| 引用本文: | 孙凤平,马远,赵帅.Kiss-1/GPR54/GnRH及MKRN3轴观察知柏地黄汤化裁方对性早熟模型雌鼠的影响[J].中国现代应用药学,2025,42(6):80-86. |
| SUNFENGPING,MAYUAN,ZHAOSHUAI.Effect of Zhibai Dihuang Decoction on female rats with precocious puberty model was observed through KiSS-1 / GPR54 / GnRH and MKRN3 axis[J].Chin J Mod Appl Pharm(中国现代应用药学),2025,42(6):80-86. |
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| 摘要: |
| 目的 探讨知柏地黄汤化裁方对性早熟模型雌鼠MKRN3及Kiss-1/GPR54/GnRH轴的影响。方法 将雌性15日龄SD大鼠40只随机分为2组:空白组10只，模型制备组30只。模型制备组采用双酚A溶液连续灌胃，连续10天。模型鉴定成功后，随机分成3组，即模型组、中药干预组给予知柏地黄汤化裁方9.24g/kg/d、西药干预组（曲普瑞林，100ug/kg/d）。此后连续给药6周，对照组、模型组灌服等体积生理盐水；记录雌鼠阴门开启时间；采用酶联免疫吸附测定法(ELISA)检测大鼠外周血MKRN3、Kiss-1、GPR54、GnRH、LH、FSH、E2水平；采用HE染色观察卵巢子宫形态学的变化；采用qPCR和Western Blot 检查卵巢Kiss-1、GPR54、GnRH及MKRN3的mRNA和蛋白含量的表达。结果 与空白组相比较，模型组阴门开启时间较早（P<0.05）；卵巢皮质内可见各级卵泡，原始卵泡居多；子宫体积增大，可见大量腺体；卵巢、子宫组织 Kiss-1、GPR54、GnRH蛋白表达水平上调（P<0.05），MKRN3子宫、卵巢组织蛋白表达水平下调（P<0.05）；血清中相关激素Kiss-1、Kiss-1R、GnRH、LH、E2水平下降（P<0.05）；Kiss-1mRNA、GPR54mRNA、GnRHmRNA表达显著上调（P<0.05），MKRN3mRNA表达明显下调（P<0.05或P＜0.01）。与模型组比较，中药干预组阴门开启时间稍晚（P<0.05）；卵巢皮质可见各级卵泡，少量成熟卵泡；子宫体积小，腺体少见；卵巢、子宫组织 Kiss-1、GPR54、GnRHmRNA和蛋白表达水平下调（P<0.05），MKRN3子宫、卵巢组织mRNA和蛋白表达水平上调（P<0.05）；结论 知柏地黄汤化裁方可调节CCP模型雌鼠血清激素水平、抑制卵巢和子宫中的GnRH、LH、FSH、E2因子的表达、改善子宫和卵泡的发育，延缓性早熟。 |
| 关键词: 性早熟 知柏地黄汤化裁方 Kiss-1/GPR54/GnRH、MKRN3 |
| DOI：10.13748/j.cnki.issn1007-7693.20233328 |
| 分类号: |
| 基金项目:河南省中医药项目（222102310281）、河南省中医药拔尖人才培养项目（豫卫中医函2021（15）号）、河南省科技攻关联合共建项目（LHGJ20220712） |
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| Effect of Zhibai Dihuang Decoction on female rats with precocious puberty model was observed through KiSS-1 / GPR54 / GnRH and MKRN3 axis |
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SUNFENGPING1, MAYUAN2, ZHAOSHUAI2
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1.Henan Children Hospital;2.Henan Children Hospital
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| Abstract: |
| OBJECTIVE To investigate the effect of Zhibai Dihuang Tang on the MKRN3 and Kiss-1/GPR54/GnRH axes in female rats with precocious puberty. METHODS Forty 15 day old female SD rats were randomly divided into two groups: a blank group of 10 rats and a model preparation group of 30 rats. The model preparation group was continuously gavaged with bisphenol A solution for 10 days. After successful model identification, the model was randomly divided into three groups: the model group,The traditional Chinese medicine intervention group was given a modified formula of Zhibai Dihuang Tang at a dose of 9.24g/kg/d, and the western medicine intervention group (Triptorelin, 100ug/kg/d). Afterwards, the control group and model group were orally administered with an equal volume of physiological saline for 6 consecutive weeks; Record the opening time of the female mouse"s vagina; Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of MKRN3, Kiss-1, GPR54, GnRH, LH, FSH, and E2 in peripheral blood of rats; Observe the morphological changes of the ovaries and uterus using HE staining; QPCR and Western Blot were used to examine the mRNA and protein expression of ovarian Kiss-1, GPR54, GnRH, and MKRN3.. RESULTS Compared with the blank group, the model group had an earlier opening time of the vagina (P<0.05); Various levels of follicles can be seen in the ovarian cortex, with the majority being primitive follicles; The uterine volume increases, with a large number of glands visible; The protein expression levels of Kiss-1, GPR54, and GnRH in ovarian and uterine tissues were upregulated (P<0.05), while the protein expression levels of MKRN3 in uterine and ovarian tissues were downregulated (P<0.05); The levels of related hormones Kiss-1, Kiss-1R, GnRH, LH, and E2 in serum decreased (P<0.05); The expression of Kiss-1 mRNA, GPR54 mRNA, and GnRH mRNA was significantly upregulated (P<0.05), while the expression of MKRN3 mRNA was significantly downregulated (P<0.05 or P<0.01). Compared with the model group, the traditional Chinese medicine intervention group had a slightly later opening time of the vagina (P<0.05); Various levels of follicles can be seen in the ovarian cortex, with a small number of mature follicles; The uterus has a small volume and few glands; The mRNA and protein expression levels of Kiss-1, GPR54, and GnRH in ovarian and uterine tissues were downregulated (P<0.05), while the mRNA and protein expression levels of MKRN3 in uterine and ovarian tissues were upregulated (P<0.05);. CONCLUSION The modified formula of Zhibai Dihuang Tang can regulate the serum hormone levels, inhibit the expression of GnRH, LH, FSH, and E2 factors in the ovaries and uterus of CCP model female mice, improve the development of the uterus and follicles, and delay precocious puberty. |
| Key words: precocious puberty Zhi Bai Di Huang Tang Hua Fang Kiss-1/GPR54/GnRH, MKRN3 |
