VPA alleviates bleomycin-induced pulmonary fibrosis in rats

AIM: To investigate the effect of sodium valproate (VPA) on bleomycin-induced pulmonary fibrosis (PF) and its mechanism. METHODS: SD rats (n=42) were randomly divided into model group and treatment group. Bleomycin at dose of 5 mg/kg was intratracheally injected to establish a rat PF model. The rats in treatment group were given normal saline (NS, 0.5 mL/d), VPA (300 mg·kg^-1·d^-1) or dexamethasone (DEX, 0.6 mg·kg^-1·d^-1) via intraperitoneal injection from the 14th day to the 28th day after modeling. The rats in model group were sacrificed on 0 d, 14 day and 28 d after modeling . The rats in treatment group were killed at 14th day after treatment. The effects of VPA on PF were evaluated by HE and Masson staining. The hydroxyproline (HYP) content in the rat lung tissues was detected, and the expression of α-smooth muscle actin (α-SMA) and E-cadherin was determined by Western blotting. RESULTS: HE staining showed that the alveolar structure and interstitial morphology in VPA group were better than those in NS group and DEX group. The level of collagen in VPA group was significantly lower than that in DEX group and NS group by determining the HYP content and Masson staining. VPA reduced the expression of α-SMA, a mesenchymal marker protein of PF, while increased the expression of epithelial marker protein E-cadherin. CONCLUSION: VPA reduces collagen content and distribution, and up-regulates the expression of the epithelial marker protein E-cadherin, while down-regulates mesenchymal marker protein α-SMA, thereby preventing the rat lung tissues from bleomycin-induced PF.     

Role of mineralocorticoid receptor in bleomycin-induced pulmonary fibrosis

AIM:To investigate the role of mineralocorticoid receptor (MR) in the lungs of experimental fibrotic mice. METHODS:C57BL/6 male mice (6～8 weeks old) were randomly divided into control group, bleomycin treatment group (Bleo) and bleomycin+spironolactone treatment group (Bleo+Spiro). For induction of pulmonary fibrosis, the mice were administered bleomycin at dose of 2.5 mg/kg dissolved in 50 μL saline by the intratracheal route or given 50 μL sterile saline as control. The mice in Bleo+Spiro group were treated with spironolactone (20 mg/kg) daily by oral gavage throughout the experiment. The mice were sacrificed at 12 h, 1 d, 2 d, 3 d, 7 d, 14 d and 28 d after administration of bleomycin. HE staining and Masson’s trichrome staining were used to conduct histopathologic examination. The mRNA expression levels of collagen 1 (Col1), collagen 3 (Col3), transforming growth factor beta (TGF-β), monocyte chemoattractant protein 1 (MCP-1), and MR were examined by real-time PCR. RESULTS:The results of histological analysis revealed the classical pathological stages of bleomycin-induced lung fibrosis, including acute inflammation phase (from 12 h to 3 d), progressive fibrosis phase (14 d) and late fibrosis phase (28 d). Compared with Bleo group, the inflammatory responses of the lungs in Bleo+Spiro group were attenuated in the acute inflammation phase and the degree of fibrosis was significantly reduced at 14 d after administration of bleomycin. Treatment with spironolactone effectively down-regulated the mRNA expression of MR. The levels of MCP-1 (in the acute inflammation phase), TGF-β (at 14 d), Col1 and Col3 (at 14 d) were also significantly reduced. CONCLUSION: Blockage of MR significantly attenuates the degree of bleomycin-induced pulmonary fibrosis by regulating the production and secretion of MCP-1 and TGF-β, thus reducing the degree of inflammation and inhibiting the expression of TGF-β in the progressive fibrotic phase.     

Electrochemotherapy in treatment of canine oral non‐tonsillar squamous cell carcinoma. A case series report

Non‐tonsillar squamous cell carcinoma (ntSCC) is a common and locally aggressive oral tumour in dogs. The treatments of choice are currently surgery and radiotherapy. Electrochemotherapy (ECT) is a local ablative anti‐tumour technique using electric pulses to enhance the intracellular diffusion of cytotoxic drugs. The aim was to retrospectively evaluate the outcome of patients with oral ntSCC treated with ECT. Twelve dogs with ntSCC were retrospectively enrolled. ECT was combined with IV bleomycin (15 000 UI/m²) alone in 11 cases and post‐surgery in 1. Parameters considered were: tumour site and size, electroporation parameters, response rate (complete remission [CR], partial remission [PR]), median survival time (MST), recurrence rate (RR), median disease‐free interval (DFI) and treatment toxicity (6‐point scale). Median tumour size was 1.65 cm (range 0.3‐8.0 cm) and the response rate was 90.9% (10/11; 8 CR and 2 PR). Two dogs underwent a second ECT. MST for dogs dead with tumour (n = 2) was 110 days and for dogs dead without tumour (n = 3) was 831 days. Among five surviving dogs, one experienced tumour recurrence and four were in CR. Results from two dogs were analysed separately. Overall RR was 27.3%. DFI and MST for dogs with recurrence were 50 and 115 days, respectively. Treatment toxicity was very low. We noticed that all dogs with tumours smaller than 1‐2 cm achieved CR without recurrence suggesting a favourable prognosis when using ECT. ECT for canine ntSCC could be considered a valid treatment option especially for smaller tumours, but a larger caseload would be needed to confirm this statement.     

低剂量与高剂量顺铂联合5-氟脲嘧啶及平阳霉素治疗晚期鼻咽癌的疗效比较

目的：比较低剂量与高剂量顺铂（ＤＤＰ）联合５－氟脲嘧啶（５－Ｆｕ）、平阳霉素（ＢＬＭ）（简称ＰＦＢ方案）对晚期鼻咽癌的疗效及毒性反应。方法：经病理学确诊的放疗后复发或有远处转移的晚期鼻咽癌３８例,随机分为单次高剂量ＤＤＰ组（ＨＤ－ＤＤＰ组）１７例与低剂量ＤＤＰ连续５ｄ静滴组（ＬＤ－ＤＤＰ组）２１例。ＨＤ－ＤＤＰ组予ＤＤＰ１００ｍｇ／ｍ２,第１天静脉输入,配合水化;ＬＤ－ＤＤＰ组予ＤＤＰ２０～３０ｍｇ／ｍ２第１～５天静脉输入,不予水化。两组同时联合５－Ｆｕ、ＢＬＭ,每３～４周重复。结果：总有效率（完全缓解＋部分缓解）：ＨＤ－ＤＤＰ组５２．９％,ＬＤ－ＤＤＰ组４７．６％（Ｐ＞０．０５）。对不同转移部位有效率以肺转移疗效较好（５８．３％、６０％）,肝转移无效。毒性反应：Ⅱ度以上呕吐发生率ＨＤ－ＤＤＰ组９０．４％,明显高于ＬＤ－ＤＤＰ组的４８．１％（Ｐ＜０．００１）;ＨＤ－ＤＤＰ组与ＬＤ－ＤＤＰ组的白细胞降低发生率分别为５２．３％、３３．３％（Ｐ＞０．０５）;肾毒性发生率分别为７．１％、５．５％（Ｐ＞０．０５）。结论：低剂量ＤＤＰ连续５ｄ静滴的ＰＦＢ方案疗效与高剂量ＤＤＰ的ＰＦＢ方案疗效接近,毒性反应轻微,应用简便     

Matrine attenuates bleomycin-induced pulmonary injury partially via modulating mononuclear phagocyte phenotype switching in mice

AIM: To investigate the influence of matrine (MA) on the phenotype switching of mouse monocytes and alveolar macrophages induced by bleomycin (BLM).METHODS: All mice were randomly divided into normal saline (NS) group, BLM group, BLM+NS group and BLM+MA group. The mice were administered with BLM at 2.5 mg/kg via oropharyngeal instillation. The mice in BLM+MA group were treated with MA (15 mg·kg^-1·d^-1) by oral gavage following BLM administration. The mice were sacrificed on days 3, 7, 14, and 21. The lungs were removed for pathological analysis. The circulating monocyte subsets and polarization state of bronchoalveolar lavage fluid (BALF)-derived alveolar macrophages were analyzed by flow cytometry.RESULTS: The results of HE and Masson trichrome staining in BLM and BLM+NS groups exhibited classical pathological stages of lung fibrosis, including acute inflammation phase and later fibrosis phase. Compared with BLM+NS group, MA treatment alleviated the inflammatory response and the degree of fibrosis induced by BLM (P<0.05). There was a rapid change of circulating Ly6C^hi monocytes and its magnitude was positively associated with the pulmonary inflammatory response. An expansion of M2-like alveolar macrophages was positively correlated with the magnitude of lung fibrosis. Moreover, MA treatment partially normalized the phenotype switching of monocytes and alveolar macrophages.CONCLUSION: Matrine treatment attenuates BLM-induced pulmonary injury partially via modulating the phenotype switching of monocytes and alveolar mocrophages.     

Caffeic acid phenethyl ester and vitamin E moderates IL-1β and IL-6 in bleomycin-induced pulmonary fibrosis in rats

The aim of this study was to investigate the effects of Caffeic acid phenethyl ester (CAPE), which has been demonstrated to have antiinflammatory, antiproliferative, anticancerogenic, and antioxidant effects, and vitamin E on IL-1β and IL-6 in bleomycin-induced (BLM-induced) pulmonary fibrosis in rats. Thirty-two Sprague-Dawley rats were divided randomly into four groups as untreated control, bleomycin, bleomycin + CAPE, and bleomycin + vitamin E groups. At the end of the treatment, blood IL-1β and IL-6 levels were quantified. Bleomycin application to the rats resulted in a significant increase in the cytokine levels as compared to the controls. Administration of CAPE and vitamin E prevented the increase of blood IL-1β and IL-6 levels compared to the rats treated with bleomycin alone. Data presented here suggest that CAPE and vitamin E play a protective and moderator role against BLM-induced lung injuries by decreasing the primary inflammatory cytokines, such as IL-1β and IL-6.     

Etanercept attenuates bleomycin-induced lung fibrosis in mice

AIM: To evaluate the effect of tumor necrosis factor α (TNF-α) antagonist etanercept on bleomycin-induced lung fibrosis in mice.METHODS: Forty-five Kunming female mice were randomly divided into 3 groups: the mice in control group were intraperitoneally injected with vehicle and intratracheally administered with saline aerosol, the mice in bleomycin group were intraperitoneally injected with vehicle and intratracheally administered with bleomycin (3 mg/kg) aerosol, and the mice in bleomycin+etanercept group were intraperitoneally injected with etanercept (4 mg/kg) every 3 d and intratracheally administered with bleomycin aerosol. All animals were sacrificed 28 d after treatments. The left lung was fixed in 10% neutral formalin and then stained with hematoxylin-eosin or Masson’s trichrome for the pathological examination. The tissues of right lung were sampled for measuring the content of hydroxyproline (HYP) by the method of alkaline hydrolysis. The serum concentrations of TNF-α and TGF-β were detected by ELISA. Total tissue protein was extracted for examination of ERK1/2, JNK and p38 by Western blotting.RESULTS: Etanercept prevented the collagen accumulation under the airway epithelium and decreased the scores of lung inflammation and fibrosis induced by bleomycin with significantly reduced the levels of tissue HYP, serum TNF-α and serum TGF-β. The protein phosphorylations of ERK/JNK/p38 in the lung tissues were remarkably decreased compared with BLM group.CONCLUSION: Etanercept decreases the phosphorylations of ERK1/2/JNK/p38 via inhibiting the expression of TNF-α and TGF-β. Etanercept might be useful in the treatment of pulmonary fibrosis.     

Gefitinib inhibits epithelial-mesenchymal transition by up-regulating Foxo3a expression in mice with bleomycin-induced lung fibrosis

AIM：To identify the effect of gefitinib on the expression of forkhead box protein O3a (Foxo3a), α-smooth muscle actin (α-SMA) and related signal pathway molecules in the mice with bleomycin-induced lung fibrosis and to investigate the inhibition mechanism of gefitinib on lung epithelial-mesenchymal transition. METHODS：Thirty Kunming female mice were randomly divided into 3 groups:control group (received normal saline intratracheally), bleomycin group (received bleomycin intratracheally, 3 mg/kg), and bleomycin plus gefitinib group (received bleomycin intratracheally and gefitinib orally, 20 mg/kg). All the mice were sacrificed 14 d after the treatments. Pulmonary histological changes were evaluated by hematoxylin-eosin staining and Masson trichrome staining. The mRNA levels of Foxo3a and α-SMA in the lung tissues were detected by RT-PCR. Nuclear Foxo3a, α-SMA, and phosphorylation of EGFR, Akt and Foxo3a in the lung tissues were determined by Western blotting. RESULTS：Gefitinib inhibited bleomycin-induced lung fibrosis and significantly decreased the scores of lung inflammation and fibrosis. Foxo3a mRNA expression and total Foxo3a protein expression were increased, while the phosphorylated Foxo3a was decreased. Nuclear Foxo3a was increased significantly. Meanwhile, phosphorylated EGFR and Akt were decreased. The level of α-SMA was observably increased. CONCLUSION:Gefitinib restores Foxo3a activity and reduces α-SMA expression by modulating EGFR/Akt activity, thus inhibiting bleomycin-induced lung fibrosis.