资癸益冲方对卵巢储备功能下降大鼠卵巢组织铁代谢及凋亡的影响

Effects of Zigui Yichong Formula on iron metabolism and cell apoptosis in ovarian tissue of rats with diminished ovarian reserve

目的 探讨资癸益冲方对卵巢储备功能下降大鼠卵巢组织铁代谢调节蛋白表达及细胞凋亡的影响。方法 将动情周期正常的大鼠随机分为正常组、模型组、资癸益冲方组和辅酶Q10组,每组10只,采用腹腔注射环磷酰胺(CTX)法建立卵巢储备功能下降大鼠模型,各组灌胃给予相应药物。ELISA法检测血清性激素水平,HE染色观察卵巢组织各级卵泡数量,普鲁士蓝染色观察卵巢组织铁沉积,TUNEL法检测卵巢组织细胞凋亡,Western blot法检测卵巢组织铁代谢调节蛋白转铁蛋白受体1(TFR1)、二价金属离子转运蛋白1(DMT1)及膜铁转运蛋白1(FPN1)的表达。结果 资癸益冲方可改善卵巢储备功能下降大鼠血清性激素水平,促进卵巢组织各级卵泡生长,减少卵泡闭锁,减轻卵巢组织铁沉积及细胞凋亡程度,降低卵巢组织铁代谢调节蛋白TFR1、DMT1蛋白表达,增强FPN1蛋白表达(P<0.05)。结论 资癸益冲方可通过影响卵巢储备功能下降大鼠卵巢组织铁代谢调节蛋白的表达,减轻铁沉积和细胞凋亡,进而改善卵巢储备功能。

AIM To investigate the effects of Zigui Yichong Formula on the regulations of protein expression of iron metabolism and cell apoptosis in ovarian tissue of rats with diminished ovarian reserve.METHODS Rats with normal estrous cycle were randomly divided into the normal group, the model group, Zigui Yichong Formula group and coenzyme Q10group, with 10 rats in each group. The rat model of diminished ovarian reserve was established by intraperitoneal injection of cyclophosphamide(CTX). The rats in each group given corresponding drugs by gavage then had their serum sex hormone levels detected by ELISA;their number of follicles at each level in the ovarian tissue observed by HE staining;their iron deposition in the ovarian tissue observed by Prussian blue staining;their cell apoptosis in the ovarian tissue detected by TUNEL method;and their expressions of iron metabolism regulatory protein transferrin receptor 1(TFR1), divalent metal ion transporter 1(DMT1), and ferroportin 1(FPN1) in the ovarian tissue detected by Western blot.RESULTS In addition to its efficacy in improving the rat serum hormone levels, Zigui Yichong Formula contributed also to the promoted follicles growth at all levels, reduced follicular atresia, decreased degree of iron deposition and cell apoptosis, lowered expressions of TFR1 and DMT1 proteins, and enhanced expression of FPN1 protein(P<0.05)in ovarian tissue of rats with diminished ovarian reserve.CONCLUSION Zigui Yichong Formula can improve ovarian reserve function in rats with diminished ovarian reserve through reduction of iron deposition and cell apoptosis by affecting the regulatory protein expression of iron metabolism.