[Inhibition of hyperplasia of vascular smooth muscle cells in rat by the plasmid containing the short hairpin RNA of angiotensin II type 1 receptor].
OBJECTIVE
RNA interference is a new technology that inhibit effectively the expression the specific genes. The current study was designed to investigate whether the plasmid containing the short hairpin RNA (shRNA) of angiotensin II type 1 receptor (AT(1)R) can inhibit the hyperplasia of vascular smooth muscle cells in rat.


METHODS
The plasmids containing the shRNA of AT(1)R were constructed, and transfected vascular smooth muscle cell (VSMC) to detect the effect on the AT(1)R expression by RT-PCR and Western blot, observe the shape of VSMCs by the inverted phase contrast microscope, and detect the hyperplasia of VSMCs by trypan blues staining and MTT.


RESULTS
The plasmids was certified to be in the right rank. After transfecting cells, there was significant difference (P < 0.01) in the expression of AT(1)R mRNA between the plasmid transfected group (pAT(1)R-shRNA(1) 1.37 +/- 0.15; pAT(1)R-shRNA(2) 1.45 +/- 0.12) and the control group (2.09 +/- 0.26), and there was significant difference (P < 0.01) in the expression of AT(1)R protein between the gene transfected group (pAT(1)R-shRNA1 1.12 +/- 0.04; pAT(1)R-shRNA2 1.20 +/- 0.07) and the control group (3.17 +/- 0.21). It is shown that pAT(1)R-shRNA can decrease the expression of AT(1)R mRNA and protein. There was significant difference (P < 0.01) in the Cell number between the plasmid transfected adding AngII group (pAT(1)R-shRNA1 5.48 +/- 0.44; pAT(1)R-shRNA2 5.55 +/- 0.45) and the AngII control group (8.13 +/- 0.41); there was significant difference (P < 0.01) in the Ratio of light density by MTT between the plasmid transfected adding AngII group (pAT(1)R-shRNA1 0.365 +/- 0.024; pAT(1)R-shRNA2 0.307 +/- 0.025) and the control group (0.485 +/- 0.011); It is shown that that pAT(1)R-shRNA can inhibit the hyperplasia of VSMCs, and matching the result of morphology observation.


CONCLUSIONS
The plasmids containing the shRNA of AT(1)R can inhibit the expression of AT(1)R mRNA and protein in VSMCs, and inhibit the hyperplasia of VSMCs induced by AngII in rat.
Sun
Cheng-Lin
CL
Department of Vascular Surgery, The First Hospital Affiliated to China Medical University, Shenyang 110001, China.
Duan
Zhi-Quan
ZQ
Xin
Shi-Jie
SJ
Feng
Zong-Cheng
ZC
Zhang
Jing-Hong
JH
Gu
Jun
J
Zhang
Qiang
Q
D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D001011:Aorta / Q000166:cytology; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D006965:Hyperplasia; D009131:Muscle, Smooth, Vascular / Q000473:pathology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism; D010957:Plasmids; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D044140:Receptor, Angiotensin, Type 1 / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D044140:Receptor, Angiotensin, Type 1; D000804:Angiotensin II