test.In order to further explore the role of CIRBP,the mice were divided into sham operation+ Adv-NC group,sham operation+Adv-sh CIRBP group,injury+Adv-NC group,and injury+Adv-sh CIRBP group.Firstly,the lesion site of the mouse carotid artery was divided into groups and incubated with interfering CIRBP adenovirus and negative control adenovirus for about 30 minutes.In the sham operation group,the aforementioned adenovirus was also injected into the left common carotid artery site for the same time according to the grouping.Then,on the 7th,14 th,and 21 st days after the operation,the corresponding adenovirus was injected once through the tail vein,respectively.After 28 days,the blood vessels were taken and the hyperplasia of the mouse carotid artery intima was detected by HE staining.Immunohistochemical staining was used to detect the proliferation of proliferative endometrial cells.Results: Compared with the control group,the expression of protein and m RNA of CIRBP decreased after adenovirus interference with vascular smooth muscle cells.After interfering CIRBP,the phosphorylation level of substrate4EBP1 and S6 of m TORC1 decreased,and the proliferation and migration ability of vascular smooth muscle cells decreased.After adding insulin to restore the activity of m TORC1,the inhibitory effect of interfering CIRBP on cell proliferation and migration was reversed.The expression level of CIRBP protein increased after carotid endothelial injury in mice.After vascular endothelial injury,the intimal hyperplasia was obvious.Compared with the negative control group,after interfering the expression of CIRBP,the intimal hyperplasia after endothelial injury was improved.Conclusion: 1.The expression of CIRBP in proliferative intima increased after vascular endothelial injury,and interference with of CIRBP could inhibit the proliferation of vascular intima.2.The expression of CIRBP increased in VSMCs during the proliferative period,and inhibited the proliferation and migration of vascular smooth muscle cells after interfering the expression of CIRBP.3.CIRBP may promote the proliferation and migration of vascular smooth muscle cells by positively regulating the activity of m TORC1.