Male infertility is a significant cause of psychosocial and marital distress in approximately 50% of couples who are unable to conceive,with male factors being the underlying cause.Guijiajiao(Colla Carapacis et Plastr...Male infertility is a significant cause of psychosocial and marital distress in approximately 50% of couples who are unable to conceive,with male factors being the underlying cause.Guijiajiao(Colla Carapacis et Plastri,CCP)is a Traditional Chinese Medicine commonly used to treat male infertility.The present study aimed to investigate the potential mechanisms underlying the preventive effects of CCP on male infertility.An infertile male rat model was established using cyclophosphamide(CTX),and CCP was administered for both treatment and prevention.Fecal microbiota transplantation(FMT)was also performed to explore the role of gut microbiota in the CCP-mediated prevention of male infertility in rats.Sperm motility and concentration were determined using a semiautomatic sperm classification analyzer.Subsequently,histopathological analysis using HE staining was performed to examine the changes in the small intestine and testis.Moreover,the serum levels of lipopolysaccharide(LPS)and testosterone were measured by ELISA.In addition,immunohistochemistry was conducted to detect CD3 expression in the small intestine,while RT-qPCR was employed to assess the expressions of interleukin-1 beta(IL-1β),cluster of differentiation 3(CD3),Monocyte chemoattractant protein-1(MCP-1),and C-X-C motif chemokine ligand 10(CXCL-10)in the small intestine and epididymis.Finally,gut microbiota was analyzed by 16S rRNA sequencing.CCP improved sperm motility,number,and concentration in CTX-induced infertile male rats.CCP increased the serum testosterone level,inhibited the immune cell infiltration of the intestinal lamina propria,and promoted the aggregation of CD3+T cells in CTX-induced male infertility rats.CCP also inhibited the expressions of MCP-1,CXCL-10,and IL-1βin the epididymis of male infertility rats.At the genus level,CTX led to a reduction in the abundance of Lactobacillus,Clostridia_UCG.014,and Romboutsia in the intestinal tract of rats.In contrast,CCP decreased the abundance of Ruminococcus and increased the abundance of Romboutsia in infertile male rats.Additionally,FMT experiments proved that the gut microbiota of CCP-treated rats facilitated testicular tissue recovery and spermatogenesis while also reducing the serum LPS level in infertile male rats.CCP improves the spermatogenic ability of infertile male rats by restoring gut microbiota diversity and inhibiting epididymal inflammation.展开更多
OBJECTIVE:To investigate the effect of Guijiajiao(Colla Carapacis et Plastri, CCP), the glue of tortoise shell, on bone regeneration in zebrafish and male Wistar rats.METHODS: In this study, we applied in vitro and in...OBJECTIVE:To investigate the effect of Guijiajiao(Colla Carapacis et Plastri, CCP), the glue of tortoise shell, on bone regeneration in zebrafish and male Wistar rats.METHODS: In this study, we applied in vitro and in vivo models, tissue section analysis, and quantitative polymerase chain reaction to assess the effects of CCP on bone repair. MG-63 cells were used in alkaline phosphatase(ALP) activity and mineralization assays. Zebrafish and male Wistar rats were used to evaluate the effects of CCP on bone repair in vivo.RESULTS: A simple preparation of CCP promoted osteogenesis in vivo and in vitro, and promoted MG-63 cell proliferation as well as ALP activity and mineralization. In addition, CCP activated Akt and extracellular signaling-regulated kinase pathways and significantly increased the expression of ossification-related genes and proteins such as runt-related transcription factor-2, osteocalcin, and osteopontin. The in vivo results revealed promotion of osteogenesis by immersing zebrafish in CCP for 72 h.An oral dose of 1.25 g/kg CCP significantly improved skull defects in rats, which was accompanied by an increase in serum ALP levels.CONCLUSION: One of the ingredients of Guilu Erxianjiao(tortoise shell and deer horn formula) pro-vides a practical alternative therapy for bone regeneration.展开更多
基金Excellent Youth Project of Scientific Research Program of Hunan Education Department(No.22B0370)Project of Traditional Chinese Medicine Administration of Hunan Province(No.B2023034)Hunan Natural Science Foundation(No.2022JJ40305).
文摘Male infertility is a significant cause of psychosocial and marital distress in approximately 50% of couples who are unable to conceive,with male factors being the underlying cause.Guijiajiao(Colla Carapacis et Plastri,CCP)is a Traditional Chinese Medicine commonly used to treat male infertility.The present study aimed to investigate the potential mechanisms underlying the preventive effects of CCP on male infertility.An infertile male rat model was established using cyclophosphamide(CTX),and CCP was administered for both treatment and prevention.Fecal microbiota transplantation(FMT)was also performed to explore the role of gut microbiota in the CCP-mediated prevention of male infertility in rats.Sperm motility and concentration were determined using a semiautomatic sperm classification analyzer.Subsequently,histopathological analysis using HE staining was performed to examine the changes in the small intestine and testis.Moreover,the serum levels of lipopolysaccharide(LPS)and testosterone were measured by ELISA.In addition,immunohistochemistry was conducted to detect CD3 expression in the small intestine,while RT-qPCR was employed to assess the expressions of interleukin-1 beta(IL-1β),cluster of differentiation 3(CD3),Monocyte chemoattractant protein-1(MCP-1),and C-X-C motif chemokine ligand 10(CXCL-10)in the small intestine and epididymis.Finally,gut microbiota was analyzed by 16S rRNA sequencing.CCP improved sperm motility,number,and concentration in CTX-induced infertile male rats.CCP increased the serum testosterone level,inhibited the immune cell infiltration of the intestinal lamina propria,and promoted the aggregation of CD3+T cells in CTX-induced male infertility rats.CCP also inhibited the expressions of MCP-1,CXCL-10,and IL-1βin the epididymis of male infertility rats.At the genus level,CTX led to a reduction in the abundance of Lactobacillus,Clostridia_UCG.014,and Romboutsia in the intestinal tract of rats.In contrast,CCP decreased the abundance of Ruminococcus and increased the abundance of Romboutsia in infertile male rats.Additionally,FMT experiments proved that the gut microbiota of CCP-treated rats facilitated testicular tissue recovery and spermatogenesis while also reducing the serum LPS level in infertile male rats.CCP improves the spermatogenic ability of infertile male rats by restoring gut microbiota diversity and inhibiting epididymal inflammation.
文摘OBJECTIVE:To investigate the effect of Guijiajiao(Colla Carapacis et Plastri, CCP), the glue of tortoise shell, on bone regeneration in zebrafish and male Wistar rats.METHODS: In this study, we applied in vitro and in vivo models, tissue section analysis, and quantitative polymerase chain reaction to assess the effects of CCP on bone repair. MG-63 cells were used in alkaline phosphatase(ALP) activity and mineralization assays. Zebrafish and male Wistar rats were used to evaluate the effects of CCP on bone repair in vivo.RESULTS: A simple preparation of CCP promoted osteogenesis in vivo and in vitro, and promoted MG-63 cell proliferation as well as ALP activity and mineralization. In addition, CCP activated Akt and extracellular signaling-regulated kinase pathways and significantly increased the expression of ossification-related genes and proteins such as runt-related transcription factor-2, osteocalcin, and osteopontin. The in vivo results revealed promotion of osteogenesis by immersing zebrafish in CCP for 72 h.An oral dose of 1.25 g/kg CCP significantly improved skull defects in rats, which was accompanied by an increase in serum ALP levels.CONCLUSION: One of the ingredients of Guilu Erxianjiao(tortoise shell and deer horn formula) pro-vides a practical alternative therapy for bone regeneration.
