Jin Zheng1,2, Li-Tian Ma2, Qin-You Ren2, Yue Hu3, Yang Bai4, Huan Bian5, Yi Zhang2, Yong-Chun Zhou6, Ming-Hui Yang7. 1. Department of Traditional Chinese Medicine, Chinese PLA General Hospital, Beijing, 100853, China. 2. Department of Integrated Traditional and Western Medicine of Oncology, Tangdu Hospital, the Fourth Military Medical University, Xi'an, 710032, China. 3. Department of Gastroenterology, Bethune International Peace Hospital, Shijiazhuang, 050082, China. 4. Department of Anatomy and K.K. Leung Brain Research Center, Faculty of Basic Medicine, the Fourth Military Medical University, Xi'an, 710032, China. 5. Department of Biochemistry and Molecular Biology, the Fourth Military Medical University, Xi'an, 710032, China. 6. Department of Radiotherapy Oncology, the First Affiliated Hospital, the Fourth Military Medical University, Xi'an, 710032, China. 7. Department of Traditional Chinese Medicine, Chinese PLA General Hospital, Beijing, 100853, China. ymh9651@sina.com.
Abstract
OBJECTIVE: To investigate the effects of Salvia miltiorrhiza and Ligustrazine Injection (SML) on proliferation and apoptosis of human hepatic stellate cell LX-2 and the expression of N-myc downstreamregulated gene 2 (NDRG2, a tumor suppressor gene). METHODS: HSCs from the LX-2 cell line were cultured in vitro. The proliferative state of different initial LX-2 cell numbers was measured using a 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. LX-2 cells were plated in 96-well plates at an approximate density of 2.50×104 cells/mL and cultured for 24 h followed by the application of different concentrations of SML (1, 2, 4 and 8 μL/mL). Cell proliferation was measured using the MTT assay at 24 and 48 h. Apoptosis was detected by flow cytometry at 24 h. LX-2 cells were treated with different concentrations of SML and extracted with protein lysis buffer. The levels of NDRG2 and β-catenin were measured by Western blot. RESULTS: With the exception of the 1 and 2 μL/mL concentrations, 4 and 8 μL/mL SML inhibited cell proliferation in a concentration-dependent manner at 24 and 48 h (P<0.05). With the exception of the 1 and 2 μL/mL concentrations, the NDRG2 expression level was greatly increased in a concentration-dependent manner. However, the level of β-catenin was unaffected. CONCLUSION: SML inhibit LX-2 cell proliferation in a concentration-dependent manner, and the mechanism may be associated with NDRG2 over-expression.
OBJECTIVE: To investigate the effects of Salvia miltiorrhiza and Ligustrazine Injection (SML) on proliferation and apoptosis of human hepatic stellate cell LX-2 and the expression of N-myc downstreamregulated gene 2 (NDRG2, a tumor suppressor gene). METHODS: HSCs from the LX-2 cell line were cultured in vitro. The proliferative state of different initial LX-2 cell numbers was measured using a 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. LX-2 cells were plated in 96-well plates at an approximate density of 2.50×104 cells/mL and cultured for 24 h followed by the application of different concentrations of SML (1, 2, 4 and 8 μL/mL). Cell proliferation was measured using the MTT assay at 24 and 48 h. Apoptosis was detected by flow cytometry at 24 h. LX-2 cells were treated with different concentrations of SML and extracted with protein lysis buffer. The levels of NDRG2 and β-catenin were measured by Western blot. RESULTS: With the exception of the 1 and 2 μL/mL concentrations, 4 and 8 μL/mL SML inhibited cell proliferation in a concentration-dependent manner at 24 and 48 h (P<0.05). With the exception of the 1 and 2 μL/mL concentrations, the NDRG2 expression level was greatly increased in a concentration-dependent manner. However, the level of β-catenin was unaffected. CONCLUSION: SML inhibit LX-2 cell proliferation in a concentration-dependent manner, and the mechanism may be associated with NDRG2 over-expression.
Entities:
Keywords:
N-myc downstream-regulated gene 2; Salvia miltiorrhiza and Ligustrazine Injection; apoptosis; hepatic stellate cell; proliferation
Authors: Pavel Taimr; Hajime Higuchi; Eva Kocova; Richard A Rippe; Scott Friedman; Gregory J Gores Journal: Hepatology Date: 2003-01 Impact factor: 17.425