Inflammation gene expression and comparison of relative composition of plaques in the control, NC and MEF 2A RNAi groups.

<p>A, B, and C: inflammatory markers (MCP-1, MMP-8 and TNF-α) in the control, NC, and MEF 2A shRNA <u>groups; D</u>, E,F and G plaque morphology in the control, NC, and MEF 2A shRNA groups; Plaque area (D), Fibirious cap thickness (E),collage content (F) and lipid content (G) were shown for the control, NC and RNAi groups. H, I and J: MCP-1 (H), MMP-8 (I) and TNF-α (J) mRNA expression in carotid plaques of the control, NC and shRNA groups at week 10 was detected by real-time PCR. Data were expressed as the mean ± SD. * <i>P</i> < 0.05.</p

Efficiency of lentviral shRNA vector transduction in the carotid plaques.

<p>A, B, C. GFP expression in sections of the carotid plaques was imaged from NC group. Carotid plaques at 1, 2 and 4 weeks following transduction were visualized under fluorescent microscope. (200×).</p

Lentiviral vector mediated knockdown of MEF 2A in mice aortic endothelial cells (MAECs).

<p>MAECs were transduced with 50 MOI of 4 different shRNA vectors and MEF 2A expression was measured on day 4 by real time RT-PCR following transduction. The GAPDH was used as an internal control. (A). MEF 2A mRNA expression detected by realtime RT-PCR (*P < 0.05) (B) MEF 2A protein expression in lentivral shRNA transduced MAECs was quantified using Image J. (C) MEF2A expression in lentiviral shRNA vector transduced MAECs was determined by Western blots. Data were presented in mean±SD. *P < 0.05. (n = 8)</p

Knockdown of MEF 2A in vivo.

<p>(A) mRNA expression of MEF 2A in the plaques of the control, NC, and MEF 2A shRNA groups at week 10; (B) Protein expression of MEF 2A in the plaques of the control, NC, and shRNA groups; (C) The concentration MEF 2A in the plasma; (D) Representative Western blots used for quantification. Data are presented in the mean ± SD. * P < 0.05</p

Body weight, plasma TC, TG, HDL-C and LDL-C levels among all groups.

<p>Data are reported as the mean± SD. <i>P</i> > 0.05 among all groups. BW = body weight; TC = total cholesterol; TG = triglyceride; HDL-C = high-density lipoprotein cholesterol; LDL-C = low-density lipoprotein cholesterol; NC = negative control group. Control = control group; RNAi = RNA interference group.</p><p>Body weight, plasma TC, TG, HDL-C and LDL-C levels among all groups.</p

Effects of Tongxinluo (TXL) on rat kidney antioxidant activities.

<p>Quantitative analysis of MnSOD (A) PCR analysis and (B)SOD activities. Quantitative analysis of catalase (C) PCR analysis and (D)activities. Data are mean±SEM; n = 10 rats per group, ^#P<0.05 and ^$P>0.05, vs. WKY group, respectively;^*P<0.05 vs. SHR group. MnSOD, manganese superoxide dismutase; PCR, polymerase chain reaction; WKY, Wistar-Kyoto; SHR, spontaneously hypertensive rat.</p

Functional evaluation of effects of Tongxinluo (TXL)in hypertensive kidney injury.

<p>Quantitative data of (A) urinary albumin excretion rate and (B)creatinine clearance. Data are expressed as mean±SEM; n = 10 rats per group, ^#P<0.05 and^*P<0.05, vs. WKY and SHR groups, respectively. WKY, Wistar-Kyoto; SHR, spontaneously hypertensive rat.</p

Effects of Tongxinluo (TXL) on rat glomerular injury in hypertensive kidney injury.

<p>Representative images of (A)PAS of kidney tissue sections. Scale bar, 50 μm. Quantitative analysis of (B) glomerular sclerosis index. (C)Representative images and (D)Quantitative analysis of desmin immunostaining. Scale bar, 20 μm. Data are expressed as mean±SEM; n = 10 rats per group, ^#P < 0.05 and^*P < 0.05 vs. WKY and SHR groups, respectively. PAS, periodic acid-Schiff; WKY, Wistar-Kyoto; SHR, spontaneously hypertensive rat.</p

Tongxinluo (TXL) altered forkhead box O1 (FoxO1) signaling in spontaneously hypertensive rat (SHR) kidneys.

<p>Western blot analysis of (A) FoxO1 and phospho-FoxO1, (B) ERK1/2 and phospho-ERK1/2, (C) p38 and phospho-p38, (D) PI3K and phosphor-PI3K, and (E) Akt and phosphor-Akt (F) AMPK and phosphor-AMPK, and (G)SIRT1 after 12 weeks of TXL treatment. Data are mean±SEM; n = 10 rats per group, ^#P<0.05 and ^$P>0.05 vs. WKY group, respectively; ^*P<0.05 and ^&P>0.05 vs. SHR group, respectively. ERK, extracellular signal-regulated kinase; PI3K, phosphatidylinositol 3-kinase;AMPK, Adenosine monophosphate-activated protein kinase; WKY, Wistar-Kyoto.</p

Effects of Tongxinluo(TXL) on fibrotic mediators in spontaneously hypertensive rat(SHR) kidneys.

<p>Representative images and quantitative analysis of(A) α-SMA,(B)collagen IV and (C) fibronectin immunostaining. PCR analysis of(D)TGFβ1 and (E) SMAD3.(F)Quantitative analysis of tubulointerstitial fibrosis. Scale bar, 20 μm. Data are mean±SEM; n = 10 rats per group, ^#P<0.05 and^*P<0.05 vs. WKY and SHR groups, respectively. α-SMA, α-smooth muscle actin; PCR, polymerase chain reaction; TGFβ1,transforming growth factor β1;SMAD3, small mothers against decapentaplegic homolog 3;WKY, Wistar-Kyoto.</p