Although spermatozoa are formed during spermatogenesis in the testis, testicular spermatozoa are immature and cannot swim or fertilize. These critical spermatozoal functions are acquired in the epididymis where a spec...Although spermatozoa are formed during spermatogenesis in the testis, testicular spermatozoa are immature and cannot swim or fertilize. These critical spermatozoal functions are acquired in the epididymis where a specific luminal environment is created by the blood-epididymal barrier; proteins secreted by epididymal principal cells bind to maturing spermatozoa and regulate the maturational process of the spermatozoa. In the epididymis, epithelial cell-cell interactions are mediated by adhering junctions, necessary for cell adhesion, and by tight junctions, which form the blood-epididymal barrier. The regulation of these cellular junctions is thought to represent a key determinant in the process of sperm maturation within the epididymis. Tight junctions between adjacent principal cells permit the formation of a specific microenvironment in the lumen of the epididymis that is essential for sperm maturation. Although we have made significant progress in understanding epididymal function and the blood-epididymal barrier, using animal models, there is limited information on the human epididymis. If we are to understand the normal and pathological conditions attributable to human epididymal function, we must clearly establish the physiological, cellular and molecular regulation of the human epididymis, develop tools to characterize these functions and develop clinical strategies that will use epididymal functions to improve treatment of infertility. （Asian J Androl 2007 July; 9： 463- 475）展开更多
The difference between human normal and carcinoma lung cells was studied with regard to the protein expression level and localization of the cadherin/catenin/actin complex. Results demonstrated that normal lung cell R...The difference between human normal and carcinoma lung cells was studied with regard to the protein expression level and localization of the cadherin/catenin/actin complex. Results demonstrated that normal lung cell RF expressed high levels of N-cadherin, β-catenin, α-catenin. These 3 proteins were colocalized at AJs and their submembrane adhesion plaques where they link the Rho-phalloidin-positive actin stress fibers, indicating the existence of N-cadherin/catenin/actin complexes at the AJs. Aberrant expression of AJ proteins and the actin cytoskelecton in carcinoma PG cells was observed: (1) inhibition of N-cadherin and to a degree of inhibition of α-catenin protein expression; (2) varied protein modification of β-catenin in cytoplasm soluble fraction and altered distribution of immunofluorescence: majorly in the cytoplasm and minorly on the membrane; (3) disassembly of actin stress fibers and formation of actin bodies in the cytoplasm. The data suggest that inhibited expression of AJ proteins is correlated with the disruption of the AJ complexes and the actin cytoskeleton in carcinoma PG cells, and responsible for its metastasis behaviors.展开更多
OBJECTIVES:To investigate the therapeutic effect of Huluan decotion(护卵汤,HLD)on cyclophosphamideinduced premature ovarian failure(POF)in mice and its regulatory mechanisms.METHODS:Female BALB/c mice were administere...OBJECTIVES:To investigate the therapeutic effect of Huluan decotion(护卵汤,HLD)on cyclophosphamideinduced premature ovarian failure(POF)in mice and its regulatory mechanisms.METHODS:Female BALB/c mice were administered cyclophosphamide and administered received different doses of HLD for 28 d.Levels of sex hormone,such as estradiol(E2),follicle stimulating hormone(FSH)and luteinizing hormone(LH)in the sera,were assessed using enzyme-linked immunosorbent assay(ELISA).Follicular structure variances were observed through hematoxylin and eosin(HE)staining,while Forkhead box L2(FOXL2)expression were analyzed via immuneohistochemical staining.The primary mechanism of POF were investigated through Western blot analysis.RESULTS:E2 levels decreased,and FSH and LH levels increased in POF model mice,but these trends were reversed with HLD or premarin administration,the expressions of WNT family member 4(Wnt4),β-Catenin and FOXL2 were downregulated in POF model mice,whereas high expression levels were observed in control mice and other groups.CONCLUSION:HLD effectively treats POF induced with cyclophosphamide in mice by enhancing expressions of Wnt4,β-Catenin and FOXL2.展开更多
Sclerosteosis,an ultra-rare disorder characterised by high bone mass(HBM)and skeletal overgrowth,leads to facial paralysis,hearing loss and raised intracranial pressure,which is currently managed only through high-ris...Sclerosteosis,an ultra-rare disorder characterised by high bone mass(HBM)and skeletal overgrowth,leads to facial paralysis,hearing loss and raised intracranial pressure,which is currently managed only through high-risk surgery.Sclerosteosis is caused by SOST mutations and loss of functional sclerostin,a protein that suppresses osteogenesis by antagonising Wnt/β-catenin signalling.Herein,using in vitro and in vivo approaches,we explore whether LGK974,another potent Wnt inhibitor that targets porcupine(PORCN,Wnt-specific acyltransferase),is a promising sclerosteosis therapeutic.In vitro assays showed that 100 nmol/L LGK974 significantly reduced osteoblast alkaline phosphatase(ALP)activity/mineralisation,decreased Wnt/osteoblast marker(Axin2,Runx2 and Ocn)expression,and downregulated ossification and the Wnt signalling pathway,without affecting osteoclast numbers/resorption.To assess in vivo effects,6-week-old male and female Sost deficient(Sost-/-)mice received LGK974 for 4 weeks and right hindlimbs were subjected to 20 N peak loading to assess mechanoadaptive interactions.µCT revealed significant reductions in vertebral trabecular number and lower cortical bone volume in loaded and non-loaded tibiae in male and female LGK974-treated Sost-/-mice.Interestingly,the target engagement biomarker Axin2 was only significantly reduced in male vertebrae,which may indicate differences in male and female response to LGK974.This study also shows that PORCN inhibition may effectively limit characteristic HBM and skeletal overgrowth in sclerosteosis patients at sites with severe pathology.展开更多
OBJECTIVE:To explore the therapeutic potential of the Dujieqing(DJQ)decoction(毒结清复方)for multiple myeloma(MM)and elucidate its mechanism of action.METHODS:RPMI8226 cells were treated with DJQcontaining serum(DJQ-C...OBJECTIVE:To explore the therapeutic potential of the Dujieqing(DJQ)decoction(毒结清复方)for multiple myeloma(MM)and elucidate its mechanism of action.METHODS:RPMI8226 cells were treated with DJQcontaining serum(DJQ-CS)and a Wnt/β-catenin pathway inhibitor,XAV-939.Cell counting kit-8 assay was used to examine cell viability,and flow cytometry was performed to examine apoptosis.Real-time polymerase chain reaction and Western blotting were used to evaluate the Wnt/β-catenin pathway family members in the cells.Subsequently,the RPMI8226 cells were subcutaneously injected into the left flank of none obesity disease and server combined immune-deficiency mice to replicate the xenograft tumor mouse models,which were treated with the DJQ decoction for 14 d.Hematoxylin and eosin staining was used to examine the pathological changes of the liver and kidney tissues,and to detect xenograft tumors.Wnt/β-catenin pathway family members were evaluated via Western blotting.RESULTS:DJQ-CS significantly reduced the m RNA and protein expression levels ofβ-catenin,c-myc,cyclin D1,and lymphoid enhancer binding factor 1(LEF1)while inhibiting the proliferation of RPMI8226 cells and inducing their apoptosis.Similar results were observed when the Wnt/β-catenin pathway was suppressed by inhibitors.Moreover,in the mouse model of xenograft tumors,DJQ decoction not only reduced the tumor volume but also inhibited the protein levels ofβ-catenin,c-myc,cyclin D1,and LEF1.The histopathology of the mice also showed increased apoptosis in tumor tissues,while the DJQ decoction treatment did not cause any pathological damage to the kidneys or liver.CONCLUSION:Our results indicate that the DJQ decoction suppresses tumor progression by inhibiting the Wnt/β-catenin pathway,offering a promising treatment approach for MM.展开更多
OBJECTIVE:To determine the mechanism of electroacupuncture(EA)effect by the wingless-related integration site(Wnt)/β-catenin pathway in the guinea pig myopia model.METHODS:Following myopia induction and EA,guinea pig...OBJECTIVE:To determine the mechanism of electroacupuncture(EA)effect by the wingless-related integration site(Wnt)/β-catenin pathway in the guinea pig myopia model.METHODS:Following myopia induction and EA,guinea pigs were treated with biometry to evaluate refraction and axial length.Hematoxylin and eosin(HE)staining was used to observe that the retina,choroid,and sclera had abnormal morphology.At 4,6,and 8 weeks,quantitative polymerase chain reaction(q PCR)was used to identify the expression of matrix metallopeptidase-2(MMP-2)/MMP-3/tissue inhibitor of metalloprotease-2(TIMP-2)/TIMP-3/Wnt family member 2B(WNT2B)/WNT3A/WNT7B/beta-catenin 1(CTNNB1),and dickkopf wnt signaling pathway inhibitor 1(DKK-1)m RNAs in the retina,choroid,and sclera.Western blot was used to detect the protein expression of WNT7B/2B/3A,CTNNB1 and DKK-1 in retina,choroid and sclera at 4 weeks.Enzyme-linked immunosorbent assay was used to detect the protein expression of MMP-2/TIMP-2 and MMP-3/TIMP-3 in serum at 4 weeks.Moreover,a DKK-1 inhibitor was injected into the vitreous cavity,and the expression of the above molecules was detected.RESULTS:EA could reduce the optic axial length and diopter and ameliorate ocular pathology,inhibited the expression of MMP-2/MMP-3 and WNT2B/WNT3A/WNT7B/CTNNB1,while increased the expression levels of TIMP-2/TIMP-3 and DKK-1.However,the expression levels of WNT2B/WNT3A/WNT7B/CTNNB1 and MMP-2/MMP-3 were significantly increased,and the TIMP-2/TIMP-3 and DKK-1 expression levels were decreased after injected DKK-1 inhibitor.CONCLUSION:The mechanism of EA's effects on myopia may involve the downregulation of the Wnt/β-catenin pathway and correct MMP-2/MMP-3/TIMP-2/TIMP-3 balance.展开更多
文摘Although spermatozoa are formed during spermatogenesis in the testis, testicular spermatozoa are immature and cannot swim or fertilize. These critical spermatozoal functions are acquired in the epididymis where a specific luminal environment is created by the blood-epididymal barrier; proteins secreted by epididymal principal cells bind to maturing spermatozoa and regulate the maturational process of the spermatozoa. In the epididymis, epithelial cell-cell interactions are mediated by adhering junctions, necessary for cell adhesion, and by tight junctions, which form the blood-epididymal barrier. The regulation of these cellular junctions is thought to represent a key determinant in the process of sperm maturation within the epididymis. Tight junctions between adjacent principal cells permit the formation of a specific microenvironment in the lumen of the epididymis that is essential for sperm maturation. Although we have made significant progress in understanding epididymal function and the blood-epididymal barrier, using animal models, there is limited information on the human epididymis. If we are to understand the normal and pathological conditions attributable to human epididymal function, we must clearly establish the physiological, cellular and molecular regulation of the human epididymis, develop tools to characterize these functions and develop clinical strategies that will use epididymal functions to improve treatment of infertility. （Asian J Androl 2007 July; 9： 463- 475）
文摘The difference between human normal and carcinoma lung cells was studied with regard to the protein expression level and localization of the cadherin/catenin/actin complex. Results demonstrated that normal lung cell RF expressed high levels of N-cadherin, β-catenin, α-catenin. These 3 proteins were colocalized at AJs and their submembrane adhesion plaques where they link the Rho-phalloidin-positive actin stress fibers, indicating the existence of N-cadherin/catenin/actin complexes at the AJs. Aberrant expression of AJ proteins and the actin cytoskelecton in carcinoma PG cells was observed: (1) inhibition of N-cadherin and to a degree of inhibition of α-catenin protein expression; (2) varied protein modification of β-catenin in cytoplasm soluble fraction and altered distribution of immunofluorescence: majorly in the cytoplasm and minorly on the membrane; (3) disassembly of actin stress fibers and formation of actin bodies in the cytoplasm. The data suggest that inhibited expression of AJ proteins is correlated with the disruption of the AJ complexes and the actin cytoskeleton in carcinoma PG cells, and responsible for its metastasis behaviors.
基金Fujian Natural Science:Study on Potential Protein Targets of Huluan Decotion in the Intervention of Premature Ovarian Failure(No.2021J011173)Major Project Cultivation Plan Project of Ningde Normal University:the Effect of Huluan Decotion on the Decreased Ovarian Reserve Function Induced by Cyclophosphamide is Studied based on Forkhead box L2(No.2019ZDK06)。
文摘OBJECTIVES:To investigate the therapeutic effect of Huluan decotion(护卵汤,HLD)on cyclophosphamideinduced premature ovarian failure(POF)in mice and its regulatory mechanisms.METHODS:Female BALB/c mice were administered cyclophosphamide and administered received different doses of HLD for 28 d.Levels of sex hormone,such as estradiol(E2),follicle stimulating hormone(FSH)and luteinizing hormone(LH)in the sera,were assessed using enzyme-linked immunosorbent assay(ELISA).Follicular structure variances were observed through hematoxylin and eosin(HE)staining,while Forkhead box L2(FOXL2)expression were analyzed via immuneohistochemical staining.The primary mechanism of POF were investigated through Western blot analysis.RESULTS:E2 levels decreased,and FSH and LH levels increased in POF model mice,but these trends were reversed with HLD or premarin administration,the expressions of WNT family member 4(Wnt4),β-Catenin and FOXL2 were downregulated in POF model mice,whereas high expression levels were observed in control mice and other groups.CONCLUSION:HLD effectively treats POF induced with cyclophosphamide in mice by enhancing expressions of Wnt4,β-Catenin and FOXL2.
文摘Sclerosteosis,an ultra-rare disorder characterised by high bone mass(HBM)and skeletal overgrowth,leads to facial paralysis,hearing loss and raised intracranial pressure,which is currently managed only through high-risk surgery.Sclerosteosis is caused by SOST mutations and loss of functional sclerostin,a protein that suppresses osteogenesis by antagonising Wnt/β-catenin signalling.Herein,using in vitro and in vivo approaches,we explore whether LGK974,another potent Wnt inhibitor that targets porcupine(PORCN,Wnt-specific acyltransferase),is a promising sclerosteosis therapeutic.In vitro assays showed that 100 nmol/L LGK974 significantly reduced osteoblast alkaline phosphatase(ALP)activity/mineralisation,decreased Wnt/osteoblast marker(Axin2,Runx2 and Ocn)expression,and downregulated ossification and the Wnt signalling pathway,without affecting osteoclast numbers/resorption.To assess in vivo effects,6-week-old male and female Sost deficient(Sost-/-)mice received LGK974 for 4 weeks and right hindlimbs were subjected to 20 N peak loading to assess mechanoadaptive interactions.µCT revealed significant reductions in vertebral trabecular number and lower cortical bone volume in loaded and non-loaded tibiae in male and female LGK974-treated Sost-/-mice.Interestingly,the target engagement biomarker Axin2 was only significantly reduced in male vertebrae,which may indicate differences in male and female response to LGK974.This study also shows that PORCN inhibition may effectively limit characteristic HBM and skeletal overgrowth in sclerosteosis patients at sites with severe pathology.
基金Supported by National Science Foundation-Funded Project:based on the Connection Sclerostin with Wnt/β-catenin Pathway to Explored the Effect Mechanism of Dujieqing Oral Liquid on the Inhibit Tumor and Promote Bone Formation in Multiple Myeloma(No.81960839)Innovation Project of Guangxi Graduate Education-Funded Project:Sclerostin Regulates Wnt/β-catenin Pathway to Explore the Mechanism of Dujieqing Decoction in Inhibiting Tumor Growth and Promoting Osteogenesis of Multiple Myeloma(No.YCSW2021224)To Investigate the Effect and Mechanism of Dujiangqing Oral Liquid on Osteogenic Differentiation of Mesenchymal Stem Cells in Multiple Myeloma Mice based on Wnt/β-catenin Signaling Pathway(No.YCSW2023386)。
文摘OBJECTIVE:To explore the therapeutic potential of the Dujieqing(DJQ)decoction(毒结清复方)for multiple myeloma(MM)and elucidate its mechanism of action.METHODS:RPMI8226 cells were treated with DJQcontaining serum(DJQ-CS)and a Wnt/β-catenin pathway inhibitor,XAV-939.Cell counting kit-8 assay was used to examine cell viability,and flow cytometry was performed to examine apoptosis.Real-time polymerase chain reaction and Western blotting were used to evaluate the Wnt/β-catenin pathway family members in the cells.Subsequently,the RPMI8226 cells were subcutaneously injected into the left flank of none obesity disease and server combined immune-deficiency mice to replicate the xenograft tumor mouse models,which were treated with the DJQ decoction for 14 d.Hematoxylin and eosin staining was used to examine the pathological changes of the liver and kidney tissues,and to detect xenograft tumors.Wnt/β-catenin pathway family members were evaluated via Western blotting.RESULTS:DJQ-CS significantly reduced the m RNA and protein expression levels ofβ-catenin,c-myc,cyclin D1,and lymphoid enhancer binding factor 1(LEF1)while inhibiting the proliferation of RPMI8226 cells and inducing their apoptosis.Similar results were observed when the Wnt/β-catenin pathway was suppressed by inhibitors.Moreover,in the mouse model of xenograft tumors,DJQ decoction not only reduced the tumor volume but also inhibited the protein levels ofβ-catenin,c-myc,cyclin D1,and LEF1.The histopathology of the mice also showed increased apoptosis in tumor tissues,while the DJQ decoction treatment did not cause any pathological damage to the kidneys or liver.CONCLUSION:Our results indicate that the DJQ decoction suppresses tumor progression by inhibiting the Wnt/β-catenin pathway,offering a promising treatment approach for MM.
基金Supported by the National Natural Youth Science Foundation:Research on the Mechanism of Acupuncture in Modulating the Inflammatory Microenvironment via the Muscarinic Acetylcholine Receptor(mAChR1)Signaling Pathway for Myopia Intervention(82205198)China Postdoctoral Science Foundation Project:Research on the Mechanism of Acupuncture in Modulating the Inflammatory Microenvironment via the mAChR1 Signaling Pathway for Myopia Intervention(2022M711984)+5 种基金Shandong Provincial Natural Science Foundation General Program:to Investigate the Underlying Mechanism of Acupuncture in Treating Myopia Associated eith Kidney Yang Deficiency Syndrome through the Muscarinic Acetylcholine Receptors(M-AChRs)Signaling Pathway(ZR2020MH393)Postdoctoral Innovation Project of Shandong Province:Research on the Mechanism of Electroacupuncture in Treating Myopia with"Kidney Yang Deficiency Syndrome"through the M-AChRs/Gamma-Aminobutyric Acid Signaling Pathway(202101012)China Postdoctoral Foundation General Program:to Investigate the Underlying Mechanism of Acupuncture in Treating Myopia Associated with Kidney Yang Deficiency Syndrome through the M-AChRs Signaling Pathway(2020M672127)National Natural Science Foundation:Research on the Influence of Acupuncture on the Accommodative Function and Visual Cortex Functional Network of Myopia and its Mechanism(82074498)National Key R&D Project:National Key Research and Development Program"Basic Research on High Myopia"(2019YFC1710200)National Natural Science Foundation of China:Research on the Relationship between Lens-Induced Myopia based on Omics Technology and Traditional Chinese Medicine Syndrome Types and its Molecular Mechanism(82474579)。
文摘OBJECTIVE:To determine the mechanism of electroacupuncture(EA)effect by the wingless-related integration site(Wnt)/β-catenin pathway in the guinea pig myopia model.METHODS:Following myopia induction and EA,guinea pigs were treated with biometry to evaluate refraction and axial length.Hematoxylin and eosin(HE)staining was used to observe that the retina,choroid,and sclera had abnormal morphology.At 4,6,and 8 weeks,quantitative polymerase chain reaction(q PCR)was used to identify the expression of matrix metallopeptidase-2(MMP-2)/MMP-3/tissue inhibitor of metalloprotease-2(TIMP-2)/TIMP-3/Wnt family member 2B(WNT2B)/WNT3A/WNT7B/beta-catenin 1(CTNNB1),and dickkopf wnt signaling pathway inhibitor 1(DKK-1)m RNAs in the retina,choroid,and sclera.Western blot was used to detect the protein expression of WNT7B/2B/3A,CTNNB1 and DKK-1 in retina,choroid and sclera at 4 weeks.Enzyme-linked immunosorbent assay was used to detect the protein expression of MMP-2/TIMP-2 and MMP-3/TIMP-3 in serum at 4 weeks.Moreover,a DKK-1 inhibitor was injected into the vitreous cavity,and the expression of the above molecules was detected.RESULTS:EA could reduce the optic axial length and diopter and ameliorate ocular pathology,inhibited the expression of MMP-2/MMP-3 and WNT2B/WNT3A/WNT7B/CTNNB1,while increased the expression levels of TIMP-2/TIMP-3 and DKK-1.However,the expression levels of WNT2B/WNT3A/WNT7B/CTNNB1 and MMP-2/MMP-3 were significantly increased,and the TIMP-2/TIMP-3 and DKK-1 expression levels were decreased after injected DKK-1 inhibitor.CONCLUSION:The mechanism of EA's effects on myopia may involve the downregulation of the Wnt/β-catenin pathway and correct MMP-2/MMP-3/TIMP-2/TIMP-3 balance.
