里氏木霉发酵提取黑豆异黄酮及其对雌性大鼠卵巢早期生长反应基因-1表达的影响

Black Bean Isoflavones Extracted by Trichoderma rimannii Fermentation and Its Effects on Ovarian Early Growth Response Gene-1 Expression of Female Rats

本试验旨在研究里氏木霉发酵法提取黑豆异黄酮的发酵条件,以及黑豆异黄酮对雌性大鼠卵巢早期生长反应基因-1(EGR-1)表达的影响。采用单因素试验和正交试验,研究里氏木霉发酵提取黑豆异黄酮的适宜发酵条件。选取40只49日龄、体重(200±20)g的SD青年雌性大鼠作为青年组,40只12月龄、体重(490±20)g的SD老年雌性大鼠作为老年组,2组大鼠各分为5组,每组8只,分为空白组(NC组)、阳性对照组(PC组)及低(L组)、中(M组)、高剂量黑豆异黄酮组(H组)。L、M和H组分别灌胃黑豆异黄酮100、200、300 mg/(kg BW·d),PC组灌胃己烯雌酚0.5 mg/(kg BW·d),持续30 d。结果显示,里氏木霉发酵提取黑豆异黄酮的适宜条件为发酵时间48 h、瓶装比例1∶10、菌液与豆粉的比例7∶1,此时的异黄酮提取率最佳,约为12.06%。给与黑豆异黄酮干预后,M、H和PC组的EGR-1蛋白质和mRNA表达水平相比于NC组均显著或极显著提高(P<0.05或P<0.01);并且老年组与青年组之间比较,EGR-1蛋白质表达水平在L、M组差异极显著(P<0.01),EGR-1 mRNA表达水平在L、H组差异显著或极显著(P<0.05或P<0.01)。由此可见,黑豆异黄酮作为一种植物雌激素,一定剂量下可以提高EGR-1蛋白质和mRNA的表达水平,效果类似于雌激素。

The purpose of this study was to investigate the fermentation conditions of black soybean isoflavones extracted by Trichoderma reesei fermentation,and the effects of black soybean isoflavones on expression of early growth response gene-1(EGR-1)in ovarian of female rats.The single factor test and orthogonal test were used to study the suitable fermentation conditions of black soybean isoflavones extracted by Trichoderma reesei fermentation.Forty young SD female rats at 49days of age with body weight of(200±20)g were selected as the youth group,and forty elderly SD female rats at 12 months of age with body weight of(490±20)g were selected as the elderly group,rats in youth group and elderly group were each divided into 5 groups with 8 rats per group,which were blank group(NC group),positive control group(PC group)and low(L group),medium(M group),high dose black bean isoflavones groups(H group).Rats in L,M and H groups were given 100,200 and 300 mg/(kg BW·d)black bean isoflavone by gavage,and rats in the PC group were given 0.5 mg/(kg BW·d)diethylstilbestrol by gavage,lasted for 30 days.The results showed that the optimal conditions for the black bean isoflavones extracted by Trichoderma reesei fermentation were fermentation time of 48 h,bottled ratio of 1∶10,the ratio of bacterial liquid to soybean powder of 7∶1,and the best extraction rate was about 12.06%.After the treatment with black bean isoflavones,compared with the NC group,the expression levels of EGR-1 protein and mRNA in M,H and PC groups were significantly increased(P<0.05 or P<0.01).Compared between youth group and elderly group,the expression level of EGR-1 protein in L and M groups was significantly difference(P<0.01),and the expression levels of EGR-1 gene in L and H groups was significantly difference(P<0.05 or P<0.01).In conclusion,as a phytoestrogens,black bean isoflavones can increase the expression levels of EGR-1 protein and mRNA at a certain dose,and the effect is similar to estrogen.