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目的研究苦参碱对胰岛素促进SD大鼠乳鼠心肌成纤维细胞(CF)增殖及胶原合成作用的影响。方法 2011年11月~2012年4月用消化法培养SD大鼠乳鼠心肌成纤维细胞(cardiac fibroblast,CF),实验分为5组:对照组只用培养液,胰岛素(Ins)组加入胰岛素(终浓度10–8mol/L),低剂量苦参碱(Mat)+Ins组0.1 mmol/L Mat+Ins,中剂量Mat+Ins组0.2 mmol/L Mat+Ins;高剂量Mat+Ins组0.4 mmol/L Mat+Ins。采用四氮唑蓝(MTT)比色法测定细胞增殖(A490值),3H-胸腺嘧啶(3H-thymidine,3H-TdR)掺入法测定CF的胶原合成功能,分别观察Ins对CF增殖和胶原合成的影响;不同浓度Mat是否可对抗Ins对CF增殖和胶原合成的作用。结果随Mat浓度的增加,CF A490值呈递减趋势,中、高剂量Mat+Ins组(0.140±0.026、0.089±0.005)明显低于对照组(0.323±0.097)(均P<0.05),低、中、高剂量Mat+Ins组的A490值均低于Ins组(P均<0.05);随Mat浓度的增加,CF的3HTdR掺入率呈递减趋势,中、高剂量Mat+Ins组(655.96±52.57、376.14±56.33)明显低于对照组(1 326.32±278.03)(P均<0.05),低、中、高剂量Mat+Ins组的3H-TdR掺入率均低于胰岛素组(P均<0.05)。结论在体外,Ins具有促进CF增殖和胶原合成的作用,而Mat则可以对抗Ins的这些作用,Mat此效应呈剂量依赖性。
Objective To investigate the effects of matrine on insulin-induced proliferation and collagen synthesis of neonatal rat cardiac fibroblasts (CFs). Methods From November 2011 to April 2012, cardiac fibroblasts (CFs) of SD rats were cultured by digestion method. The experiment was divided into five groups: the control group was fed with only the culture medium and insulin group 0.1 mmol / L Mat + Ins for the low dose Mat + Ins group, 0.2 mmol / L Mat + Ins for the medium dose Mat + Ins group, 0.4 mmol / L Mat + Ins for the high dose Mat + Ins group mmol / L Mat + Ins. Cell proliferation (A490 value) was determined by MTT colorimetric assay and the collagen synthesis function of 3H-thymidine (3H-TdR) incorporation method was measured. The effect of Ins on CF proliferation and collagen The effect of different concentrations of Mat on the proliferation and collagen synthesis of CF. Results With the increase of Mat concentration, the value of CF A490 showed a decreasing trend. The levels of CF A490 in Mat + Ins group (0.140 ± 0.026,0.089 ± 0.005) were significantly lower than those in control group (0.323 ± 0.097) (P <0.05) The A490 values of medium and high dose Mat + Ins group were lower than those of Ins group (all P <0.05). With the increase of Mat concentration, the incorporation rate of 3HTdR in CF group showed a decreasing trend. The medium and high dose Mat + Ins group (655.96 ± 52.57,376.14 ± 56.33) was significantly lower than that of the control group (1 326.32 ± 278.03) (P <0.05). The 3H-TdR incorporation rates of low, medium and high dose Mat + Ins groups were lower than that of the insulin group (P < 0.05). Conclusion In vitro, Ins has the effect of promoting CF proliferation and collagen synthesis, whereas Mat can antagonize these effects of Ins, and this effect is dose-dependent.