木尼孜其对加入顺铂培养的大鼠卵巢颗粒细胞性激素分泌及增殖调控作用观察  

Regulatory effects of Muniziqi on sex hormone secretion and proliferation of rat ovarian granulosa cells cultured with cisplatin

作　　者：刘莉 杨银银 夏米西努尔·伊力克[1] LIU Li;YANG Yinyin;Xiamixinuer·Yilike(School of Basic Medical Sciences,Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学基础医学院,乌鲁木齐830054

出　　处：《山东医药》2022年第18期7-11,共5页Shandong Medical Journal

基　　金：新疆维吾尔自治区“十三五”重点学科资助项目(高原学科1001)。

摘　　要：目的观察木尼孜其对加入顺铂培养的大鼠颗粒细胞性激素分泌及细胞增殖的调控作用。方法①木尼孜其血清制备:60只5~6周龄雌性SD大鼠随机分为木尼孜其高、中、低剂量组,每组20只,分别用木尼孜其合剂7.6、3.8、1.9毫升/只灌胃,早晚各1次,共灌胃7 d。第7天早上将2次剂量一次性灌胃,灌胃结束1~2 h采集三组大鼠腹主动脉血,分离高、中、低剂量木尼孜其血清。②大鼠卵巢颗粒细胞制备:3周龄雌性SD大鼠10只,皮下注射孕马血清促性腺激素50 IU/只。注射48 h时处死大鼠,分离卵巢组织,鉴定卵巢颗粒细胞。③大鼠卵巢颗粒细胞分组及木尼孜其血清给予方法:取大鼠卵巢颗粒细胞,分为3组,其中1、2组加入5μg/mL的CP培养48 h。培养后1组再分为三个亚组,分别加入高、中、低剂量木尼孜其血清培养。2组为顺铂组,3组为正常对照组,均用10%胎牛血清的培养基培养。④各组大鼠卵巢颗粒细胞培养液卵泡刺激素(Follicle-stimulating hormone,FSH)、黄体生成素(Lutein⁃izing hormone,LH)、雌二醇(Estradiol 2,E2)检测:培养48 h时取各组细胞培养液,ELISA法检测细胞培养液FSH、LH和E2。⑤各组大鼠颗粒细胞增殖情况观察:分别于培养24、48、72、96 h时取各组细胞,采用CCK-8法观察细胞增殖情况。结果与正常组比较,CP组大鼠卵巢颗粒细胞培养液FSH和LH水平升高、E2水平下降(P均<0.05);与CP组比较,高剂量组大鼠卵巢颗粒细胞培养液FSH和LH水平下降、E2水平升高(P均<0.05);与低、中剂量组比较,高剂量组大鼠卵巢颗粒细胞培养液FSH和LH水平下降、E2水平升高(P均<0.05)。与正常组比较,培养24、48、72、96 h时CP组卵巢颗粒细胞OD450值均降低(P均<0.05);与CP组比较,培养24、48、72、96 h时高剂量组卵巢颗粒细胞OD450值均升高(P均<0.05);与低、中剂量组比较,培养24、48、72、96 h时高剂量组卵巢颗粒细胞OD450值均升高(P均<0.05)。结论木尼�Objective To observe the regulatory effects of Muniziqi on sex hormone secretion and cell proliferation of rat granulosa cells cultured with cisplatin.Methods①Preparation of Muniziqi serum:sixty female Sprague Dawley rats aged 5-6 weeks were randomly divided into the high-dose,medium-dose and low-dose groups(n=20).Rats in the three groups were gavaged with 7.6,3.8 and 1.9 mL of Muniziqi mixture,respectively,once in the morning and once in the evening,for a total of 7 days.On the morning of the seventh day,two doses were given by gavage at one time.At the end of 1-2 h,the abdominal aortic blood of rats in these three groups was collected,and the serum of the high-dose,medium-dose and low-dose muniziqi was separated.②Preparation of granulosa cells from rat ovary:Ten three-week-old female SD rats were subcutaneously injected with 50 IU of pregnant horse serum gonadotropin.The rats were sacrificed at 48 h after injection,and the ovarian tissues were separated and ovarian granulosa cells were identified.③Grouping of rat ovarian granulosa cells and serum administration of Muniziqi:Rat ovarian granulosa cells were taken and randomly divided into three groups.Cells in the groups 1 and 2 were cultured with 5μg/mL cisplatin for 48 h.After culture,the cells in the group 1 were further divided into three subgroups,which were cultured with high,medium and low doses of Muniziqi serum.Group 2 was the cisplatin group,group 3 was the normal control group,and all cells were cultured in medium with 10%fatal bovine serum.④Detection of follicle stimulating hormone(FSH),luteinizing hormone(LH)and estradiol 2(E2)in the ovarian granulosa cell culture medium of rats in each group:after 48 h culture,the cell culture medium of each group was taken,and FSH,LH and E2 in the cell culture fluid were detected by ELISA.⑤Observation on the proliferation of granulosa cells in each group:cells in each group were collected at 24,48,72 and 96 h,respectively,and cell proliferation was observed by CCK-8 method.Results Compared with the norma

关 键 词：木尼孜其 卵巢颗粒细胞 卵泡刺激素 黄体生成素 雌二醇 

分 类 号：R285.5[医药卫生—中药学]