RESUMEN
Objective To study the effect of autophagy regulation on hypoxia/reoxygenation injury(H/RI)in mouse spermatogonia,and to explore the effect of autophagy on ischemia-reperfusion injury(IRI)in mouse testicular tissue.Methods The mouse spermatogonial cell line GC1 spg was used as the research object to construct the H/RI model.Rapamycin(RAPA)and 3-methyladenine(3-MA)were used as autophagy ago-nists and inhibitors.The control group,the model group,the autophagy agonist intervention group(H/RI+autophagy agonist intervention),and the autophagy inhibitor intervention group(H/RI+autophagy inhibitor intervention)were set up.The cells proliferation ability of each group was detected by methyl thiazol tetrazoli-um(MTT)method.The release level of reactive oxygen species(ROS)and apoptosis level of each group were detected by flow cytometry.The expression levels of autophagy-related gene Beclin1 and apoptosis-relat-ed genes Bcl-2 and Bax mRNA in each group were detected by real-time fluorescence quantitative PCR(qPCR).The expression levels of autophagy-related proteins LC3-Ⅰ,LC3-Ⅱ,Beclin1,p62 and apoptosis-relat-ed proteins Bcl-2,Bax,caspase-3 in each group were detected by Western blot.Results Compared with the control group,the proliferation abiliy,the expression levels of Beclin1 and Bcl-2 mRNA in the model group were significantly decreased(P＜0.01),the relative expression levels of p62 and Bcl-2 proteins were significantly decreased(P＜0.01).The ROS level,apoptosis rate and the mRNA expression level of Bax were significantly increased(P＜0.01),and the protein expresion levels of Beclin1,Bax,caspase-3 and the protein ratio of LC3-Ⅱ/LC3-Ⅰ were significantly increased(P＜0.01).Compared with the model group,the cell proliferation a-bility,the expression levels of Beclin1 and Bcl-2 mRNA in the autophagy agonist intervention group were the protein ratio of significantly increased(P＜0.01),the protein expression levels of Beclin1 and Bcl-2 and the protein ration of LC3-Ⅱ/LC3-Ⅰ were significantly increased(P＜0.01).The ROS level,apoptosis rate and the expression level of Bax mRNA were significantly decreased(P＜0.01),and the protein expression levels of p62,Bax,caspase-3 and the protein ratio of LC3-Ⅱ/LC3-Ⅰ were significantly decreased(P＜0.01).Com-pared with the model group,the cell proliferation ability,the mRNA expression levels of Beclin1 and Bcl-2 in the autophagy inhibitor intervention group were significantly decreased(P＜0.01),the protein expression lev-els of Beclin1 and Bcl-2 protein were significantly decreased(P＜0.01).The ROS level,apoptosis rate and the mRNA expression level of Bax were significantly increased(P＜0.01),and the protein expression levels of p62,Bax,caspase-3 and the protein ratio of LC3-Ⅱ/LC3-Ⅰwere significantly iecreased(P＜0.01).Conclusion Enhanced au-tophagy can inhibit apoptosis of spermatogonia and repair H/RI in mice,which provides a theoretical basis for the treatment of testicular tissue IRI.
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Objective To investigate the effect and necessity of hysteroscopy combined with laparoscopy in the diagnosis and treatment of refractory polycystic ovarian syndrome complicated with infertility.Methods 80 cases of patients with polycystic ovary syndrome (PCOS)combined with infertility were randomly divided into two groups with 40 cases in each group.The testing group used hysteroscopy combined with laparoscopy in diagnosis and treatment while the laparoscopy was only used in the control group.The changes of ovary,ovarian follicles and serum hormone level of the two groups which before and after operation were compared,and the data of postoperative normal ovulation rate,pregnancy rate and abortion rate of the two groups were recorded.Results The follicle number and ovarian volume of the testing group were significantly decreased than that before treatment,and were significantly lower than that of the control group,with statistical difference (t =3.82,3.80,3.74,3.75,all P <0.05).The serum T,LH and LH/FSH of the testing group were significantly decreased than the data before surgery,and were significantly lower compared to the control group with statistical difference (t =9.32,4.06,3.80,all P <0.05).The normal ovulation rate,pregnancy rate and abortion rate of the testing group were significantly higher compared to the control group (90.0%,62.5% and 7.5% vs.72.5%,50% and 15%)χ2 =4.02,4.10,3.90,all P <0.05).Conclusion Hyste-roscopy combined with laparoscopy has an obvious curative effect in the diagnosis and treatment of polycystic ovary syndrome combined with infertility,which can effectively improve ovulation rate and pregnancy rate and reduce the abortion rate,is worthy of clinical application.