Yang Cao — Page 5 | PubMed Health

PubMed2002

Effect of transforming growth factor-beta(2) on extracellular matrix synthesis in bovine trabecular meshwork cells.

OBJECTIVE: To investigate the effect of transforming growth factor-beta(2) (TGF-beta(2)) in human aqueous humor on extracellular matrix synthesis in cultured bovine trabecular meshwork cells. METHODS: Cultured 3-5 passage bovine trabecular meshwork cells were divided into control group and experimental group. After treated with 0 ng/L (control), 0.32 x 10(3) ng/L, 1.00 x 10(3) ng/L, 3.20 x 10(3) ng/L TGF-beta(2) for 48 h, collagen, fibronectin and hyaluronic acid synthesis in bovine trabecular meshwork cells were examined respectively by (3)H-proline incorportion and liquid scintillation technique, enzyme linked immunosorbent assay (ELISA) and radioimmunoassay (RIA). RESULTS: In comparison with the control group, TGF-beta(2) significantly promoted the synthesis of collagen at 0.32 x 10(3) ng/L (P < 0.05), 1.00 x 10(3) ng/L (P < 0.01), 3.20 x 10(3) ng/L (P < 0.01), (3)H-proline incorporation increased in a concentration-dependent manner; and 1.00 x 10(3) ng/L (P < 0.05), 3.20 x 10(3) ng/L (P < 0.01) TGF-beta(2) significantly promoted the synthesis of fibronectin in cultured bovine trabecular meshwork cells. 1.00 x 10(3) ng/L (P < 0.01) and 3.20 x 10(3) ng/L (P < 0.01) TGF-beta(2) significantly inhibited the synthesis of hyaluronic acid in the cultured cells. CONCLUSION: The data suggest that TGF-beta(2) play important roles in extracellular matrix age-related changes of normal trabecular meshwork and abnormal deposition of the extracellular matrix in the trabecular meshwork, especially the juxtacanalicular tissue of primary open-angle glaucoma patients.

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PubMed2002

[Studies of the clone and cell expression of human lactoferritin gene].

In this paper,we directly cloned 2.366 Kb cDNA sequence of human lactoferrin gene and 800 bp 5'flank regulatory sequence of beta/lactoglobulin gene from goat by PCR,then connected them with the expression vector pLNCX. The recombinant plasmid pLNCXHLF containing human lactoferrin gene cDNA was transfected into mice mammary tumor cell line MA/782 after liposome transinfection. Positive single clone cells were selected with G418 and by PCR. After proliferating,the transfected cells immobilized and cultured in sodium alginate were induced by hormone. The result of Western blotting analysis on cultured cell supernatant shows that transfected cells can express the exogenic gene and secrete hLF protein, whose MW is 34 KD. The highest amount detected by ELISA reached 65 mg/l medium/10(5) cells. The result of antibacterial experiment indicates that the recombinant hLF protein has the effect of inhibiting E.coli proliferation;moreover,its activity is superior to the commercial available hLF's.

English Abstract↗