青春发育期5-羟色胺合成下降对雌鼠青春期启动的影响

doi:10.3969/j.issn.1674-8115.2022.03.007

摘要/Abstract

摘要：

目的·初步探讨青春发育期下丘脑5-羟色胺（5-hydroxytryptamine，5-HT）合成下降对雌鼠青春期启动的影响。方法·将72只21日龄SD雌性大鼠随机分为氯苯丙氨酸（para-chlorophenylalanine，PCPA）组和生理盐水组（对照组），每组36只。每日向PCPA组大鼠腹腔注射PCPA（以建立5-HT合成下降模型），对照组则注射等量生理盐水，给药处理均持续至处死前1 d。分别在28日龄、35日龄和42日龄处死大鼠，采用实时荧光定量PCR（quantitative real-time PCR，qPCR）检测2组大鼠下丘脑促性腺激素释放激素（gonadotropin-releasing hormone，GnRH）基因的表达，采用酶联免疫吸附试验（enzyme linked immunosorbent assay，ELISA）检测2组大鼠下丘脑组织中5-HT含量、GnRH含量以及血清中黄体生成激素（luteinizing hormone，LH）水平。同时，从28日龄开始，每日9：00~10：00观察并记录2组大鼠阴门开放时间。结果·5-HT含量的ELISA结果显示，对照组大鼠在28日龄最低、42日龄最高，整体呈递增趋势，与PCPA组的趋势相一致；同对照组相比，PCPA组大鼠在上述3个时间点的5-HT含量均较低（均P<0.05）。GnRH含量的qPCR及ELISA结果显示，与对照组相比，PCPA组大鼠在上述3个时间点的GnRH mRNA相对表达量、GnRH含量均较低（均P<0.05）。LH水平的ELISA结果显示，同对照组相比，PCPA组大鼠在28日龄、35日龄有所下降且差异具有统计学意义（均P<0.05）。阴门开放时间的观察结果显示，与对照组相比，PCPA组大鼠的阴门开放时间有所延迟（P=0.035）。结论·青春发育期5-HT合成下降可致SD雌性大鼠下丘脑组织中GnRH合成减少、血清LH水平下降、阴门开放延后，从而推迟了其青春期启动。

关键词: 5-羟色胺, 大鼠, 青春发育期, 促性腺激素释放激素

Abstract:

Objective·To preliminarily explore the effects of decreased 5-hydroxytryptamine (5-HT) synthesis in the hypothalamus during puberty development on the onset of puberty in female rats.

Methods·Seventy two 21-day-old female SD rats were randomly divided into para-chlorophenylalanine (PCPA) group and normal saline group (control group), with 36 rats in each group. The rats in the PCPA group were injected with PCPA intraperitoneally every day (to establish the model of decreased 5-HT synthesis), while the rats in the control group were injected with the same amount of normal saline. The administration treatment lasted until 1 d before death. The rats were sacrificed at the age of 28 days, 35 days and 42 days, respectively. The expression of gonadotropin-releasing hormone (GnRH) gene in the hypothalamus of the two groups was detected by quantitative real-time PCR (qPCR). The content of 5-HT and GnRH in the hypothalamus and the level of luteinizing hormone (LH) in peripheral blood were detected by enzyme-linked immunosorbent assay (ELISA). At the same time, since the age of 28 days, the vaginal opening time in the two groups was observed and recorded from 9:00 to 10:00.

Results·The ELISA results of 5-HT content showed that the rats in the control group had the lowest level at 28 days and the highest level at 42 days, and showed an overall increasing trend, which was consistent with the trend of the PCPA group. Compared with the control group, the content of 5-HT in the PCPA group was lower at the above three time points (all P<0.05). qPCR and ELISA results of GnRH content showed that compared with the control group, the expressions of GnRH mRNA and GnRH protein in the PCPA group were lower at the above three time points (all P<0.05). The results of ELISA showed that compared with the control group, LH levels of the PCPA group were lower at the age of 28 days and 35 days, and the difference was statistically significant (both P<0.05). The observation results showed that compared with the control group, vaginal opening time of the PCPA group was delayed (P=0.035).

Conclusion·The decrease of 5-HT synthesis during puberty development can lead to decrease of GnRH synthesis in the hypothalamus, decrease of LH level in peripheral blood and delay of vaginal opening, thus delaying the onset of puberty in female SD rats.

Key words: 5-hydroxytryptamine (5-HT), rat, puberty development, gonadotropin-releasing hormone (GnRH)

中图分类号:

R725.8

董婷, 尹小琴, 李妍, 李嫔. 青春发育期5-羟色胺合成下降对雌鼠青春期启动的影响[J]. 上海交通大学学报（医学版）, 2022, 42(3): 307-311.

DONG Ting, YIN Xiaoqin, LI Yan, LI Pin. Effect of decreased 5⁃hydroxytryptamine synthesis during puberty development on the onset of puberty in female rats[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(3): 307-311.

图/表 5

表1 qPCR引物序列

Tab 1 Primer sequences for qPCR

Gene

Forward sequence （5' →3'）

Reverse sequence

（5' →3'）

Length/bp

GnRH

CCGCTGTTGTTCTGTTGACTGTG

GGGGTTCTGCCATTTGATCCTC

150

β-actin

GTCAGGTCATCACTATCGGCAAT

AGAGGTCTTTACGGATGTCAACGT

147

图 1 2组大鼠下丘脑中5-HT含量分析Note： ①P=0.000, ②P=0.005, ③P=0.009, compared with the control group.

Fig 1 Analysis of 5-HT content in the hypothalamus of the two groups

图2 2组大鼠下丘脑中 GnRH mRNA及GnRH蛋白的表达Note： A. Relative expression of GnRH mRNA detected by qPCR. B. Expression of GnRH protein detected by ELISA. ①P=0.000, ②P=0.004, ③P=0.009, ④P=0.001, compared with the control group.

Fig 2 Expression of GnRH mRNA and GnRH protein in the hypothalamus of the two groups

图3 2组大鼠的血清LH水平分析Note： ①P=0.020, ②P=0.000, compared with the control group.

Fig 3 Analysis of LH levels in serum of the two groups

图4 2组大鼠阴门开放时间分析

Fig 4 Analysis of vaginal opening time in the two groups

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