补肾阴及补肾阳法对化疗诱导的卵巢早衰大鼠外周血TNF－α、IFN－γ水平及卵巢颗粒细胞凋亡的影响

摘要

AIM:To treat the chemotherapy-induced premature ovarian failure ( POF) rats with the methods of tonifying kidney yin and yang by observing the indicator changes in apoptosis of ovarian granulosa cells and related cyto -kines before and after medication .METHODS:SD rats ( n=36) were chronically given intraperitoneal injection of cyclo-phosphamide ( CTX) for 15 d in order to establish the animal model of POF .The rats were randomly divided into control group, CTX group, CTX+kidney yin group and CTX +kidney yang group.The levels of tumor necrosis factor alpha (TNF-α) and interferon gamma ( IFN-γ) in the peripheral blood were detected by ELISA .The apoptosis of ovarian granulosa cells was observed by TUNEL method .The levels of apoptosis related proteins were determined by Western blot .RE-SULTS:After modeling , the levels of TNF-αand IFN-γin the peripheral blood were significantly increased .The protein expression of Bax was up-regulated and Bcl-2 was down-regulated .The apoptosis of ovarian granulosa cells was significantly induced with a deep staining .After treatment with tonifying kidney yin and yang , the levels of TNF-αand IFN-γin the pe-ripheral blood were decreased , the protein expression of Bax was down-regulated and Bcl-2 was up-regulated, and the apop-tosis of ovarian granulosa cells was reduced .CONCLUSION:The reserve function of the ovarian can be recovered by pre-scriptions of tonifying the kidney yin and yang via regulating the expression of TNF -α, IFN-γ, Bax and Bcl-2, and inhibi-ting the apoptosis of ovarian granulosa cells .%目的：观察化疗诱导的卵巢早衰大鼠接受补肾阴法和补肾阳法干预后，其外周血肿瘤坏死因子－α（TNF－α）、干扰素－γ（IFN－γ）水平及卵巢颗粒细胞凋亡情况。方法：48只雌性SD大鼠随机分成对照（control）组、环磷酰胺（CTX）组、环磷酰胺＋补肾阴（CTX＋kidney yin）组、环磷酰胺＋补肾阳（CTX＋kidney yang）组。 ELISA法检测各组大鼠外周血TNF－α、IFN－γ水平；末端脱氧核糖核苷酸转移酶介导的缺口末端标记（TUNEL）法检测卵巢颗粒细胞凋亡；Western blot法检测卵巢Bax、Bcl－2蛋白的表达。结果： CTX组大鼠外周血TNF－α、IFN－γ表达水平上升，卵巢颗粒细胞凋亡明显，卵巢中Bax蛋白明显增多，而Bcl－2蛋白明显减少。经本实验中补肾方药治疗后，外周血TNF－α、IFN－γ水平下降，颗粒细胞凋亡程度减轻，Bax蛋白减少，Bcl－2蛋白增多，部分指标变化以CTX＋kidney yin组较为明显。结论：补肾阴及补肾阳法可通过调控颗粒细胞凋亡相关的Bax、Bcl－2蛋白及外周血TNF－α、IFN－γ的水平，抑制卵巢颗粒细胞凋亡，恢复卵巢功能、增强储备能力。