三甲益肝颗粒含药血清对HSC-T6细胞形态、增殖和凋亡的影响  

Effect of rat serum containing a traditional Chinese granule,San Jia Yi Gan on morphology,proliferation and apoptosis of HSC-T6 cells

作　　者：杨道坤[1] 梁海军[1] 申保生[1] 孙屹峰[1] 乔汉臣[1] 

机构地区：[1]新乡医学院第一附属医院感染疾病科,河南新乡453100

出　　处：《第三军医大学学报》2010年第5期486-489,共4页Journal of Third Military Medical University

基　　金：河南省自然科学基金(200733004)~~

摘　　要：目的观察三甲益肝颗粒(San Jia Yi Gan granula,SJG)含药血清对HSC-T6细胞形态、增殖和凋亡的作用,探讨SJG抗肝纤维化的细胞学基础。方法雄性SD大鼠30只按随机数字表法分为5组,每组6只,分别为生理盐水对照组(Control)、水飞蓟宾组(Sily)、低剂量SJG组(SJG-L)、中剂量SJG组(SJG-M)和高剂量SJG组(SJG-H)。SJG按低、中、高(1.7、3.4、6.8g/kg)3个剂量灌胃大鼠制备含药血清,并用不同体积分数(5%、10%、20%、40%)SJG含药血清干预HSC-T6细胞株,用倒置相差显微镜观察HSC-T6细胞形态学的变化,MTT法检测SJG药物血清对HSC-T6细胞增殖的影响,流式细胞仪检测SJG药物血清诱导HSC-T6细胞早期凋亡。结果含药血清干预后,细胞形态发生明显改变:药物组细胞较对照组细胞明显减少,细胞间隙较宽,细胞皱缩,核染色质浓集呈球形。MTT实验结果显示各组药物血清均可抑制HSC-T6细胞的增殖,SJG-M、SJG-H组对HSC-T6细胞增殖的抑制均显著高于Sily组(P<0.05),SJG-L组与Sily组对HSC-T6细胞增殖的抑制无显著差异(P>0.05)。SJG-M、SJG-H组凋亡率分别为(7.09±1.04)%、(11.63±1.53)%,显著高于Sily组[(4.38±1.07)%]、SJG-L组[(4.17±0.30)%]和对照组[(1.74±0.11)%]。与对照组比较,各药物血清干预组细胞凋亡数明显增加,以SJG-H组增加最为明显。结论SJG含药血清可显著抑制HSC-T6细胞的活化、增殖,并诱导其凋亡。Objective To explore the effect of the serum of rat after intragastric injection of a traditional Chinese drug, San Jia Yi Gan granula （SJG） on the morphology, proliferation and apoptosis of rat hepatic stellate cell line HSC-T6 in order to elucidate the underlying mechanism by which SJG exerts antifibrosis on hepatic cells. Methods Thirty male SD rats were randomly and equally divided into 5 groups： control group, Silybin group （Sily, 4.38 mg/kg）, low-dose SJG group （SJG-L, 1.7 g/kg）, middle-dose SJG group （SJG-M, 3.4 g/kg）, and high-dose SJG group （SJG-H, 6.8 g/kg）. Solutions of normal saline, Sily, and SJG at corresponding doses were given the rats respectively by intragastrical injection once per day, for 7 d. The rats were killed in 1 h after last injection and their heart were removed to produce the drug-contained sera. HSC-T6 cells were exposed to different doses of drug-contained serum （5%, 10%, 20% and 40% ） for 24 or 48 h. The morphology of HSC-T6 cells were observed with phase contrast microscopy, proliferation by MTF assay, and cell apoptosis by flow cytometry. Results HSC-T6 cells after the drug-contained sera treatment showed significant changes compared with those treated with the control serum, with lesser cells, intercellular space widening, shrinked cell body, and condensed chromatin to form spheres. MTT assay showed that various concentrations of drug-contained sera inhibited the proliferation of HSC-T6 cells, with SJG-M and SJG-H groups had stronger inhibitory effect than Sily group （ P 〈 0.05 ）, while whose effect was no difference with that of SJG-L group. Flow cytometry demonstrated that the apoptotic ratios of HSC-T6 cells in SJG-H group [ （ 11.63 ± 1.53 ） % ] and SJG-M group [ （7.09 ± 1.04） % ] were higher than that in SJG-L group [ （4.17 ± 0.30） % ], Sily group [ （4.38 ± 1.07 ） % ], and the control group [ （ 1.74± 0.11 ） %, P 〈 0.05 ]. SJG induced apoptosis of HSC-T6 cells in a dose-dependent manner. Conclusion SJG-contained seru

关 键 词：三甲益肝颗粒 HSC—T6细胞 细胞增殖 凋亡 肝硬化 大鼠 

分 类 号：R285.5[医药卫生—中药学] R322.47[医药卫生—中医学]