CAJ | 학술논문

目的观察加味四逆散含药血清对大鼠肝星状细胞-T6 (HSC-T6)增殖与凋亡的影响.方法以不同浓度的加味四逆散含药血清[(加味四逆散低浓度组(含生药量0.78 g/ml)、加味四逆散中浓度组(含生药量1.56 g/ml)、加味四逆散高浓度组(含生药量3.12 g/ml)]作用于体外培养的HSC-T6细胞,作用时间分别为12、24、48 h后,采用噻唑蓝(MTT)比色法检测各组HSC-T6.采用流式细胞仪和TUNEL法检测对HSC-T6凋亡的影响.结果 ①加味四逆散含药血清作用于大鼠HSC-T6后,细胞增殖受到抑制,呈时间和浓度依赖性,加味四逆散各浓度组与空白对照组比较差异均有统计学意义(P＜0.01)；加味四逆散高浓度组[48 h:(0.399±0.041)％]与鳖甲软肝片组[48 h:(0.429±0.037)％]比较差异有统计学意义(P＜0.05).②流式细胞仪分析结果:与空白对照组比较,加味四逆散各浓度组、复方鳖甲软肝片组与HSC作用12、24、48 h后,细胞凋亡增加.与鳖甲软肝片组[12 h:(8.31±0.30)％,24 h:(16.25±0.25)％,48 h:(27.12±0.39)％]比较,加味四逆散中浓度组[12h: (17.83±0.25)％,24 h:(26.06±0.26)％,48h:(39.30±2.25)％]、加味四逆散高浓度组[12h:(27.15±0.29)％,24h:(38.96±0.51)％,48h:(49.34±0.77) ％]细胞凋亡均增加(P＜0.05).③TUNEL检测,与鳖甲软肝片组[(30.0±3.92) ％]比较,加味四逆散各浓度[低浓度组(25.1±1.48)％、中浓度组(39.30±2.25)％、高浓度组(39.30±2.25)％]药物血清作用48 h后,细胞凋亡率均增加(P＜0.01).结论加味四逆散含药血清可抑制体外HSC-T6的增殖、促讲其凋亡,以高剂量作用48 h最明显.
목적 관찰가미사역산함약혈청대대서간성상세포-T6 (HSC-T6)증식여조망적영향.방법 이불동농도적가미사역산함약혈청[(가미사역산저농도조(함생약량0.78 g/ml)、가미사역산중농도조(함생약량1.56 g/ml)、가미사역산고농도조(함생약량3.12 g/ml)]작용우체외배양적HSC-T6세포,작용시간분별위12、24、48 h후,채용새서람(MTT)비색법검측각조HSC-T6.채용류식세포의화TUNEL법검측대HSC-T6조망적영향.결과 ①가미사역산함약혈청작용우대서HSC-T6후,세포증식수도억제,정시간화농도의뢰성,가미사역산각농도조여공백대조조비교차이균유통계학의의(P＜0.01)；가미사역산고농도조[48 h:(0.399±0.041)％]여별갑연간편조[48 h:(0.429±0.037)％]비교차이유통계학의의(P＜0.05).②류식세포의분석결과:여공백대조조비교,가미사역산각농도조、복방별갑연간편조여HSC작용12、24、48 h후,세포조망증가.여별갑연간편조[12 h:(8.31±0.30)％,24 h:(16.25±0.25)％,48 h:(27.12±0.39)％]비교,가미사역산중농도조[12h: (17.83±0.25)％,24 h:(26.06±0.26)％,48h:(39.30±2.25)％]、가미사역산고농도조[12h:(27.15±0.29)％,24h:(38.96±0.51)％,48h:(49.34±0.77) ％]세포조망균증가(P＜0.05).③TUNEL검측,여별갑연간편조[(30.0±3.92) ％]비교,가미사역산각농도[저농도조(25.1±1.48)％、중농도조(39.30±2.25)％、고농도조(39.30±2.25)％]약물혈청작용48 h후,세포조망솔균증가(P＜0.01).결론 가미사역산함약혈청가억제체외HSC-T6적증식、촉강기조망,이고제량작용48 h최명현.
Objective To observe the effect of JWSNS serum on proliferation and apoptosis hepatic stellate cells.Methods After being added different concentrations of JWSNS (the low concentrations of JWSNS:0.78 g/ml of crude drug; the medium concentration group of JWSNS:1.56 g/ml of crude drug; the high concentration group of JWSNS:3.12 g/ml of crude drug) drug-containing serum in vitro HSC-T6 cells for 12h,24 h and 48 h respectively,detected serum HSC-T6 proliferation with MTT colorimetry method and measured HSC-T6 apoptosis with flow cytometry and TUNEL method.Results (①) After applied JWSNS on rats HSC-T6,the Cell proliferation was inhibited which showed a time-concentration dependence.The differences were significant when comparing each JWSNS group with the control group (P＜0.01).High concentration of JWSNS group showed significant difference when compared with Biejiaruangan tablets group (P＜0.05) with high concentration of JWSNS (0.399± 0.041) ％ after 48h,and Biejia-Ruangan tablets (0.429± 0.037) ％ after 48 h.② Flow cytometry analysis showed each JWSNS group and Biejiaruangan tablets group had significant increased cell apoptosis when compared with the control group (P＜0.05) after 12 h,24 h,and 48 h.JWSNS medium concentration group [12 h was (17.83±0.25)％,24 h was (26.06±0.26)％,48 h was (39.30±2.25) ％] and JWSNS high concentration group [12 h was (27.15±0.29)％,24 h was (38.96±0.51)％,48 h was (49.34± 0.77) ％] had a significant increased cell apoptosis compared to the Biejia-Ruangan tablets group [ 12 h was (8.31 ± 0.30) ％,24 h was (16.25 ± 0.25) ％,48 h was (27.12± 0.39) ％].③ TUNEL detection showed that each concentration of JWSNS group [the low concentration of JWSNS:was (25.1 ± 1.48)％,medium concentration group of JWSNS was(39.30±2.25)％,high concentration group of JWSNS was(39.30±2.25)％] had a significant increased cell apoptosis rate than Biejiaruangan tablets group (30.0± 3.92) after 48 h (P＜0.01).Conclusion JWSNS containing serum can inhibit the proliferation of HSC-T6 in vitro,promote the apoptosis