[關(guān)鍵詞]
[摘要]
目的 探究麒麟丸治療少弱精子癥(AS)的作用機(jī)制。方法 SD大鼠隨機(jī)分為對照組、模型組、左卡尼?。栃运?,100 mg∙kg−1)和麒麟丸低、高劑量(0.5、1.5 mg∙kg−1)組,除對照組外,其余4組ig給予雷公藤多苷35 mg∙kg−1 7 d誘導(dǎo)AS模型,從造模開始即給藥,每天1次,共ig給藥21 d。稱量睪丸和附睪組織質(zhì)量,計算大鼠生殖器官指數(shù);通過精子參數(shù)檢測系統(tǒng)檢測各組動物的精子質(zhì)量和精子形態(tài);ELISA測定大鼠血清中性激素含量;HE染色觀察給藥后各組大鼠睪丸組織病理學(xué)改變,并統(tǒng)計生精細(xì)胞和間質(zhì)細(xì)胞數(shù)量;通過免疫熒光染色檢測大鼠睪丸組織中支持細(xì)胞數(shù)量;Western blotting和實時熒光定量PCR(qRT-PCR)法檢測細(xì)胞色素P450家族19亞家族A成員1(CYP19A1)、細(xì)胞色素P450家族11亞家族A成員1(CYP11A1)和雌激素受體α(ESR1)的蛋白和基因表達(dá)水平。結(jié)果 與對照組相比,模型組精子質(zhì)量和血清睪酮(T)、雌二醇(E2)含量顯著降低(P<0.01),血清黃體生成素(LH)含量顯著升高(P<0.01),生殖器官指數(shù)和睪丸生精細(xì)胞、間質(zhì)細(xì)胞和支持細(xì)胞數(shù)量顯著降低(P<0.01),睪丸組織性激素合成限速酶CYP11A1和CYP19A1、ESR1的基因和蛋白顯著下調(diào)(P<0.01);與模型組相比,麒麟丸高劑量組精子質(zhì)量和血清T、E2含量顯著增加(P<0.01),血清LH含量顯著降低(P<0.01),生殖器官指數(shù)、睪丸生精細(xì)胞、間質(zhì)細(xì)胞和支持細(xì)胞數(shù)量顯著升高(P<0.01),CYP11A1和CYP19A1、ESR1基因和蛋白顯著上調(diào)(P<0.01)。結(jié)論 麒麟丸可通過上調(diào)性激素合成限速酶CYP11A1和CYP19A1,上調(diào)體內(nèi)性激素表達(dá)治療AS。
[Key word]
[Abstract]
Objective To explore the mechanism of Qilin Pills in the treatment of Asthenospermia (AS). Methods SD rats were randomly divided into a control group, a model group, levocarnitine (positive drug, 100 mg∙kg−1), and low and high dose (0.5, 1.5 mg∙kg−1) groups of Qilin Pills. Except for the control group, the other four groups were induced with tripterygium glycosides at a dose of 35 mg∙kg−1 for 17 days, starting from the modeling stage and administered once a day for a total of 21 days. The reproductive organ index of rats was calculated by weighing testes and epididymis. The sperm quality and morphology of each group were detected by the sperm parameter detection system. The content of sex hormone in serum was determined by ELISA. The histopathological changes of testis were observed by H&E staining, and the number of spermatogenic cells and interstitial cells were counted. The number of sertoli cells in testicular tissue was detected by immunofluorescence staining. Western blotting and real-time fluorescence quantitative PCR (qRT-PCR) were used to detect the protein and gene expression levels of cytochrome P450 family 19 subfamily A member 1(CYP19A1), cytochrome P450 family 11 subfamily A member 1(CYP11A1) and estrogen receptor α (ESR1). Results Compared with control group, the sperm quality and serum testosterone and estradiol contents of AS rats were significantly decreased (P < 0.01), the serum luteinizing hormone content was significantly increased (P < 0.01), and the number of reproductive organ index, testicular spermatogenic cells, interstitial cells and sertoli cells were significantly decreased (P < 0.01). The genes and proteins of the rate-limiting enzymes CYP11A1 and CYP19A1 and estrogen receptor ESR1 in testicular tissue were significantly down-regulated (P < 0.01). Compared with model group, the sperm quality and serum testosterone and estradiol contents in high dose of Qilin Pills group were significantly increased (P < 0.01), the serum luteinogen content was significantly decreased (P < 0.01), and the number of reproductive organ index, testicular spermatogenic cells, interstitial cells and sertoli cells were significantly increased (P < 0.01). The genes and proteins of the ring-limiting enzymes CYP11A1 and CYP19A1 and estrogen receptor ESR1 in testicular tissue were significantly up-regulated (P < 0.01).Conclusion Qilin Pills can be used to treat asthenospermia by up-regulating sex hormone expression of the rate-limiting enzymes CYP11A1 and CYP19A1.
[中圖分類號]
R285.5
[基金項目]
江蘇省高校青藍(lán)工程優(yōu)秀教學(xué)團(tuán)隊項目(2021年度);國家衛(wèi)生健康委計劃生育藥具不良反應(yīng)監(jiān)測中心開放課題(JSHD202323)