Wen-lin Gong1,Chuang Sha2,Gang Du1,Zhong-gui Shan3,Zhong-quan Qi3,Su-fang Zhou1,Nuo Yang1,4,Yong-xiang Zhao1,4.First published:21 June 2017;10(5):454-460.DOI:10.1016/j.apjtm.2017.05.004 The authors would like to corre...Wen-lin Gong1,Chuang Sha2,Gang Du1,Zhong-gui Shan3,Zhong-quan Qi3,Su-fang Zhou1,Nuo Yang1,4,Yong-xiang Zhao1,4.First published:21 June 2017;10(5):454-460.DOI:10.1016/j.apjtm.2017.05.004 The authors would like to correct an error in Figure 3 in which the flow cytometric scattergram of CD4/CD44 for the control group was erroneously used for the scattergram of CD8/CD44 for the PVIDSC group.The correct scattergram of CD8/CD44 for the PVIDSC group is provided below.The error does not affect the conclusion of the study.The authors apologize for the error and the inconvenience it might have caused to readers.展开更多
Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups.Each bullfrog in bacterium-injected group was injected intraperitoneally(i.p.) w...Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups.Each bullfrog in bacterium-injected group was injected intraperitoneally(i.p.) with 0.2 ml bacterial suspension at a density of 5.2 × 106 CFU/ml,while each one in control group injected i.p.with 0.2 ml sterile saline solution(0.85%,w/v).Three bullfrogs in both groups were sampled at 0,1,3,7,11,15 and 20 days post-injection(dpi) for the evaluation of non-specific immune parameters.It was observed that intraperitoneal injection of A.hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood.Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group.However,a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes.No significant difference in changes in the number of peripheral erythrocytes,serum superoxide dismutase(SOD) activity,and lysozyme activity was detected between the two groups.It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A.hydrophila infection.展开更多
The model of acute lung injury(ALI)was established by intraperitoneal administration,but there was no time-point observation and comparison.ALI model was established by intraperitoneal injection of lipopolysaccharide(...The model of acute lung injury(ALI)was established by intraperitoneal administration,but there was no time-point observation and comparison.ALI model was established by intraperitoneal injection of lipopolysaccharide(LPS)at the concentration of 10 mg·kg^-1 (10 mg LPS dissolved in 1 mL normal saline to prepare 1 mL·kg^-1solution)in rats.The control group(CG)was intraperitoneally injected with saline of the same dose.In the LPS group,lung tissues were collected at 4,6,8,12 and 24 h after administration.Then,the morphology changes,the ratio of wet-to-dry weight(W/D),the expression of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)proteins,the levels of malondialdehyde(MDA),the activities of superoxide dismutase(SOD),glutathione peroxidase(GSH)were measured.To verify the success of the model,the degrees of lung injury via Western blot,RT-PCR,ELISA and other techniques were detected at different time points,and the severe time of the ALI model established was deterimined by intraperitoneal administration,which provided a stable model basis for the study of the pathogenesis of ALI in the future.The results showed that the lung injury occurred in LPS group.W/D and lung pathological changes at 12 and 24 h of LPS group were significantly different from those in the CG.Compared with the CG,the expression of IL-1βand TNF-αproteins and the content of MDA in lung tissues of LPS group increased and most significant difference was found at 12 and 24 h(p<0.01).Compared with the CG,the activities of SOD and GSH in LPS 12 h group decreased significantly(p<0.01).In conclusion,inflammation and oxidative damage were the main causes of the ALI in rats.Lung injury was most obvious 12 h after intraperitoneal injection of 10 mg·kg^-1 LPS.展开更多
Objective To observe the therapeutic effects of intraperitoneal injection of antibiotics,intravenous injection of terlipressin,and combined treatment of coloclysis and plasma exchange on hepatic failure(HF),the subjec...Objective To observe the therapeutic effects of intraperitoneal injection of antibiotics,intravenous injection of terlipressin,and combined treatment of coloclysis and plasma exchange on hepatic failure(HF),the subjects included 494 inpatient cases of hepatic failure who were treated in Department of Infectious Diseases,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,China from 1997 to2008.Methods The patients that met the inclusion criteria were divided into intraperitoneal antibiotic injection group,intravenous terlipressin injection group,coloclysis group,plasma exchange group,combination group of coloclysis and plasma exchange in terms of treatment given and a control group was set up for each of the treatment group.In the intraperitoneal injection group,the prognosis and changes in clinical manifestations were observed in HF patients complicated with spontaneous peritonitis(SBP).In terlipressin injection group,HF patients complicated with hepatorenal syndrome(HRS) were observed for prognosis and changes in serum creatinine.In the combination group,the improvement in serum total bilirubin and prothrombin activity were observed.Results Two weeks after intraperitoneal injection of antibiotics,the ease ratios of abdominal pain,pressure pain and rebound tenderness were 87.64%,82.02%and 82.02%in the intraperitoneal injection group,respectively and the volume of ascites obviously decreased in 69 patients(77.53%).The survival rate in intraperitoneal injection group was significantly higher than in control group(P = 0.004).Four to eight days after the intravenous injection of terlipressin,the survival rate and the rate of serum creatinine decline of the treatment group were significantly higher than those in the control group(P = 0.003,P = 0.000).After 4 weeks of treatment,the ratio of clinical symptoms improvement(acratia,anorexia,abdominal distension,constipation) in coloclysis group were60.27%,57.53%,91.78%and 94.52%,in plasma exchange group were 71.83%,69.44%,75%and 72.22%,and in combination group were 82.14%,79.46%,92.85%and 95.54%.The serum total bilirubin was decreased and the prothrombin activity increased and the differences were statistically significant as compared with control group(P= 0.000).Conclusions The intraperitoneal injection of antibiotics,intravenous injection of terlipressin and combined treatment of coloclysis and plasma exchange were all effective for the treatment of HF and its complications.展开更多
Objective:Malignant peritoneal mesothelioma(MPM)is a rare primary malignant tumor with an extremely poor prognosis that currently lacks effective treatment options.This study investigated the in vitro and in vivo effi...Objective:Malignant peritoneal mesothelioma(MPM)is a rare primary malignant tumor with an extremely poor prognosis that currently lacks effective treatment options.This study investigated the in vitro and in vivo efficacy of natural killer(NK)cells for treatment of MPM.Methods:An in vitro study was conducted to assess the cytotoxicity of NK cells from umbilical cord blood to MPM cells with the use of a high-content imaging analysis system,the Cell Counting Kit-8 assay,and Wright-Giemsa staining.The level of NK cell effector molecule expression was detected by flow cytometry and enzyme-linked immunosorbent assays.The ability of NK cells to kill MPM cells was determined based on live cell imaging,transmission electron microscopy,and scanning electron microscopy.An in vivo study was conducted to assess the efficacy and safety of NK cell therapy based on the experimental peritoneal cancer index,small animal magnetic resonance imaging,and conventional histopathologic,cytologic,and hematologic studies.Results:NK cells effectively killed MPM cells through the release of effector molecules(granzyme B,perforin,interferon-γ,and tumor necrosis factor-α)in a dose-and density-dependent manner.The NK cell killing process potentially involved four dynamic steps:chemotaxis;hitting;adhesion;and penetration.NK cells significantly reduced the tumor burden,diminished ascites production,and extended survival with no significant hematologic toxicity or organ damage in NOG mice.Conclusions:NK cell immunotherapy inhibited proliferation of MPM cells in vitro and in vivo with a good safety profile.展开更多
AIM To compare the effects of intravenous routeand peritoneal route on liver targeted uptake andexpression of plasmid delivered by galactose-terminal glyco-poly-L-lysine(G-PLL).METHODS The plasmid pTM/MMP-1 which coul...AIM To compare the effects of intravenous routeand peritoneal route on liver targeted uptake andexpression of plasmid delivered by galactose-terminal glyco-poly-L-lysine(G-PLL).METHODS The plasmid pTM/MMP-1 which couldbe expressed in eukaryotic cells was bound to G-PLL,and was then transferred into Wistar rats byintravenous and intraperitoneal injection.Theexpression and distribution of the plasmid wereobserved at different time periods by in situhybridization and immunohistochemistry.RESULTS The plasmid could be expressedsignificantly within 24 h after being transferred invivo by both intravenous and intraperitonealroutes.One week later the expression began todecrease,and could still be observed three weekslater.Although both the intravenous andintraperitoneal route could target-specificallydeliver the plasmid to the liver,the effect of theformer was better as compared to that of the latter.CONCLUSION Intravenous route is better for livertargeted uptake and expression of G-PLL-boundplasmids than the peritoneal route.展开更多
Despite medical treatment,the lethality of severe acute pancreatitis is still high (20-30%).Therefore,it is very important to find good animal models to characterise the events of this severe disease.In 1984,Mizunuma ...Despite medical treatment,the lethality of severe acute pancreatitis is still high (20-30%).Therefore,it is very important to find good animal models to characterise the events of this severe disease.In 1984,Mizunuma et al. developed a new type of experimental necrotizing pancreatitis by intraperitoneal administration of a high dose of L-arginine in rats.This non-invasive model is highly reproducible and produces selective,dose-dependent acinar cell necrosis. Not only is this a good model to study the pathomechanisms of acute necrotizing pancreatitis,but it is also excellent to observe and influence the time course changes of the disease.By writing this review we iluminate some new aspects of cell physiology and pathology of acute necrotizing pancreatitis.Unfortunately,the reviews about acute experimental pancreatitis usually did not discuss this model. Therefore,the aim of this manuscript was to summarise the observations and address some challenges for the future in L-arginine-induced pancreatitis.展开更多
The authors declare that in the article1 published in vol.2,issue 4 of the Chinese Medical Journal Pulmonary and Critical Care Medicine,the description about“Before sacrifice,the mice were anesthetized with an intrap...The authors declare that in the article1 published in vol.2,issue 4 of the Chinese Medical Journal Pulmonary and Critical Care Medicine,the description about“Before sacrifice,the mice were anesthetized with an intraperitoneal injection(i.p.)injection of 4%chloral hydrate at a dose of 0.1 mL per 10 g of body weight.”presented in the“Transgenic mouse and lung injury models”section on Page 266 should be corrected to“Before sacrifice,the mice were anesthetized with an intraperitoneal injection(i.p.)of 0.3%pentobarbital sodium at a dose of 50 mg/kg”.The authors deeply apologize for any inconvenience caused.展开更多
Objective To investigate the underlying mechanism of“Zhibian”(BL54)-toward-"Shuidao"(ST28)acupuncturettechniqueforimproving reproductive function in mice with asthenospermia by regulating ferroptosis pathw...Objective To investigate the underlying mechanism of“Zhibian”(BL54)-toward-"Shuidao"(ST28)acupuncturettechniqueforimproving reproductive function in mice with asthenospermia by regulating ferroptosis pathway.Methods Sixty male C57BL/6 mice were randomly divided into a blank group,a model group,an acupuncture group and a Fer-1 group,15 mice in each one.Except the blank group,the intraperitoneal injection with cyclophosphamide(50·kg-1.d-')was administered to establish the asthenospermia model.in the mice of the rest 3 groups for 5 consecutive days.In the acupuncture group,"Zhibian"(BL54)-toward-"Shuidao"(ST28)acupuncture technique was operated in the mice,for 20 min each time;and in the Fer-l group,Fer-1 solution(1 mg/kg)was injected intraperitoneally.The interventions of these two groups were delivered once daily and for 2 consecutive weeks.The testicular wet weight was measured and the testicular coefficient was calculated.Using sperm quality detection system,the sperm quality was detected.With ELISA used,the contents of testosterone(T),follicle-stimulating hormone(FSH)and luteinizing hormone(LH)in the serum were detected.With HE staining,testicular and epididymal morphology was observed.Immunofluorescence was used to detect the expression of reactive oxygen species(ROS)in the testes.Biochemical assay was conducted to determine the contents of malondialdehyde(MDA),reduced glutathione(GSH),and total iron ion(TFe)in the testicular tissue.Transmission electron microscopy was used to examine mitochondrial structure of the testis,while JC-1 staining was used to assess mitochondrial membrane potential in the testicular tissue.Fluorescence quantitative PCR and Western blot analyseswere employed to measure the mRNA and protein expression of solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),ferritin heavy chain 1(FTH1),and acyl-CoA synthetase long-chain family member 4(ACSL4)in the testicular tissue.Results Compared with the blank group,in the model group,the testicular wet weight and testicular coefficient decreased(P<0.01);the sperm concentration and sperm motility reduced(P<0.01),and the contents of T,FSH,and LH decreased in the serum(P<0.01);and the seminiferous tubules in the testis showed loose structure and deformed lumen,sperm cells were disorganized and the sperm numbers reduced;the tubular walls became thinner,and sperm numbers in the lumen less;the expression of ROS in testicular tissue,as well as the contents of MDA and TFe increased(P<0.01),and the content of GSH decreased(P<0.01);and the numbers of mitochondria reduced,the structure of cristae was serious damaged,and mitochondrial membrane potential level declined(P<0.O1);the mRNA and protein expression of SLC7A11,GPX4,and FTH1 decreased(P<0.01),while the mRNA and protein expression of ACSL4 increased(P<0.01).In comparison with the model group,the acupuncture and Fer-1 groups showed the increase of testicular wet weight and coefficient(P<0.01),sperm concentration and motility(P<0.01),and the serum contents of T,FSH,and LH(P<0.01);and the improvements in testicular and epididymal histopathology;ROS expression and the contents of MDA and TFe decreased(P<0.01),and the content of GSH elevated(P<0.05);the mitochondrial structure and numbers were ameliorated and mitochondrial membrane potential rose(P<0.01).Besides,in comparison with the model group,the mRNA expression of SLC7A1l was higher(P<0.05,P<0.01),the mRNA and protein expression of GPX4 and FTH1 increased(P<0.01,P<0.05),and the mRNA and protein expression of ACSL4 decreased(P<0.01)in the acupuncture and the Fer-1 groups;and the protein expression of SLC7A11 was higher in the Fer-1 group(P<0.05).Conclusion"Zhibian"(BL54)-toward-"Shuidao"(ST28)acupuncture technique may improve the reproductive capacity in the mice with asthenospermia by alleviating ferroptosis-induced cellular damage and ameliorating testicular function.展开更多
文摘Wen-lin Gong1,Chuang Sha2,Gang Du1,Zhong-gui Shan3,Zhong-quan Qi3,Su-fang Zhou1,Nuo Yang1,4,Yong-xiang Zhao1,4.First published:21 June 2017;10(5):454-460.DOI:10.1016/j.apjtm.2017.05.004 The authors would like to correct an error in Figure 3 in which the flow cytometric scattergram of CD4/CD44 for the control group was erroneously used for the scattergram of CD8/CD44 for the PVIDSC group.The correct scattergram of CD8/CD44 for the PVIDSC group is provided below.The error does not affect the conclusion of the study.The authors apologize for the error and the inconvenience it might have caused to readers.
基金Supported by National High-Tech Research and Development Program of China (863 Program, No.2001AA5070, 2002AA639600)Natural Science Foundation of Fujian Province of China (No.B0410022, 2006F5066)
文摘Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups.Each bullfrog in bacterium-injected group was injected intraperitoneally(i.p.) with 0.2 ml bacterial suspension at a density of 5.2 × 106 CFU/ml,while each one in control group injected i.p.with 0.2 ml sterile saline solution(0.85%,w/v).Three bullfrogs in both groups were sampled at 0,1,3,7,11,15 and 20 days post-injection(dpi) for the evaluation of non-specific immune parameters.It was observed that intraperitoneal injection of A.hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood.Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group.However,a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes.No significant difference in changes in the number of peripheral erythrocytes,serum superoxide dismutase(SOD) activity,and lysozyme activity was detected between the two groups.It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A.hydrophila infection.
基金Supported by the National Key Research and Development Program of China(2016YED0501008)the National Natural Science Foundation of China(31772806)the Natural Science Foundation of Heilongjiang Province(C2017022)。
文摘The model of acute lung injury(ALI)was established by intraperitoneal administration,but there was no time-point observation and comparison.ALI model was established by intraperitoneal injection of lipopolysaccharide(LPS)at the concentration of 10 mg·kg^-1 (10 mg LPS dissolved in 1 mL normal saline to prepare 1 mL·kg^-1solution)in rats.The control group(CG)was intraperitoneally injected with saline of the same dose.In the LPS group,lung tissues were collected at 4,6,8,12 and 24 h after administration.Then,the morphology changes,the ratio of wet-to-dry weight(W/D),the expression of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)proteins,the levels of malondialdehyde(MDA),the activities of superoxide dismutase(SOD),glutathione peroxidase(GSH)were measured.To verify the success of the model,the degrees of lung injury via Western blot,RT-PCR,ELISA and other techniques were detected at different time points,and the severe time of the ALI model established was deterimined by intraperitoneal administration,which provided a stable model basis for the study of the pathogenesis of ALI in the future.The results showed that the lung injury occurred in LPS group.W/D and lung pathological changes at 12 and 24 h of LPS group were significantly different from those in the CG.Compared with the CG,the expression of IL-1βand TNF-αproteins and the content of MDA in lung tissues of LPS group increased and most significant difference was found at 12 and 24 h(p<0.01).Compared with the CG,the activities of SOD and GSH in LPS 12 h group decreased significantly(p<0.01).In conclusion,inflammation and oxidative damage were the main causes of the ALI in rats.Lung injury was most obvious 12 h after intraperitoneal injection of 10 mg·kg^-1 LPS.
文摘Objective To observe the therapeutic effects of intraperitoneal injection of antibiotics,intravenous injection of terlipressin,and combined treatment of coloclysis and plasma exchange on hepatic failure(HF),the subjects included 494 inpatient cases of hepatic failure who were treated in Department of Infectious Diseases,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,China from 1997 to2008.Methods The patients that met the inclusion criteria were divided into intraperitoneal antibiotic injection group,intravenous terlipressin injection group,coloclysis group,plasma exchange group,combination group of coloclysis and plasma exchange in terms of treatment given and a control group was set up for each of the treatment group.In the intraperitoneal injection group,the prognosis and changes in clinical manifestations were observed in HF patients complicated with spontaneous peritonitis(SBP).In terlipressin injection group,HF patients complicated with hepatorenal syndrome(HRS) were observed for prognosis and changes in serum creatinine.In the combination group,the improvement in serum total bilirubin and prothrombin activity were observed.Results Two weeks after intraperitoneal injection of antibiotics,the ease ratios of abdominal pain,pressure pain and rebound tenderness were 87.64%,82.02%and 82.02%in the intraperitoneal injection group,respectively and the volume of ascites obviously decreased in 69 patients(77.53%).The survival rate in intraperitoneal injection group was significantly higher than in control group(P = 0.004).Four to eight days after the intravenous injection of terlipressin,the survival rate and the rate of serum creatinine decline of the treatment group were significantly higher than those in the control group(P = 0.003,P = 0.000).After 4 weeks of treatment,the ratio of clinical symptoms improvement(acratia,anorexia,abdominal distension,constipation) in coloclysis group were60.27%,57.53%,91.78%and 94.52%,in plasma exchange group were 71.83%,69.44%,75%and 72.22%,and in combination group were 82.14%,79.46%,92.85%and 95.54%.The serum total bilirubin was decreased and the prothrombin activity increased and the differences were statistically significant as compared with control group(P= 0.000).Conclusions The intraperitoneal injection of antibiotics,intravenous injection of terlipressin and combined treatment of coloclysis and plasma exchange were all effective for the treatment of HF and its complications.
基金supported by the General Program of National Natural Science Foundation of China(Grant No.82073376)。
文摘Objective:Malignant peritoneal mesothelioma(MPM)is a rare primary malignant tumor with an extremely poor prognosis that currently lacks effective treatment options.This study investigated the in vitro and in vivo efficacy of natural killer(NK)cells for treatment of MPM.Methods:An in vitro study was conducted to assess the cytotoxicity of NK cells from umbilical cord blood to MPM cells with the use of a high-content imaging analysis system,the Cell Counting Kit-8 assay,and Wright-Giemsa staining.The level of NK cell effector molecule expression was detected by flow cytometry and enzyme-linked immunosorbent assays.The ability of NK cells to kill MPM cells was determined based on live cell imaging,transmission electron microscopy,and scanning electron microscopy.An in vivo study was conducted to assess the efficacy and safety of NK cell therapy based on the experimental peritoneal cancer index,small animal magnetic resonance imaging,and conventional histopathologic,cytologic,and hematologic studies.Results:NK cells effectively killed MPM cells through the release of effector molecules(granzyme B,perforin,interferon-γ,and tumor necrosis factor-α)in a dose-and density-dependent manner.The NK cell killing process potentially involved four dynamic steps:chemotaxis;hitting;adhesion;and penetration.NK cells significantly reduced the tumor burden,diminished ascites production,and extended survival with no significant hematologic toxicity or organ damage in NOG mice.Conclusions:NK cell immunotherapy inhibited proliferation of MPM cells in vitro and in vivo with a good safety profile.
基金National Natural Science Foundation of China(№39570336).
文摘AIM To compare the effects of intravenous routeand peritoneal route on liver targeted uptake andexpression of plasmid delivered by galactose-terminal glyco-poly-L-lysine(G-PLL).METHODS The plasmid pTM/MMP-1 which couldbe expressed in eukaryotic cells was bound to G-PLL,and was then transferred into Wistar rats byintravenous and intraperitoneal injection.Theexpression and distribution of the plasmid wereobserved at different time periods by in situhybridization and immunohistochemistry.RESULTS The plasmid could be expressedsignificantly within 24 h after being transferred invivo by both intravenous and intraperitonealroutes.One week later the expression began todecrease,and could still be observed three weekslater.Although both the intravenous andintraperitoneal route could target-specificallydeliver the plasmid to the liver,the effect of theformer was better as compared to that of the latter.CONCLUSION Intravenous route is better for livertargeted uptake and expression of G-PLL-boundplasmids than the peritoneal route.
基金Supported by The Wellcome Trust,Grant No.022618,and by the Hungarian Scientific Research Fund,No.D42188,T43066 and T042589
文摘Despite medical treatment,the lethality of severe acute pancreatitis is still high (20-30%).Therefore,it is very important to find good animal models to characterise the events of this severe disease.In 1984,Mizunuma et al. developed a new type of experimental necrotizing pancreatitis by intraperitoneal administration of a high dose of L-arginine in rats.This non-invasive model is highly reproducible and produces selective,dose-dependent acinar cell necrosis. Not only is this a good model to study the pathomechanisms of acute necrotizing pancreatitis,but it is also excellent to observe and influence the time course changes of the disease.By writing this review we iluminate some new aspects of cell physiology and pathology of acute necrotizing pancreatitis.Unfortunately,the reviews about acute experimental pancreatitis usually did not discuss this model. Therefore,the aim of this manuscript was to summarise the observations and address some challenges for the future in L-arginine-induced pancreatitis.
文摘The authors declare that in the article1 published in vol.2,issue 4 of the Chinese Medical Journal Pulmonary and Critical Care Medicine,the description about“Before sacrifice,the mice were anesthetized with an intraperitoneal injection(i.p.)injection of 4%chloral hydrate at a dose of 0.1 mL per 10 g of body weight.”presented in the“Transgenic mouse and lung injury models”section on Page 266 should be corrected to“Before sacrifice,the mice were anesthetized with an intraperitoneal injection(i.p.)of 0.3%pentobarbital sodium at a dose of 50 mg/kg”.The authors deeply apologize for any inconvenience caused.
文摘Objective To investigate the underlying mechanism of“Zhibian”(BL54)-toward-"Shuidao"(ST28)acupuncturettechniqueforimproving reproductive function in mice with asthenospermia by regulating ferroptosis pathway.Methods Sixty male C57BL/6 mice were randomly divided into a blank group,a model group,an acupuncture group and a Fer-1 group,15 mice in each one.Except the blank group,the intraperitoneal injection with cyclophosphamide(50·kg-1.d-')was administered to establish the asthenospermia model.in the mice of the rest 3 groups for 5 consecutive days.In the acupuncture group,"Zhibian"(BL54)-toward-"Shuidao"(ST28)acupuncture technique was operated in the mice,for 20 min each time;and in the Fer-l group,Fer-1 solution(1 mg/kg)was injected intraperitoneally.The interventions of these two groups were delivered once daily and for 2 consecutive weeks.The testicular wet weight was measured and the testicular coefficient was calculated.Using sperm quality detection system,the sperm quality was detected.With ELISA used,the contents of testosterone(T),follicle-stimulating hormone(FSH)and luteinizing hormone(LH)in the serum were detected.With HE staining,testicular and epididymal morphology was observed.Immunofluorescence was used to detect the expression of reactive oxygen species(ROS)in the testes.Biochemical assay was conducted to determine the contents of malondialdehyde(MDA),reduced glutathione(GSH),and total iron ion(TFe)in the testicular tissue.Transmission electron microscopy was used to examine mitochondrial structure of the testis,while JC-1 staining was used to assess mitochondrial membrane potential in the testicular tissue.Fluorescence quantitative PCR and Western blot analyseswere employed to measure the mRNA and protein expression of solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),ferritin heavy chain 1(FTH1),and acyl-CoA synthetase long-chain family member 4(ACSL4)in the testicular tissue.Results Compared with the blank group,in the model group,the testicular wet weight and testicular coefficient decreased(P<0.01);the sperm concentration and sperm motility reduced(P<0.01),and the contents of T,FSH,and LH decreased in the serum(P<0.01);and the seminiferous tubules in the testis showed loose structure and deformed lumen,sperm cells were disorganized and the sperm numbers reduced;the tubular walls became thinner,and sperm numbers in the lumen less;the expression of ROS in testicular tissue,as well as the contents of MDA and TFe increased(P<0.01),and the content of GSH decreased(P<0.01);and the numbers of mitochondria reduced,the structure of cristae was serious damaged,and mitochondrial membrane potential level declined(P<0.O1);the mRNA and protein expression of SLC7A11,GPX4,and FTH1 decreased(P<0.01),while the mRNA and protein expression of ACSL4 increased(P<0.01).In comparison with the model group,the acupuncture and Fer-1 groups showed the increase of testicular wet weight and coefficient(P<0.01),sperm concentration and motility(P<0.01),and the serum contents of T,FSH,and LH(P<0.01);and the improvements in testicular and epididymal histopathology;ROS expression and the contents of MDA and TFe decreased(P<0.01),and the content of GSH elevated(P<0.05);the mitochondrial structure and numbers were ameliorated and mitochondrial membrane potential rose(P<0.01).Besides,in comparison with the model group,the mRNA expression of SLC7A1l was higher(P<0.05,P<0.01),the mRNA and protein expression of GPX4 and FTH1 increased(P<0.01,P<0.05),and the mRNA and protein expression of ACSL4 decreased(P<0.01)in the acupuncture and the Fer-1 groups;and the protein expression of SLC7A11 was higher in the Fer-1 group(P<0.05).Conclusion"Zhibian"(BL54)-toward-"Shuidao"(ST28)acupuncture technique may improve the reproductive capacity in the mice with asthenospermia by alleviating ferroptosis-induced cellular damage and ameliorating testicular function.
