Antifibrotic effects of crocetin in scleroderma fibroblasts and in bleomycin-induced sclerotic mice

OBJECTIVE: To investigate the antifibrotic effects of crocetin in scleroderma fibroblasts and in sclerotic mice. METHODS: Skin fibroblasts that were isolated from three systemic scleroderma (SSc) patients and three healthy subjects were treated with crocetin (0.1, 1 or 10 μM). Cell proliferation was measured with an MTT assay. Alpha-smooth muscle actin was detected via an immunohistochemical method. Alpha 1 (I) procollagen (COL1A1), alpha 1 (III) procollagen (COL3A1), matrix metalloproteinase (MMP)-1 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 mRNA levels were measured using real-time PCR. SSc mice were established by the subcutaneous injection of bleomycin. Crocetin (50 mg/kg/d) was injected intraperitoneally for 14 days. Dermal thickness and lung fibrosis were assessed with Masson's trichrome staining. Plasma ET-1 was detected with an enzyme-linked immunosorbent assay (ELISA). Skin and lung ET-1 and COL1A1 mRNA levels were measured via real-time PCR. RESULTS: Crocetin inhibited the proliferation of SSc and normal fibroblasts, an effect that increased with crocetin concentration and incubation time. Crocetin decreased the expression of α-SMA and the levels of mRNA for COL1A1, COL3A1 and matrix metalloproteinase-1, while crocetin increased TIMP-1 mRNA levels in both SSc and normal fibroblasts. Skin and lung fibrosis was induced, and the levels of ET-1 in the plasma, skin and lungs were elevated in bleomycin-injected mice. Crocetin alleviated the thickening of the dermis and lung fibrosis; decreased COL1A1 mRNA levels in the skin and lung; and simultaneously decreased ET-1 concentrations in the plasma and ET-1 mRNA levels in the skin and lungs of the bleomycin-induced sclerotic mice, especially during the early phase (weeks 1-3). CONCLUSION: Crocetin inhibits cell proliferation, differentiation and collagen production in SSc fibroblasts. Crocetin alleviates skin and lung fibrosis in a bleomycin-induced SSc ...

Relationship between serum concetrations of type III procollagen, hyluronic acid and histopathological findings in the liver of HCV-positive blood donors

BACKGROUND: Serologic markers have been proposed for monitoring hepatic fibrosis in chronic liver disease. Among fibrosis markers, type III procollagen (PIIIP) and hyaluronic acid have been studied in these patients. AIM: To evaluate the association between these serum markers with histological findings. METHODS: A prospective cross-sectional study was carried out with HCV-positive blood donors. The studied population included men and women whose age ranged from 18 to 60 years, with elevated liver function tests [ALT levels > 1.5 times the normal value and alterations of two or more of the following: any changes in the levels of ALT, aspartate aminotransferase, conjugated bilirrubin, gammaglobulin, gammaglutamyltranspeptidase, albumin, platelet count; alkaline phosphatase levels >1.5 times the normal value, or prothrombin time below 70 percent and above 60 percent]. Fourty-nine patients were submitted to liver biopsy, blood analysis of PIIIP, hyaluronic acid, besides liver function tests. RESULTS: Liver function tests were not associated with tissular fibrosis, as assessed by ALT (>1.5 times above normal, fibrosis risk=18.8 percent; <1.5 times, 11.8 percent). Elevated PIIIP was correlated with 66.7 percent chance of fibrosis, whereas normal levels, 9.3 percent. Hyaluronic acid, when elevated, gave a chance of 33.3 percent of fibrosis; when normal, 12.5 percent. CONCLUSION: There was no association between liver function tests, hyaluronic acid and fibrosis. However, PIIIP was related with liver fibrosis. Maybe, this marker should be useful to assess fibrosis in patients with chronic hepatitis C.

RACIONAL: Marcadores sorológicos têm sido propostos para monitorar fibrose hepática em doença crônica do fígado. Dentre os marcadores de fibrose, ácido hialurônico e procolágeno tipo III têm sido estudados nestes pacientes. OBJETIVO: Avaliar a associação de marcadores séricos de fibrose com achados histológicos. MÉTODOS: Foi realizado estudo transversal prospectivo em doadores de sangue anti-HCV positivos. A população estudada incluiu homens e mulheres com idade entre 18-60 anos com provas de função hepática alteradas (níveis de alanina aminotransferase >1.5 vezes do normal e alterações de dois ou mais dos seguintes: qualquer alteração nos níveis de alanina aminotransferase, aspartato aminotransferase, bilirrubina conjugada, gamaglobulina, gamaglutamiltranspeptidase, albumina, plaquetas, níveis de fosfatase alcalina >1,5 vezes o valor normal, tempo de protrombina abaixo de 70 por cento e acima de 60 por cento). Quarenta e nove pacientes foram submetidos a biopsia hepática e coleta de sangue para análise de procolágeno tipo III, ácido hialurônico e provas funcionais hepáticas. RESULTADOS: Não houve relação entre elevação de provas de função hepática e a presença de fibrose - ALT (>1,5 vezes acima do normal, risco de fibrose = 18,8 por cento; <1,5 vezes, 11,8 por cento). Procolágeno tipo III elevado foi correlacionado com 66,7 por cento chances de fibrose, enquanto nível normal, 9,3 por cento. Ácido hialurônico, quando elevado, demonstrou chance de 33,3 por cento de fibrose; quando normal, 12.5 por cento. CONCLUSÕES: Não houve associação entre provas de função hepática, ácido hialurônico e fibrose, mas houve entre esta última e procolágeno tipo III. Talvez este marcador possa ser útil para avaliar fibrose em pacientes com hepatite crônica pelo vírus C.

Assessment of liver dysfunction in silica exposed workers

Seventy five silica exposed workers in sandbricks industry and twenty five controls were chosen for this study. Each individual was subjected to a questionnaire [for personal and medical histories] and clinical examination. Liver function tests, procollagen III peptide, alpha-I - antitrypsin, hepatitis markers [B and C], haematological tests for schistosomiasis, urine and stool analysis, abdominal sonography and chest X-ray were done for all chosen subjects. Environmental study was done for estimation of total and free crystalline silica. The results of environmental study showed higher concentration of total respirable dust and free crystalline silica than threshold limit values. Silica exposed workers had a significantly higher mean of gamma-glutamyl trans peptidase and procollagen III peptide than controls. These two parameters increased significantly in exposed workers [free from schistosomiasis and / or hepatitis markers] as chest grading of silicosis and time intensity factor increased. Gamma-glutamyl-trans peptidase and procollagen III peptide can be used for early detection of liver dysfunction in silica exposed workers. Also, in the pre-employment medical examination, individuals with previous liver affection [either due to schistosomiasis and / or hepatitis] should not be exposed to silica to avoid further deterioration of liver function