隔药灸脐对卵巢早衰模型大鼠Bcl—2、Bax蛋白和mRNA表达的影响
2017-07-13卢岩田梦沈庆思张晶孙振高张永臣贾红玲
卢岩 田梦 沈庆思 张晶 孙振高 张永臣 贾红玲



摘要:目的 觀察隔药灸脐对卵巢早衰模型大鼠Bcl-2和Bax表达的影响,探讨其对卵巢的保护机制。方法 采用腹腔注射环磷酰胺建立大鼠卵巢早衰模型。48只雌性Wistar大鼠随机分为正常组、模型组、隔药灸脐组、西药组,每组12只。隔药灸脐组大鼠隔药灸“神阙”,30 min/d,隔日1次,连续14 d;西药组大鼠予戊酸雌二醇灌胃,每日1次,连续14 d。采用免疫组化、Western blot和实时荧光定量PCR检测大鼠卵巢凋亡相关因子Bcl-2、Bax蛋白和mRNA的表达。结果 模型组大鼠卵巢质量明显低于正常组(P<0.05)。Bcl-2、Bax蛋白主要表达在细胞胞浆中,隔药灸脐组和西药组较模型组卵泡颗粒细胞Bax表达范围明显缩小,Bcl-2表达范围明显增加。与正常组比较,模型组大鼠卵巢Bcl-2蛋白和mRNA表达明显降低(P<0.05,P<0.01),Bax蛋白和mRNA表达显著升高(P<0.05,P<0.01);与模型组比较,隔药灸脐组和西药组大鼠卵巢Bcl-2蛋白和mRNA表达显著升高(P<0.05,P<0.01),Bax蛋白和mRNA表达明显降低(P<0.05,P<0.01)。结论 隔药灸脐可能通过上调卵巢早衰模型大鼠卵巢Bcl-2和下调Bax的表达,改善卵巢功能。
关键词:卵巢早衰;隔药灸脐;神阙;Bcl-2蛋白;Bax蛋白;大鼠
DOI:10.3969/j.issn.1005-5304.2017.07.010
中图分类号:R245 文献标识码:A 文章编号:1005-5304(2017)07-0040-05
Abstract: Objective To observe the effects of herb-partition moxibustion at navel on the expressions of Bcl-2 and Bax in rats with premature ovarian failure (POF); To explore its mechanism underlying improvement of POF. Methods POF model was established by intraperitoneal injection of cyclophosphamide. Forty-eight Female Wistar rats were randomly divided into normal group, model group, moxibustion group and Western medicine group, with 12 rats in each group. Rats of moxibustion group were treated with herb-partition moxibustion at navel (Shenque, CV8), 30 min/d, once every other day; rats of Western medicine group were given intragastric dose of estradiol valerate, once a day, for 14 d. The protein and mRNA expressions of Bax and Bcl-2 in ovarian granulosa cells were detected by immunohistochemal method, Western blot and qRT-PCR, respectively. Results The weights of ovaries in the model group rats were significantly lower than those in the normal group (P<0.05). The expressions of Bcl-2 and Bax protein in ovarian tissue of each group were mainly expressed brown coloration and in the cytoplasm of cells. The range of Bax protein in granulosa cells of moxibustion group and Western medicine group was significantly less than that in
model group. The range of Bcl-2 in moxibustion group and Western medicine group was significantly larger than that in model group. Compared with the normal group, the expressions of Bcl-2 protein and mRNA in the model group rats significantly decreased (P<0.05, P<0.01), and the expression of Bax protein and mRNA significantly increased (P<0.05, P<0.01). The expression level of Bcl-2 protein and mRNA was significantly higher in the moxibustion group and in the western medicine group than that in the model group (P<0.05, P<0.01). The expression level of Bax protein and mRNA in the moxibustion group was significantly lower than that in the model group (P<0.05, P<0.01). Conclusion The herb-partition moxibustion at navel can improve the function of ovaries by up regulating the expression of Bcl-2 and down regulating the expression of Bax in POF rats model.
Key words: premature ovarian failure; herb-partition moxibustion at navel; Shenque (CV8); Bcl-2; Bax; rats
卵巢早衰(premature ovarian failure,POF)是指女性40岁前出现持续性闭经、性器官萎缩、高卵泡刺激素、黄体生成素水平及低雌激素水平等围绝经期症状。流行病学研究显示,POF发病率在30岁前约为1/1000[1],40岁前约为1%~3.6%[2],发病率呈逐年上升趋势。隔药灸脐法综合艾灸、药物、穴位三者的优势,能激发、调动和增强机体自身的调节系统,可促进子宫、卵巢的血液循环,从而促进卵泡的发育和成熟。卵巢颗粒细胞在卵泡发育过程中起关键作用,颗粒细胞功能的减退、凋亡可触发卵泡闭锁,是卵巢早衰的根本原因。Bax、Bcl-2为调节卵巢颗粒细胞的凋亡重要因子。Bcl-2蛋白能抑制细胞凋亡,促进卵泡生长、发育;Bax蛋白则促进细胞凋亡,使卵泡停止发育,发生闭锁[3-4],导致卵巢早衰。本实验观察隔药灸脐对卵巢早衰大鼠卵巢组织Bcl-2、Bax蛋白和mRNA表达的影响,探讨其治疗机制。
1 材料与方法……p>
