DNA methyltransferase 3A regulates the expressions of collagens during the activation of cardiac fibroblasts / 医学研究生学报

Objective The mechanisms of methylation acting on myocardial fibrosis are not yet clear at present. The aim of this study was to investigate the role of DNA methyltransferase 3A (DNMT3A) in regulating the expressions of collagens during the activation of cardiac fibroblasts.Methods Cardiac fibroblasts were obtained from 50 neonatal mice and divided into three groups: blank control, DNMT3A overexpression plasmid (mDNMT3A-pEGFP-C3) and small interference DNMT3A siRNA. The contents of collagens in the cell supernatant were detected by ELISA. The mRNA and protein expressions of type I collagen (Col Ⅰ), type Ⅲ collagen (Col Ⅲ) and DNMT3A in the cardiac fibroblasts were determined by real-time quantitative PCR and Western blot respectively and the proliferative activity of the cardiac fibroblasts measured by CCK8 assay.Results The contents of Col I and Col Ⅲ in the cell supernatant were significantly increased in the DNMT3A overexpression plasmid group but decreased in the DNMT3A siRNA group as compared with those in the blank control (P<0.05). The expressions of Col Ⅰ, Col Ⅲ and DNMT3A were remarkably higher in the DNMT3A overexpression plasmid group but lower in the DNMT3A siRNA group than in the blank control (P<0.05). The cell activity was markedly higher in the DNMT3A overexpression plasmid group than in the empty vector plasmid and control groups (2.087±0.317 vs 1.063±0.223 and 1.082±0.207, P<0.05) but lower in the DNMT3A siRNA group (0.463±0.087) than in the latter two (P<0.05).Conclusion DNMT3A can increase the proliferation and activation of cardiac fibroblasts, upregulate the expressions of collagens and thus promote myocardial fibrosis.

Protective effect of IL-17 monoclonal antibody on rats with pulmonary fibrosis and its partial mechanisms / 中国药理学通报

Aim To explore the protective effects of in-terleukin-17 ( IL-17 ) monoclonal antibody ( mAb ) on bleomycin-induced pulmonary fibrosis rats and the re-lated mechanisms. Methods Seventy-five male SD rats were randomly divided into normal control group, sham operation group, model group, non-specific IgG group and IL-17 mAb group. Each group included fif-teen rats. Rats in the latter three groups were intratra-cheally administered with bleomycin A5 to establish pulmonary fibrosis model, whereas the ones in sham operation group were treated with the same volume of physiological saline. On day 7, 14 and 21, rats in non-specific IgG group and IL-17 mAb group were in-jected with non-specific IgG and IL-17 mAb, respec-tively,through the caudal vein. However,the ones in the other groups were administered with the same volume of physiological saline. All rats were sacrificed on day 28. Pulmonary tissues were then removed, and HE and Masson staining was performed. The contents of IL-17, IL-6 and tumor necrosis factor-α ( TNF-α) in pulmo-nary tissues were measured by enzyme linked immu-nosorbent assay ( ELISA ) . Western blot was used to analyze the pulmonary tissues protein expression of nu-clear factor-κB ( NF-κB) p65 in the nucleus as well as collagen type I ( Col Ⅰ) and collagen type III ( ColⅢ) in the whole cells. The levels of Col Ⅰ and ColⅢ in the pulmonary tissues were detected by fluores-cence real-time quantitative PCR. Serum was separa-ted, and the concentrations of procollagen type 1 carboxyterminal propeptide ( PICP ) and procollagen type III aminoterminal propeptide ( PIIINP ) in serum were then measured by ELISA. Results The severity of alveolitis and pulmonary fibrosis was lower in IL-17 mAb group than that in model group and non-specific IgG group ( P 0. 05 ) . Similar results were also seen between sham operation group and normal control group ( P >0. 05 ) . Conclusion IL-17 mAb protects rats from pulmonary fibrosis by inhibiting inflammatory response via downregulating NF-κB expression and decreasing collagen synthesis in the pulmonary tissues.

Effect of Tangnaikang on Extracellular Matrix Components of Human Renal Tubular Epithelial Cell HK-2 Induced by TGF-β1 / 中国中医药信息杂志

Objective To explore the effect of Tangnaikang (TNK) on extracellular matrix expression of human tubular epithelial cell HK-2 induced by TGF-β1 and explore its mechanism on the renal fibrosis. Methods The HK-2 cells were cultured by DMEM/F12 (1∶1) with 10%fetal bovine serum and divided into control group, TGF-β1 group (TGF-β110 ng/mL), rat serum control group (TGF-β110 ng/mL +10% rat serum), TNK-containing rat serum therapy groups (TGF-β110 ng/mL+5% Tangnaikang, or +10%Tangnaikang, or +20% TNK). After 24 h of administration, the expression of ColⅠ, Col Ⅲ and FN mRNA were tested by fluorescence quantitative PCR assay. Results The expression of ColⅠ, Col Ⅲ and FN mRNA of HK-2 cultured with TGF-β1 were much higher than the control, and significantly decreased in HK-2 cultured with TGF-β1 plus Tangnaikang compared with only TGF-β1 (P<0.05), but rat serum control had no effect. Conclusion TNK could inhibit the expression of ColⅠ, Col Ⅲ and FN mRNA of HK-2 cell induced by TGF-β1, and prevent the development of renal fibrosis to some extent.

Effect of Moldavica Total Flavone on Pulmonary Function in Ovalbumin-induced Bronchial Asthmatic Rats / 中国中医药信息杂志

Objective To investigate the effect of moldavica total flavone on pulmonary function and collagen type Ⅲ in lung tissue of ovalbumin-induced bronchial asthmatic rats. Methods A total of 60 SD rats were randomly divided into control group, model group, dexamethasone group, and moldavica total flavone low, medium and high dose group. Rat asthma model was established by ovalbumin challenge methods except the control group. The control and model group were intragastrically given distilled water, medicine groups were intragastrically given moldavica total flavone and dexamethasone for 30 d, once per day. After the last administration, the pulmonary function and airway responsiveness including breathing frequency (F), minute ventilation (MVb), peak inspiratory flow (PIFb), peak expiratory flow (PEFb) and enhanced pause (Penh) were measured by using non-invasive measurement system (Penh system, Buxco, USA). The collagen type Ⅲ in lung tissue were detected by ELISA. Results The moldavica total flavone at 180 and 360 mg/kg decreased F and Penh (P<0.05, P<0.01), increased MVb, PIFb and PEFb (P<0.05), and decreased collagen type Ⅲ in lung tissue (P<0.05, P<0.01). Conclusion Moldavica total flavone can relieve the airway hyperresponsiveness and remodeling, improve respiratory function of ovalbumin-induced bronchial asthmatic rats.

Influence of Tongfu Xiezhuo Decoction on the expression of ?-SMA,LN,ColⅢ in tubulo interstitial fibrosis rats / 中成药

AIM:To investigate the possible preventive mechanism of Tongfu Xiezhuo Decoction(Radix astragali,Radix et Rhizoma rhei,Rhizoma acori tatarinowii,Semen pharbitidis,Radix aconiti lateralis praeparata,Radix et Rhizoma ginseng, Rhizoma atractrylodis macrocephale,Radix et Rhizoma Salviae mltiorrhizae,Semen Vaccariae,Pericarpium citri reticulate) for the tubule interstitial fibrosis by influence on the expression of ?-SMA,LN,ColⅢ in rats. METHODS: Unilateral ureteral occlusion(UUO) rats were randomly divided into 6 groups,including Sham,UUO,Enalapril group and Tongfu Xiezhuo Decoction of high-dosage,mid-dosage and low-dosage groups,the same amount of physiological saline for intragastric administration was administered by gavage.Sera and the kidney tissues were collected from each group on the twelfth day,Scr and BUN were measured.HE,VG staining measurement of tubulo-interstitial damage index and collagen deposition level,immuneohistochemical studies localizing ?-smooth muscle,actin(?-SMA),laminin-(LN),collagenⅢ(ColⅢ)were carried out. RESULTS: In UUO rats,the tubular interstitial damage index,the expressions of ?-SMA,laminin-(LN),collagenⅢ(ColⅢ) were all increased compared with those of Sham group(P

Effect of all-trans-retinoic acid on tubulointerstitial macrophage infiltration in rats with renal tubulointerstitial fibrosis / 西安交通大学学报(医学版)

Objective To investigate the effect of at-RA in macrophage accumulation in tubulointerstitium of rats with renal tubulointerstitial fibrosis.Methods Unilateral ureteral obstructive(UUO) rat animal models were used for the study.40 SD rats were randomly divided into 5 groups: sham group,UUO group,benazepril group,low-dose at-RA groups and high-dose at-RA groups.The rats were under intragastric administration by benazepril(10mg/(kg?d)) in benazepril group,and by(at-RA)(10mg/(kg?d)) in low-dose at-RA group and 20mg/(kg?d) in high-dose at-RA group and by sodium chloride in tales doses in sham group and UUO group from the day before the operation to 14 day after operation.Immunohistochemistry staining of CD68 and Col Ⅲ was used to define the macrophage accumulation and expression of interstitial Col Ⅲ.The degree of tubulointerstitial damage was scored by HE and Masson staining.Results Tubulointerstitial macrophage infiltration were all significantly reduced by(at-RA) or benazepril treatment.They also improved the histological changes of UUO rats and inhibited interstitial colⅢ deposition.Conclusion Reduction of interstitial macrophage infiltration may be an important event by which(at-RA) or benazepril prevents renal injury caused by UUO.