1.The effect and mechanism of total saponins of astragalus membranaceus combined with bone marrow mesenchymal stem cells in treating autoimmune premature ovarian failure model rats by regulating the SDF-1/CXCR4 pathway
Lina Zhao ; Chenming Zhang ; Zixue Sun ; Jianshe Chen
Acta Universitatis Medicinalis Anhui 2025;60(7):1258-1265
Objective :
To explore the effect and mechanism of total saponins of astragalus membranaceus combined with bone marrow mesenchymal stem cells (BMSCs) regulating the stromal cell-derived factor-1 (SDF-1) / chemo- kine receptor 4 ( CXCR4) pathway in the treatment of autoimmune premature ovarian failure ( POF) model rats.
Methods :
Rats were randomly divided into POF group,total saponins of astragalus membranaceus group,BMSCs group ,combination group ,combination + SDF-1 / CXCR4 pathway inhibitor ( AMD3100 ) group ,and control group,with 12 rats in each group.Except for the control group,all other groups of rats needed to construct a POF model.After successful modeling,treatment would begin.The drug injected into the tail vein once a week,and the drug injected into the abdominal cavity once a day for 4 weeks.ELISA was applied to detect the levels of anti ovari- an antibodies (AOAB) ,β-endorphin ( β-EP) ,vascular endothelial growth factor (VEGF) ,tumor necrosis factor- α (TNF-α) ,and interferon-γ ( INF-γ) in serum.Changes in ovarian index were detected.HE staining was ap- plied to detect the number of mature follicles,developing follicles,and blocked follicles in the ovary.Flow cytome- try was applied to detect the proportion of BMSCs in the ovaries.QRT-PCR was applied to detect the mRNA ex- pression of mesenchymal transition factor (cMET) ,hepatocyte growth factor (HGF) ,SDF-1,and CXCR4 in ovar- ian tissue.Western blot was applied to detect SDF-1 and CXCR4 proteins in ovarian tissue.
Results :
Compared with the control group,the serum levels of AOAB,TNF-α , INF-γ , and the number of blocked follicles in the ova- ries were higher,while the serum levels of β-EP,VEGF,ovarian index,the numbers of mature follicles and devel- oping follicles in the ovary,the expression of SDF-1,CXCR4 mRNA and protein were lower in the POF group (P < 0. 05) .Compared with the POF group,the serum levels of AOAB,TNF-α , INF-γ , and the number of blocked follicles in the ovaries were lower,while the serum levels of β-EP,VEGF,ovarian index,the numbers of mature follicles and developing follicles in the ovary,the proportion of BMSCs,the expression of cMET ,HGF,SDF-1, CXCR4 mRNA and SDF-1,CXCR4 protein were higher in the total saponins of astragalus membranaceus group, BMSCs group,and combination group,and the corresponding indicators in the combination group showed the most obvious change trend (P<0. 05) .AMD3100 reversed the effect of total saponins of astragalus membranaceus com- bined with BMSCs on POF rats.
Conclusion
The combination of total saponins of astragalus membranaceus and BMSCs may inhibit inflammation in POF rats,promote follicular development,increase the proportion of BMSCs in the ovaries,and promote their homing by activating the SDF-1 / CXCR4 pathway,thereby improving ovarian func- tion.
2.Comparative experimental study of sodium benzoate and corticosterone in a mouse model of asthenozoospermia with high sperm DNA fragmentation
Zonglin LU ; Haiyang ZHAO ; Hui WANG ; Xin WANG ; Zixue SUN
Chinese Journal of Comparative Medicine 2025;35(3):48-57
Objective To prepare a mouse model of asthenozoospermia(AZS)with high sperm DNA fragmentation(SDF)using corticosterone(CORT)and sodium benzoate(NaB).Methods Fifty 3-week-old male ICR mice were divided randomly into CORT-treated(n=30)and NaB-treated(n=20)groups.The CORT group was further divided into the following six groups(n=5 per group):high CORT(500 μg/mL),medium CORT(200 μg/mL),and low CORT(10 μg/mL)drinking water group,drinking water control group,CORT injection(40 mg/kg)group,and injection control group(normal saline).The animals were modeled continuously for 50 d.Mice in the NaB group were further divided into four groups(n=5 per group):high NaB(500 mg/kg),medium NaB(300 mg/kg),and low NaB(100 mg/kg)gavage groups,and control group(normal saline).The animals were modeled continuously for 50 d.The physiological state of the mice in each group was observed and mass changes were recorded continuously.The sperm motility capacity and DNA fragmentation index(DFI)of the sperm were observed from the tail of the epididymis after the end of the modeling.Results The rate of mass change in the CORT-injection moding group showed a downward trend.There was no significant difference(P>0.05)in the high NaB gavage group,and the rate of body mass change in the high NaB gavage group was significantly decreased compared with the control group(P<0.05).The percentages of forward motility sperm were significantly decreased in the CORT injection group(P>0.05)and the percentage in the high NaB gavage group(P<0.05),compared with the control group.The DFI was increased in the CORT injection group compared with the control group,but the difference was not significant(P>0.05),and the DFI in the high NaB gavage group was significantly increased compared with the control group(P<0.05).Conclusions Intragastric gavage with NaB 500 mg/(kg·d)for 50 d is an ideal method for constructing an animal model of AZS with high SDF.
3.Comparative experimental study of sodium benzoate and corticosterone in a mouse model of asthenozoospermia with high sperm DNA fragmentation
Zonglin LU ; Haiyang ZHAO ; Hui WANG ; Xin WANG ; Zixue SUN
Chinese Journal of Comparative Medicine 2025;35(3):48-57
Objective To prepare a mouse model of asthenozoospermia(AZS)with high sperm DNA fragmentation(SDF)using corticosterone(CORT)and sodium benzoate(NaB).Methods Fifty 3-week-old male ICR mice were divided randomly into CORT-treated(n=30)and NaB-treated(n=20)groups.The CORT group was further divided into the following six groups(n=5 per group):high CORT(500 μg/mL),medium CORT(200 μg/mL),and low CORT(10 μg/mL)drinking water group,drinking water control group,CORT injection(40 mg/kg)group,and injection control group(normal saline).The animals were modeled continuously for 50 d.Mice in the NaB group were further divided into four groups(n=5 per group):high NaB(500 mg/kg),medium NaB(300 mg/kg),and low NaB(100 mg/kg)gavage groups,and control group(normal saline).The animals were modeled continuously for 50 d.The physiological state of the mice in each group was observed and mass changes were recorded continuously.The sperm motility capacity and DNA fragmentation index(DFI)of the sperm were observed from the tail of the epididymis after the end of the modeling.Results The rate of mass change in the CORT-injection moding group showed a downward trend.There was no significant difference(P>0.05)in the high NaB gavage group,and the rate of body mass change in the high NaB gavage group was significantly decreased compared with the control group(P<0.05).The percentages of forward motility sperm were significantly decreased in the CORT injection group(P>0.05)and the percentage in the high NaB gavage group(P<0.05),compared with the control group.The DFI was increased in the CORT injection group compared with the control group,but the difference was not significant(P>0.05),and the DFI in the high NaB gavage group was significantly increased compared with the control group(P<0.05).Conclusions Intragastric gavage with NaB 500 mg/(kg·d)for 50 d is an ideal method for constructing an animal model of AZS with high SDF.
4.Clinical Observation on Herb-partitioned Moxibustion on Navel Combined with Yishen Tongluo Prescription in Treatment of Infertility Induced by Idiopathic Asthenozoospermia with Kidney-Yang Deficiency and Collateral Obstruction Syndrome
Fang ZHANG ; Peipei ZHAO ; Zixue SUN ; Jianshe CHEN ; Xiang CHEN ; Pengchao LI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(4):97-103
ObjectiveTo observe the clinical efficacy of herb-partitioned moxibustion on the navel combined with Yishen Tongluo prescription in the treatment of infertility induced by idiopathic asthenozoospermia (iAZS) with kidney-Yang deficiency and collateral obstruction syndrome and its effect on sperm DNA damage and superoxide dismutase (SOD) in the seminal plasma. MethodsA total of 112 eligible patients who met the inclusion criteria were randomly divided into an observation group (56 cases) and a control group (56 cases). The patients in the observation group were treated with herb-partitioned moxibustion on the navel combined with Yishen Tongluo prescription,while those in the control group received levocarnitine oral liquid. The primary observation indicators included spouse pregnancy rate,progressive motility (PR),and total sperm motility,and the secondary observation indicators included sperm DNA fragmentation index (DFI),SOD in the seminal plasma, and improvement of TCM syndromes. The treatment cycle was 12 weeks. Before and after treatment,the PR,total sperm motility,sperm DFI,SOD in the seminal plasma, and TCM syndrome scores were recorded. The patients were followed up for 12 weeks and the pregnancy status of spouses within 24 weeks (half a year) was recorded. The clinical efficacy of the two groups was evaluated. ResultThe pregnancy rate of spouses in the observation group was 15.69% (8/51), higher than 3.85% (2/52) in the control group (χ2=4.118,P<0.05). The total effective rate of the observation group was 88.24%(45/51), superior to 69.23% (36/52)in the control group (Z=-3.402,P<0.01). After treatment, PR, total sperm motility,sperm DFI, SOD in the seminal plasma, and TCM syndromes of the two groups were improved compared with those before treatment (P<0.05), and the observation group was superior to the control group (P<0.05). ConclusionHerb-partitioned moxibustion on the navel combined with Yishen Tongluo prescription in the treatment of iAZS-induced infertility patients with kidney-Yang deficiency and collateral obstruction syndrome can increase PR,total sperm motility, and SOD level in the seminal plasma, reduce sperm DFI,improve the TCM symptoms of patients, and improve the pregnancy rate of spouses. The mechanism may be attributed to the fact that this treatment can increase the SOD level in the seminal plasma of patients,enhance the body's antioxidant function,protect sperm from oxidative stress damage,and reduce sperm DFI.
5.Fatty acid metabolism and ovarian cancer
Tingting FAN ; Danni DING ; Zixue ZHAO ; Yang YU ; Fengjuan HAN
Practical Oncology Journal 2023;37(5):429-433
Ovarian cancer is a common malignant tumor in the female reproductive system,and its pathogenesis and regulato-ry mechanisms are extremely complex and still unclear.Fatty acid metabolism mainly involves the processes of fatty acid uptake,syn-thesis,and oxidation.Previous studies have shown that fatty acid metabolism plays a unique role in the occurrence and development of ovarian cancer.Therefore,this article reviews existing literature and delves into the correlation between fatty acid metabolism and ovarian cancer,aiming to provide new perspectives and reflections on the mechanism of fatty acid metabolism and targeted treatment of ovarian cancer.
6.Generation and epitope mapping of a monoclonal antibody against nucleoprotein of Ebola virus.
Xiaodu WANG ; Yang LIU ; Haoting WANG ; Zixue SHI ; Fanfan ZHAO ; Jianchao WEI ; Donghua SHAO ; Zhiyong MA
Chinese Journal of Biotechnology 2012;28(11):1317-1327
Ebola virus (EBOV) causes highly lethal hemorrhagic fever in humans and nonhuman primates and has a significant impact on public health. The nucleoprotein (NP) of EBOV (EBOV-NP) plays a central role in virus replication and has been used as a target molecule for disease diagnosis. In this study, we generated a monoclonal antibody (MAb) against EBOV-NP and mapped the epitope motif required for recognition by the MAb. The MAb generated via immunization of mice with prokaryotically expressed recombinant NP of the Zaire Ebola virus (ZEBOV-NP) was specific to ZEBOV-NP and able to recognize ZEBOV-NP expressed in prokaryotic and eukaryotic cells. The MAb cross-reacted with the NP of the Reston Ebola virus (REBOV), the Cote-d'Ivoire Ebola virus (CIEBOV) and the Bundibugyo Ebola virus (BEBOV) but not with the NP of the Sudan Ebola virus (SEBOV) or the Marburg virus (MARV). The minimal epitope sequence required for recognition by the MAb was the motif PPLESD, which is located between amino acid residues 583 and 588 at the C-terminus of ZEBOV-NP and well conserved among all 16 strains of ZEBOV, CIEBOV and BEBOV deposited in GenBank. The epitope motif is conserved in four out of five strains of REBOV.
Animals
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Antibodies, Monoclonal
;
immunology
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Ebolavirus
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chemistry
;
immunology
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Epitope Mapping
;
methods
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Escherichia coli
;
genetics
;
metabolism
;
Mice
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Mice, Inbred BALB C
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Nucleoproteins
;
immunology
;
Recombinant Proteins
;
biosynthesis
;
genetics
;
immunology


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