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To observe the effects of estrogen on blood pressure, ventricular remodeling and LncRNA and mRNA expression profiles in ovariectomized mice model treated with angiotensin II.
Sixty-one 6 to 7 weeks-old C57BL / 6 female mice were randomly divided into 9 groups. There were control (n=6), Sham (n=6), OVX (ovariectomy, n=6); oil (injection with oilsolution, n=6), Sham+AngII(implanted AngII micropump, n=8), OVX+AngII(n=7), Sham+ AngII+E2 (injection with E2 oil solution, n=7), OVX+AngII+E2 (n=8) and OVX+AngII+valsartan (VAL, n=7) groups. The mice were sacrificed at the end of the 6th week. The left ventricular myocardium collagen fraction (CVF%) and perivascular collagen area (PVCA)/vascular lumen area (VLA) were measured by Masson staining. The cardiac LncRNA + mRNA expression profile differences were compared among Sham, OVX, Sham + AngIIand OVX + AngIIgroups with 3 mice per group. The amplified samples verification of LncRNAs and mRNAs with the most significant fold change or clinical significance were using qRT-PCR. Immunohistochemistry and western blot were used to observe the protein expression.
1.Compared with control, sham and oil groups, SBP, left ventricular CVF % and PVCA/VAL are increased (all P < 0.05) in OVX group. After AngII, there were significant differences in SBP, DBP, left ventricular myocyte diameter, left ventricular CVF (%), PVCA/VAL among sham and OVX, sham + Ang II and OVX + Ang II groups (P < 0.05 or 0.05). After OVX+AngII,there were significant differences in the above indicators among OVX + Ang II, OVX + Ang II + E2 and OVX + Ang II+ VAL (P < 0.05).2. LncRNA and mRNA expression profiles results: Between Sham+AngII and OVX+AngII groups, there were 1109 up-regulated LncRNA ((NONMMUT030935 (Six3os1) (fold change= 58.611) was the most significant one)), 390 LncRNAs with down-regulation ((NONMMUT073340 (Gm14842) (fold change= 7.105) was the most significant one)). There were 491 up-regulated mRNA ((4930471G03Rik (fold change = 33.089) was the most significant one)) and 189 mRNAs with down-regulation ((Dbp (fold change = 8.190) was the most significant one)). 3.The expression of LncRNA Six3os1 in OVX and OVX+AngIIgroups were lower than OVX+AngII+E2, Sham+AngII and Sham groups (all P <0.05). NONMMUT024335 showed the same result but the opposite direction (P <0.05). The relative expression of Fendrr in OVX + AngII group was higher than that in other groups (P<0.05). The expression of TGF-β1 protein was significantly higher in Sham + AngIIgroup than that in the Sham group, while OVX+AngII group higher than OVX group and Sham + AngII group, OVX+AngII+E2 group lower than those of OVX+AngII(all P<0.05). Compared with Sham, Sham+AngII and OVX+AngII groups, the relative expression of STC1 in OVX group was significantly decreased, and was significantly increased in OVX+AngII group. The immunohistochemical results were also confirmed (P <0.05).The relative expression of LncRNA NONMMUT05609, which might be interacted with STC1, also showed a similar result with STC1, that was, a significant decrease in OVX+Ang II group (P <0.05).
Like valsartan, E2 would have an antagonistic effect of Ang II induced elevation of blood pressure, ventricular remodeling in OVX mice. LncRNA Six3os1, NONMMUT024335, Fendrr, STC1 protein with LncRNA NONMMUT05609 might be involved in the regulation of E2 antagonizing effect of Ang II-induced ventricular remodeling, which might play a role in regulating the expression of TGF-β1 and related pathways.