苦参碱对胰岛素促乳鼠心肌成纤维细胞增殖及胶原合成的影响

Effects of matrine on insulin to promote cardiac fibroblastproliferation and collagen synthesis in rats

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（cardiacfibroblast，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0．1mmol／LMat＋Ins，中剂量Mat＋Ins组0．2mmol／LMat＋Ins；高剂量Mat＋Ins组0．4mmol／LMat＋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，cFA枷值呈递减趋势，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。H—TdR掺人率呈递减趋势，中、高剂量Mat＋Ins组（655．96±52．57、376．14±56．33）明显低于对照组（1326．32±278．03）（P均〈0．05），低、中、高剂量Mat＋Ins组的3H—TdR掺人率均低于胰岛素组（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 to promote cardiac fibrohlast （CF）proliferation and collagen synthesis in rats. Methods From November 2011 to April 2012, digestion method was used to culture cardiac fi- hmblasts of SD rats. There were five groups in this experiment：①control group, managed with culture fluid; ②insulin group, with insulin（final concentration 10 -8mol·-1） ;③low dose Mat ＋ Ins group, with 0. 1mmol·L-1 Mat ＋ Ins; ④ middle dose Mat ＋ Ins group, with 0.2mmol·L-1 Mat ＋ Ins;⑤high dose Mat ＋ Ins group, with 0.4mmol·L-1 Mat ＋ Ins. CF proliferation was measured by thiazolyl blue（MTr） assay（ A490）, and collagen synthesis by 3H - thymidine（3H - TdR） uptake test. The effects of matrine on insulin to promote CF proliferation and collagen synthesis were observed, as well as the action of different concentrations of Mat against CF proliferation and collagen synthesis. Results As Mat concentration increased, CF A490 value showed a decreasing trend. It was （0. 140±0.026 ） and （0.089±0.005） respectively in middle and high dose Mat ＋ Ins groups, which were significantly lower than that in control group（0.323±0.097, P 〈 0.05 ） . A490 value in low, middle and high dose Mat ＋ Ins groups was significantly lower than that in Ins group（P 〈 0.05 ）. With the increasing of Mat concemration, the rate of 3H - TdR incorporation into CF showed a decreasing trend. It was （655.96±52.57 ） and （376.14±56.33 ） respectively in middle and high dose Mat ＋ Ins groups, which were signifi- cantly lower than that in control group（ 1 326.32±278.03, P 〈 0.05 ）. The rate of 3H -TdR incorporation into CF was lower in low, middle and high dose Mat ＋ Ins groups than in Ins group（P 〈 0.05 ）. Conclusions Insulin can promote CF proliferation and collagen synthesis in vitro, which can be inhibited by Mat. This action of Mat is dose - dependent.