OBJECTIVE: To investigate the influence of beta-Elemene on expression of ANG II and RhoA/ROCK signaling in Hepatic Stellate Cells. METHOD: In vitro, HSC-T6 cell line was cultured for 24 hours and treated with several concentration of beta-elemene (5.0, 5.0, 2.5 mg x L(-1)) and Y-27632 (30 micromol x L(-1)) for 4, 12 and 24 h. Secretion of ANG II in the supernatant was detected by Radioimmunoassay. The mRNA expression of AGT, RhoA, ROCK-1 and ROCK-2 for 4 h, 12 h, 24 h was detected by RT-PCR respectively. RESULT: On the time point of 4h, the secretion of ANG II in supernatant by 10 mg x L(-1) beta-elemene was 50.970 +/- 8.081 pmol x L(-1), vs the control group (74.500 +/- 10.999) pmol x L(-1), P < 0.05; 5.0 mg x L(-1) and 2.5 mg x L(-) beta-elemene had no inhibitory effect on the secretion of ANG II, P > 0.05. On the time point of 12h, the secretion of ANG II in supernatant by 10 mg x L(-1), 5 mg x L(-1) beta-elemene was 83.727 +/- 6.850 pmol x L(-1), 91.090 +/- 3.226 pmol x L(-1), respectively, lower than the control (104.367 +/- 5.030 pmol x L(-1)), P < 0.01, P < 0.05. On the time point of 24h, compared with the control (116.620 +/- 7.110) pmol x L(-1)), the secretion ofANGII in supernatant by 2.5 mg x L(-1) and 5 mg x L(-1) beta-elemene was (104.133 +/- 3.296) pmol x L(-1), (100.957 +/- 2.581) pmol x L(-1), respectively, P < 0.05, P < 0.01; but the effect of 10 mg x L(-1) beta-elemene was not obviouse, P > 0.05. Compared with control group, the mRNA expression of AGT by different concentration of beta-elemene were significantly inhibited on different time point (4, 12, 24 h), F value was 30.33, 28.04, 107.19, respectively, P = 0.000. On the time point of 4h, 12 h and 24 h, the RhoAmRNA expression could be inhibited by 2.5, 5.0, 10 mg x L(-1) beta-elemene, (P = 0.000), and there existed no dose dependence. On the time point of 4 h and 24 h, ROCK-1 and ROCK-2mRNA could be inhibited by 2.5, 5.0 and 10 mg x L(-1) beta-elemene, P < 0.01, but on the time point of 12 h, there was no inhibitory effect. CONCLUSION: Beta-elemene can inhibit the expression of AGTmRNA in HSC and the secretion of ANGII in supernatant. In addition, beta-elemene can inhibit the mRNA expression of RhoA, ROCK-1, ROCK-2mRNA to further regulate the biological effect of ANG II and delay the progress of hepatic fibrosis.
OBJECTIVE: To investigate the influence of beta-Elemene on expression of ANG II and RhoA/ROCK signaling in Hepatic Stellate Cells. METHOD: In vitro, HSC-T6 cell line was cultured for 24 hours and treated with several concentration of beta-elemene (5.0, 5.0, 2.5 mg x L(-1)) and Y-27632 (30 micromol x L(-1)) for 4, 12 and 24 h. Secretion of ANG II in the supernatant was detected by Radioimmunoassay. The mRNA expression of AGT, RhoA, ROCK-1 and ROCK-2 for 4 h, 12 h, 24 h was detected by RT-PCR respectively. RESULT: On the time point of 4h, the secretion of ANG II in supernatant by 10 mg x L(-1) beta-elemene was 50.970 +/- 8.081 pmol x L(-1), vs the control group (74.500 +/- 10.999) pmol x L(-1), P < 0.05; 5.0 mg x L(-1) and 2.5 mg x L(-) beta-elemene had no inhibitory effect on the secretion of ANG II, P > 0.05. On the time point of 12h, the secretion of ANG II in supernatant by 10 mg x L(-1), 5 mg x L(-1) beta-elemene was 83.727 +/- 6.850 pmol x L(-1), 91.090 +/- 3.226 pmol x L(-1), respectively, lower than the control (104.367 +/- 5.030 pmol x L(-1)), P < 0.01, P < 0.05. On the time point of 24h, compared with the control (116.620 +/- 7.110) pmol x L(-1)), the secretion ofANGII in supernatant by 2.5 mg x L(-1) and 5 mg x L(-1) beta-elemene was (104.133 +/- 3.296) pmol x L(-1), (100.957 +/- 2.581) pmol x L(-1), respectively, P < 0.05, P < 0.01; but the effect of 10 mg x L(-1) beta-elemene was not obviouse, P > 0.05. Compared with control group, the mRNA expression of AGT by different concentration of beta-elemene were significantly inhibited on different time point (4, 12, 24 h), F value was 30.33, 28.04, 107.19, respectively, P = 0.000. On the time point of 4h, 12 h and 24 h, the RhoAmRNA expression could be inhibited by 2.5, 5.0, 10 mg x L(-1) beta-elemene, (P = 0.000), and there existed no dose dependence. On the time point of 4 h and 24 h, ROCK-1 and ROCK-2mRNA could be inhibited by 2.5, 5.0 and 10 mg x L(-1) beta-elemene, P < 0.01, but on the time point of 12 h, there was no inhibitory effect. CONCLUSION:Beta-elemene can inhibit the expression of AGTmRNA in HSC and the secretion of ANGII in supernatant. In addition, beta-elemene can inhibit the mRNA expression of RhoA, ROCK-1, ROCK-2mRNA to further regulate the biological effect of ANG II and delay the progress of hepatic fibrosis.