Effects of ferritin heavy chain of Clonorchis sinensis on human hepatic stellate cell line LX-2 / 中华地方病学杂志

ABSTRACT

Objective To observe the effects of recombinant protein ferritin heavy chain of Clonorchis sinensis (CsFHC) on human hepatic stellate cell line LX-2. Methods LX-2 cells were cultured in vitro by the cell culture method . Cell proliferation/toxicity detection kit (CCK-8) was used to detect the cell proliferation activity of LX-2 cells stimulated with CsFHC recombinant protein [0 (control), 1, 2, 5, 10, 20 nmol/L] at 48 h and 72 h, respectively, and the effect of CsFHC recombinant protein on cell cycle was analyzed by flow cytometry; a semi quantitative RT-PCR was used to detect the mRNA expression of type I collagen (Collagen Ⅰ), type Ⅲ collagen (Collagen Ⅲ) and α smooth muscle actin (Αsma). Results The effects of CsFHC on proliferation of LX-2 cells (48 h 0.987 ± 0.042, 1.315 ± 0.105, 1.298 ± 0.078, 1.432 ± 0.089, 1.781 ± 0.040, 1.581 ± 0.056; 72 h1.050 ± 0.030, 1.503 ± 0.111, 1.671 ± 0.102, 1.769 ± 0.123, 1.927 ± 0.067, 1.492 ± 0.081) between groups were significantly different statistically (F = 1892.133, 534.136, P<0.05). The quiescent stage/DNA synthesis (G0/G1 ) cells in 1, 2, 5, 10, and 20 nmol/L CsFHC recombinant protein groups (74.93 ± 4.05, 75.87 ± 4.16, 76.73 ± 5.03, 78.57 ± 5.51, 74.90 ± 3.61) were significantly higher than that of control group (54.90 ± 3.61, P<0.05);the synthesis phase (S) + DNA synthesis late/mitotic (G2/M) cells (22.24 ± 3.06, 24.13 ± 2.00, 18.54 ± 1.53, 18.71 ± 1.53 and 21.17 ± 3.06) were lower than that of control group (33.26 ± 2.65,P < 0.05). In control and 1, 2, 5, 10, 20 nmol/L CsFHC recombinant protein groups, the differences of Mrna expression levels of Collagen Ⅰ, Collagen Ⅲ and α SMA were statistically significant (48 hF=81.419, 14.417, 70.456;72 hF=79.224, 50.461, 41.872, P<0.05). Conclusion The CsFHC recombinant protein can stimulate the proliferation and activation of LX-2 cells in vitro, the CsFHC involved in the process of hepatic fibrosis induced by Clonorchis sinensis.