疏肝益阳胶囊对炔雌醚诱导大鼠生精紊乱和氧化应激的缓解作用
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摘要
笔者拟探讨疏肝益阳胶囊(SGYY)对炔雌醚(QES)诱导大鼠生精紊乱和氧化应激的缓解作用及机制。将20只8周龄雄性SD大鼠随机分为4组,适应饲养1周后,进行药物灌胃处理,对照组:0.1mL生理盐水+0.1mL橄榄油;SGYY组:100 mg·kg-1 SGYY;QES组:0.1 mg·kg-1 QES;QES+SGYY组:0.1 mg·kg-1 QES+100mg·kg-1 SGYY。QES溶于橄榄油;SGYY溶于生理盐水;每天1次,连续2周。处理结束后,取睾丸、附睾、精囊腺和前列腺称重、测量长径与短径,并制备睾丸组织切片,通过HE染色,观察睾丸生精小管组织结构、生精细胞比例的变化;通过免疫组织化学方法检测睾丸生精细胞PCNA表达,Tunel法检测细胞凋亡;分离血浆,检测睾酮含量变化。取睾丸匀浆检测抗氧化酶活性。结果显示,QES处理后大鼠生殖器官的质量显著下降,附睾的精子数量显著减少;生精小管的面积、直径、生精上皮的高度和生精细胞数量均显著减少。而且,生精细胞的PCNA表达和血浆睾酮含量明显下降,Tunel阳性细胞数明显增加。睾丸SOD、GSH-Px和T-AOC活性显著下降,MDA含量显著升高。SGYY增加生殖器官质量、精子数量、睾丸睾酮含量、增殖的生精细胞数量和抗氧化酶活性,降低了MDA含量和Tunel的表达。结果表明,SGYY通过改善抗氧化功能、抑制氧化应激及促进睾酮分泌等途径增加了生精细胞数量并提高睾丸的生精功能。
The study was aimed to evaluate the probable protective effect of Shugan Yiyang Capsule(SGYY,Chinese herbal extract with highly efficient antioxidant activities)against testicular toxicity and oxidative stress in adult male rats exposed to quinestrol(QES)and study the possible mechanisms underlying these effects.Twenty 8-week-old male SD rats were randomly divided into 4 groups.After 1 week of adaptation,the rats were treated with gavage once daily for 2 weeks.Control group:0.1 mL physiological saline+0.1 mL olive oil;SGYY group:100 mg·kg-1 SGYY;QES group:0.1 mg·kg-1 QES;QES+SGYY group:0.1 mg·kg-1 QES+100 mg·kg-1 SGYY.QES was dissolved in olive oil;SGYY was dissolved in physiological saline.After the treatments,the testis,epididymis,seminal vesicle and prostate were weighed,and the long and short diameters were measured.The changes in the histological structure of seminiferous tubules and the proportion of spermatogenic cells were observed in testicular tissue sections by HE staining.The expression of PCNA in spermatogenic cells of testis was detected by immunohistochemistry,and the apoptosis was detected by Tunel.The blood plasma was separated and the content of testosterone was detected.The antioxidant enzyme activity was detected in the testicular homogenate.The results showed that exposure to QES decreased the weights of reproductive organs and the sperms counts in cauda epididymis of rats.A significant decrease in the seminiferous tubular area,tubular diameter,height of germinal epithelium,number of spermatogenic cell(per seminiferous tubules)and expression of PCNA level with a significant increase in the Tunel levels were observed in the testis of experimental rats over the controls.These events were accompanied by a significant reduction in the plasma testosterone levels in QES exposed rats,indicating reduced steroidogenesis.Significant reduction in the activities of SOD,GSH-Px and T-AOC and a significant increase in MDA were also observed in the testis of QES exposed rats over the controls.Conversely,supplementation of SGYY(100 mg·kg-1)ameliorated the male reproductive health in rats exposed to QES as evidenced by increased reproductive organ weights,sperm counts,testicular steroidogenesis,spermatogenic cell proliferation and testicular antioxidants whereas MDA and Tunel levels decreased.To sum up,the results show that SGYY increased the number of spermatogenic cells and improved spermatogenesis in the testis by improving antioxidant function,inhibiting oxidative stress and promoting testosterone secretion.
The study was aimed to evaluate the probable protective effect of Shugan Yiyang Capsule(SGYY,Chinese herbal extract with highly efficient antioxidant activities)against testicular toxicity and oxidative stress in adult male rats exposed to quinestrol(QES)and study the possible mechanisms underlying these effects.Twenty 8-week-old male SD rats were randomly divided into 4 groups.After 1 week of adaptation,the rats were treated with gavage once daily for 2 weeks.Control group:0.1 mL physiological saline+0.1 mL olive oil;SGYY group:100 mg·kg-1 SGYY;QES group:0.1 mg·kg-1 QES;QES+SGYY group:0.1 mg·kg-1 QES+100 mg·kg-1 SGYY.QES was dissolved in olive oil;SGYY was dissolved in physiological saline.After the treatments,the testis,epididymis,seminal vesicle and prostate were weighed,and the long and short diameters were measured.The changes in the histological structure of seminiferous tubules and the proportion of spermatogenic cells were observed in testicular tissue sections by HE staining.The expression of PCNA in spermatogenic cells of testis was detected by immunohistochemistry,and the apoptosis was detected by Tunel.The blood plasma was separated and the content of testosterone was detected.The antioxidant enzyme activity was detected in the testicular homogenate.The results showed that exposure to QES decreased the weights of reproductive organs and the sperms counts in cauda epididymis of rats.A significant decrease in the seminiferous tubular area,tubular diameter,height of germinal epithelium,number of spermatogenic cell(per seminiferous tubules)and expression of PCNA level with a significant increase in the Tunel levels were observed in the testis of experimental rats over the controls.These events were accompanied by a significant reduction in the plasma testosterone levels in QES exposed rats,indicating reduced steroidogenesis.Significant reduction in the activities of SOD,GSH-Px and T-AOC and a significant increase in MDA were also observed in the testis of QES exposed rats over the controls.Conversely,supplementation of SGYY(100 mg·kg-1)ameliorated the male reproductive health in rats exposed to QES as evidenced by increased reproductive organ weights,sperm counts,testicular steroidogenesis,spermatogenic cell proliferation and testicular antioxidants whereas MDA and Tunel levels decreased.To sum up,the results show that SGYY increased the number of spermatogenic cells and improved spermatogenesis in the testis by improving antioxidant function,inhibiting oxidative stress and promoting testosterone secretion.
引文
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[18]KOPALLI S R,CHA K M,JEONG M S,et al.Pectinase-treated Panax ginseng ameliorates hydrogen peroxide-induced oxidative stress in GC-2sperm cells and modulates testicular gene expression in aged rats[J].J Ginseng Res,2016,40(2):185-195.
[19]莫秋柏,乔飞.疏肝益阳胶囊治疗低睾酮血症所致ED的临床效果观察[J].临床合理用药杂志,2016,9(7):88-89.MO Q B,QIAO F.Clinical observation of Shugan Yiyang capsule in the treatment of ED caused by low testosterone[J].Chinese Journal of Clinical Rational Drug Use,2016,9(7):88-89.(in Chinese)
[20]张会波,尤建军,杜宏宏,等.疏肝益阳胶囊联合万艾可对糖尿病男性性功能障碍患者血清睾酮、雌二醇水平及性功能的影响[J].现代生物医学进展,2017,17(28):5512-5515.ZHANG H B,YOU J J,DU H H,et al.Effect of Shugan Yiyang capsule combined with viagra on serum testosterone and estradiol levels and sexual function of male diabetic patients with sexual dysfunction[J].Progress in Modern Biomedicine,2017,17(28):5512-5515.(in Chinese)
[21]谭文举,韩雯雯,闫良.疏肝益阳胶囊治疗精液液化异常的临床观察[J].中成药,2018,40(5):1227-1230.TAN W J,HAN W W,YAN L.Clinical observation on treatment of abnormal semen liquefaction by Shugan Yiyang capsule[J].Chinese Traditional Patent Medicine,2018,40(5):1227-1230.(in Chinese)
[2]LEWIN A,ARBUCKLE T E,FISHER M,et al.Univariate predictors of maternal concentrations of environmental chemicals:the MIREC study[J].Int JHyg Environ Health,2017,220(2):77-85.
[3]LI J,WANG H W,ZHANG J L,et al.Abnormal secretion of reproductive hormones and antioxidant status involved in quinestrol-induced reproductive toxicity in adult male rat[J].Tissue Cell,2014,46(1):27-32.
[4]SINGH J N,JEHAN Q,SETTY B S,et al.Effect of some steroids on spermatogenesis&fertility of rats[J].Indian J Exp Biol,1971,9(2):132-137.
[5]DIPASQUALE G,WELA P,NEMITH P.Comparison of the prolonged estrogenic and anti-fertility properties of quinestrol and four analogues[J].Steroids Lipids Res,1974,5(1):57-64.
[6]ZHAO M R,LIU M,LI D,et al.Anti-fertility effect of levonorgestrel and quinestrol in Brandt's voles(Lasiopodomys brandtii)[J].Integr Zool,2007,2(4):260-268.
[7]LV X H,GUO Y,SHI D.Effects of quinestrol on reproductive hormone expression,secretion,and receptor levels in female Mongolian gerbils(Meriones unguiculatus)[J].Theriogenology,2012,77(6):1223-1231.
[8]WU M H,LI J L,HE X X,et al.Radiation removal of synthetic estrogens in aqueous solution:influence of reduction or oxidation system and toxicity test[J].Nucl Sci Tech,2016,27(1):22.
[9]SHEN W,SHI D Z,WANG D,et al.Inhibitive effects of quinestrol on male testes in Mongolian gerbils(Meriones unguiculatus)[J].Res Vet Sci,2012,93(2):907-913.
[10]GENG Q Q,LI P L,ZHANG W B,et al.The bioaccumulation and biotransformation of synthetic estrogen quinestrol in crucian carp[J].Aquat Toxicol,2014,155:84-90.
[11]CHEN Y,SU Q Q,LIU Q S.Effects of quinestrol on the vocal behavior of mice during courtship interactions[J].Physiol Behav,2017,173:216-222.
[12]陆海旺,林思伟,王杰,等.劲男春颗粒对老龄大鼠阴茎海绵体组织氧化应激损伤的保护作用[J].中医药导报,2016,22(14):24-26.LU H W,LIN S W,WANG J,et al.Protective effects of Jinnanchun granules on oxidative stress in cavernous tissue of aged rats[J].Guiding Journal of Traditional Chinese Medicine and Pharmacy,2016,22(14):24-26.(in Chinese)
[13]王琦,倪平,吴卫平,等.疏肝益阳胶囊治疗勃起功能障碍的作用机理研究[J].中国中药杂志,2005,30(1):58-63.WANG Q,NI P,WU W P,et al.Study of Shugan Yiyang capsule on erectile dysfunction[J].China Journal of Chinese Materia Medica,2005,30(1):58-63.(in Chinese)
[14]王济,白明华,刘保兴,等.疏肝益阳胶囊对动脉性勃起功能障碍大鼠性功能及性腺质量的作用[J].安徽中医学院学报,2013,32(2):54-57.WANG J,BAI M H,LIU B X,et al.Effect of Shugan Yiyang capsules on sexual function and sex gland weight in rats with arteriogenic erectile dysfunction[J].Journal of Anhui Traditional Chinese Medical College,2013,32(2):54-57.(in Chinese)
[15]ZHANG Y H,SONG M Y,RUI X L,et al.Supplemental dietary phytosterin protects against 4-nitrophenol-induced oxidative stress and apoptosis in rat testes[J].Toxicol Rep,2015,2:664-676.
[16]AHMED E,NAGAOKA K,FAYEZ M,et al.Suppressive effects of long-term exposure to P-nitrophenol on gonadal development,hormonal profile with disruption of tissue integrity,and activation of caspase-3in male Japanese quail(Coturnix japonica)[J].Environ Sci Pollut Res,2015,22(14):10930-10942.
[17]ABDELBAKI S A,SABRY J H,AL-ADL A M,et al.The impact of coexisting sperm DNA fragmentation and seminal oxidative stress on the outcome of varicocelectomy in infertile patients:a prospective controlled study[J].Arab J Urol,2017,15(2):131-139.
[18]KOPALLI S R,CHA K M,JEONG M S,et al.Pectinase-treated Panax ginseng ameliorates hydrogen peroxide-induced oxidative stress in GC-2sperm cells and modulates testicular gene expression in aged rats[J].J Ginseng Res,2016,40(2):185-195.
[19]莫秋柏,乔飞.疏肝益阳胶囊治疗低睾酮血症所致ED的临床效果观察[J].临床合理用药杂志,2016,9(7):88-89.MO Q B,QIAO F.Clinical observation of Shugan Yiyang capsule in the treatment of ED caused by low testosterone[J].Chinese Journal of Clinical Rational Drug Use,2016,9(7):88-89.(in Chinese)
[20]张会波,尤建军,杜宏宏,等.疏肝益阳胶囊联合万艾可对糖尿病男性性功能障碍患者血清睾酮、雌二醇水平及性功能的影响[J].现代生物医学进展,2017,17(28):5512-5515.ZHANG H B,YOU J J,DU H H,et al.Effect of Shugan Yiyang capsule combined with viagra on serum testosterone and estradiol levels and sexual function of male diabetic patients with sexual dysfunction[J].Progress in Modern Biomedicine,2017,17(28):5512-5515.(in Chinese)
[21]谭文举,韩雯雯,闫良.疏肝益阳胶囊治疗精液液化异常的临床观察[J].中成药,2018,40(5):1227-1230.TAN W J,HAN W W,YAN L.Clinical observation on treatment of abnormal semen liquefaction by Shugan Yiyang capsule[J].Chinese Traditional Patent Medicine,2018,40(5):1227-1230.(in Chinese)