ABSTRACT
This study was aimed to observe the effects of Gui Curcuma extract on proliferation, collagen production, and matrix metalloproteinase (MMP) expression by hepatic stellate cells (HSC-LX2). HSC-LX2 were cultured in DMEM broth (10% FBS), then treated with Gui Curcuma extract (with the concentration of 0, 22.5, 45, 90 μmol·L-1) for 24 h. The proliferation of HSC-LX2 was measured by MTT assay; injury in HSC-LX2 was indicated with LDH re-lease; collagen I production was detected by ELISA; and MMP-l expression was determined by Western blot. The re-sults showed that Gui Curcuma extract (with the concentration of 22.5, 45, 90 μmol·L-1) dose-dependently inhibited proliferation of HSC-LX2 by 16.2%, 43.9%, 59.4% in MTT assay, respectively. The IC50 was 59.1 μmol·L-1. There was no significant impact on LDH release. The Gui Curcuma extract (with the concentration of 45, 90 μmol·L-1) can significantly reduce the expression of collagen I level (P< 0.01), whereas MMP-1 expression was significantly uplift-ed (P< 0.05) by Gui Curcuma extract (with the concentration of 45, 90 μmol·L-1) in Western blot. It was concluded that Gui Curcuma extract had the potential to prevent hepatic fibrosis, through inhibiting proliferation and collagen I production, and increasing MMP-1 protein expression.
ABSTRACT
Objective According to the distribution of low selenium areas, low nutrition state of the residents and the affecting cartilage growth and articular cartilage of Kashin-Beck Disease(KBD),the chondrocyte differentia- tion and differential expression of collagen types Ⅰ , Ⅱ and Ⅹ in articular cartilage from Chinese mini-pigs treated with low selenium were investigated in order to gain insight into the effects of these conditions on chondrocyte differ- entiation in KBD cartilage. Mothods Eleven male juvenile mini-pigs, aged from 4 weeks to 6 weeks after birth, were divided into 3 groups. The Se content in the diet of the “low Se” group was 0. 035mg/kg diet, and 0. 175 mg/kg diet in the control. For Se-supplemented group 0. 390mg /kg diet was added. The content of Se in blood was assayed at the beginning and at the end of each experiment. Samples of articular cartilage were taken from the right femur condylus, and collagen types Ⅰ , Ⅱ and Ⅹ in articular cartilage were analyzed by immunohistochemistry and in situ hybridization. Results①All cartilage samples from juvenile mini-pigs fed with low selenium diet revealed a re- duction in type Ⅹ collagen mRNA expression in the hypertrophic chondrocytes as shown by in situ hybridization, and reduced type Ⅹ collagen deposition in the lower hypertrophic zone as shown by immunohistochemistry. ②Addition of selenium to the diet restored the type Ⅹ collagen to normal level. ③Type Ⅱ collagen was evenly distributed over the entire articular cartilage in all experimental and control groups. Type Ⅱ collagen mRNA signals were most prominent in the upper articular layer as well as in the hypertrophic zone in all groups. Type Ⅱ collagen expression was restrict- ed to the zone of endochondral ossification in all experimental groups and the control. Conclusion Low selenium has an down-regulatory role on the synthesis and deposition of collagen type Ⅹ in hypertrophic chondrocytes in articular cartilage of mini-pigs. Supplement of the low Se diet with additional Se restored the signals of collagen type Ⅹ to nor- mal levels. These findings indicate that selenium deficiency may disturb chondrocyte differentiation to hypertrophic cells in the growth plate,and worthy to be investigated further.