ABSTRACT
Objective·To investigate the effects of insulin-like peptide 6 (Insl6) on renal fibrosis and calcification in unilateral ureteral obstruction (UUO) mice. Methods·Twenty-four SPF male mice with genotypic background of C57BL/6 were divided into Sham (n=8), UUO+saline (n=8) and UUO+Insl6 (n=8) groups randomly. Mice were sacrificed 10 days after operation and renal tissues of surgical side were obtained. Sirus red staining, Masson staining and alizarin red S staining were used to verify the level of collagen and calcium deposition. TGF-β1 expression was determined by Western blotting. Realtime-PCR was used for determining TGF-β1, BMP2, Col1a1, and Col2a1 mRNA expression. Results·Compared with sham group, fibrotic area especially collagen Ⅰ , calcium deposition, TGF-β1 protein, and TGF-β1, BMP2, Col1a1, and Col2a1 mRNA expression in UUO+saline group significantly increased (all P<0.05). As compared with UUO+saline group, fibrotic area especially collagen Ⅰ, calcium deposition, TGF-β1 protein, and TGF-β1, BMP2, Col1a1, and Col2a1 mRNA expression in UUO+Insl6 group significantly decreased (all P<0.05). Conclusion·Insl6 inhibits UUO-induced renal fibrosis and calcification, which may be related to regulation of TGF-β1, collagen Ⅰ , BMP2 and collagen Ⅱ expression levels.
ABSTRACT
Objective· To prepare nanocarriers capable of improving the immunogenicity of cholesteryl ester transfer protein (CETP) hapten. Methods · Prepare CETP polypeptide modified by thiol in Fmoc, then Sulfo-SMCC were used to prepare CETP peptide-ovalbumin (OVA) nanoparticle and CETP peptide - OVA molecule conjugates in a two-step reaction scheme. The particle size and zeta potential of nanovaccine were determined and the morphology was observed. New Zealand White rabbits were vaccinated by subcutaneous injection of vaccine and serum collected from rabbits was detected by ELISA assay for the analysis of antibodies, high density lipoprotein cholesterol(HDL-C) and low density lipoprotein cholesterol (LDL-C). The rabbits were randomly allocated to the PBS group (n=3), traditional vaccine group (n=3), and nanovaccine group (n=3). Results · Nanovaccine targeting CETP were successfully synthesized, with about 70 nm in size and about -8.81 mV in zeta potential, and possessed homogeneous spherical shape under transmission electron microscopy. Compared with traditional vaccine group, rabbits in nanovaccine group got higher antibodies. Conclusion · Nanovaccine improve immunogenicity of CETP haptens, and stimulate experiment rabbits to produce higher antibodies.
ABSTRACT
The aim of the present study was to investigate the effect of ERK on 17beta-estradiol (E(2)) inhibition of vascular smooth muscle cell (VSMC) proliferation in rats after vascular injury. Common carotid artery balloon-injury (Inj) model was established in ovariectomized rats (OVX). Female SD rats were randomly divided into 4 groups: OVX, E(2)+OVX, OVX+Inj, and E(2)+OVX+Inj groups. The thickness of the vessels, the plasma content of NO, and the expression of ERK, phosphorylated ERK as well as eNOS protein were measured. The results showed that compared with OVX, the vessel wall was significantly thickened and the plasma content of NO was significantly decreased in OVX+Inj group. E(2) significantly decreased the vessel thickness but increased the plasma NO content after balloon injury. E(2) inhibited the expression of ERK, phosphorylated ERK and induced the eNOS expression. There is a positive correlation between plasma NO content and eNOS protein expression, while there is a negative correlation between plasma NO content and the thickness of vessel. The plasma NO content and the expression of ERK protein were negatively correlated. These results suggest that E(2) increases the vascular eNOS protein expression and NO release, leading to the inhibition of VSMC proliferation after balloon injury by inhibiting the ERK and phosphorylated ERK protein expression.
Subject(s)
Animals , Female , Rats , Carotid Artery, Common , Pathology , Catheterization , Cell Proliferation , Estradiol , Pharmacology , Extracellular Signal-Regulated MAP Kinases , Physiology , Muscle, Smooth, Vascular , Cell Biology , Myocytes, Smooth Muscle , Cell Biology , Physiology , Nitric Oxide , Blood , Nitric Oxide Synthase Type III , Metabolism , Ovariectomy , PhosphorylationABSTRACT
Clinical epidemiologic data and animal experimental studies regard estrogen as being protective against the development of cardiovascular diseases. The mechanisms by which estrogen affects the development of vascular diseases are not clear. Recent studies demonstrated that the cardiovascular protective effects of estrogen are closely related to nitric oxide (NO) pathway. Our previous study proved that estrogen inhibited the proliferation and oncogene expression of vascular smooth muscle cells (VSMCs) induced by endothlin 1 (ET-1) and serum,this effect was mediated by NO release. In the present study, we investigated the role of inducible nitric oxide synthase (iNOS) in the VSMCs cycle arrest induced by 17 beta-estradiol (E(2)). The effects of E(2) on iNOS activity and protein expression in cultured rat VSMCs and the influence of NOS inhibitor N(G)-nitro-L-arginine methylester (L-NAME) on the inhibitory effect of E(2) on cell cycle were investigated. NOS assay kit was used to measure the activity of iNOS and protein expression of iNOS was determined by Western-blot. Cell cycle analysis was accessed by flow cytometry. The results obtained showed that E(2) increased iNOS activity of VSMCs but not in a dose-dependent manner. E(2) 10 nmol/L increased the iNOS activity of VSMCs distinctly at two time points: 30 min and 12 h. These effects were significantly inhibited by estrogen receptor (ER) antagonist Tamoxifen (0.1 micromol/L) and NOS inhibitor L-NAME (1 micromol/L). E(2) increased iNOS protein expression of VSMCs in a dose-dependent manner. The effect of E(2) on iNOS protein expression of VSMCs started at 3 h, distinctly increased at 12 h and then decreased. Tamoxifen significantly inhibited the E(2)-induced iNOS protein expression of VSMCs. ET-1 increased cell percentage of S phase and G(2)+S/G(1). This effect was inhibited by E(2). L-NAME significantly attenuated the inhibitory effect of E(2) on cell cycle of VSMCs. The results suggest that E(2) induced G(1) arrest of VSMCs, which was associated with an increase in iNOS activity and protein expression of VSMCs. These effects were at least mediated by estrogen receptor partly.