Objective:To explore the role of NLRP3/Caspase-1-mediated pyroptosis on peripheral blood monocytes in patients with essential hypertension(EH),and its regulation by pannexin-1(Panx-1)hemi-channels.Methods:The peripher...Objective:To explore the role of NLRP3/Caspase-1-mediated pyroptosis on peripheral blood monocytes in patients with essential hypertension(EH),and its regulation by pannexin-1(Panx-1)hemi-channels.Methods:The peripheral blood of EH patients and healthy subjects was collected,peripheral plasma of the two groups was subsequently separated,and IL-1βcontent in peripheral plasma was further measured by ELISA.In the meantime,the monocytes from peripheral blood were sorted by immunomagnetic beads,and mRNA and protein expression of Panx-1,NLRP3 inflammasome related molecules(NLRP3,ASC,Caspase-1),downstream effector IL-1β,and pyroptosis-related protein GSDMD,were analyzed by RT-qPCR and Western blot,respectively.Subsequently,human primary monocytes in two group were cultured in vitro.Immunofluorescence assay was performed to detect the expression and location of Panx-1 on monocytes.Finally,the cultured monocytes from EH patients were exposed to NLRP3 inflammasome activator lipopolysaccharide(LPS),and also to LPS pretreated with Panx-1 hemi-channel inhibitor probenecid or specific Panx-1 siRNA,followed by collection of the cell culture supernatant and monocytes.The cell viability of each group was detected by CCK-8 assay,IL-1βcontent in the culture supernatant was analyzed by ELISA,and the expression of target proteins on monocytes was determined by Western blot assay.Results:Compared with healthy subjects,EH patients showed higher IL-1βcontent in peripheral plasma,and increased expression in both mRNA and protein levels for Panx-1,NLRP3 inflammasome related molecules,IL-1βand GSDMD on peripheral monocytes.Furthermore,Panx-1 protein in EH patients was significantly higher than that in healthy subjects and localized on monocyte membrane as evidenced by immunofluorescence assay.In the cultured human primary monocytes from EH patients,the cell viability in each group showed no significant difference.LPS stimulation can induce NLRP3-dependent pyroptosis,by showing an increase in IL-1βcontent in cell culture supernatant and protein expressions of NLRP3 inflammasome related molecules and GSDMD on monocytes,by LPS exposure.However,the above stimulating effect of LPS could be counteracted by pretreatment with probenecid or Panx-1 siRNA.Conclusion:The NLRP3/Caspase-1-mediated pyroptosis is activated on peripheral blood monocytes in patients with EH,and this activation could be regulated by Panx-1 hemi-channels.展开更多
Objective: To determine serum pannexin-1 channel levels and their association with hair loss in women with PCOS diagnosed with female androgenetic alopecia (FAGA). Materials and Methods: Thirty-five women with PCOS wh...Objective: To determine serum pannexin-1 channel levels and their association with hair loss in women with PCOS diagnosed with female androgenetic alopecia (FAGA). Materials and Methods: Thirty-five women with PCOS who presented with diffuse and treatment-resistant progressive hair loss and were diagnosed with FAGA were included in the study. 25 patients who were diagnosed with female androgenetic alopecia but did not have PCOS were considered as the control group. PCOS and control groups were matched by age. Follicular miniaturization, displacement of terminal hairs with vellus hairs, and a diffuse decrease in hair density were accepted as FAGA in the trcihoscopy examination of the vertex and bitempoaral area. On the third day of the menstrual cycle serum FSH, LH, testosterone, PRL and insulin levels were measured. Insulin resistance was calculated with HOMA-IR. Serum pannexin-1 channel levels of each group were mesured with ELISA. Results: Serum pannexin 1 channels levels of FAGA group due to PCOS were found to be significantly higher than FAGA patients in the control group (2.72 ± 1.09 ng/mL vs 1.65 ± 0.97 ng/mL, p < 0.01). Serum LH, insulin and testosterone levels of PCOS group were significantly higher than controls. HOMA-IR values were significantly higher and >2.5 in the PCOS group compared to the controls. PRL values were similar except for one patient with elevated PRL. Serum FSH values were the same in both groups. A positive and significant correlation was found between pannexin 1 channels levels and HOMA-IR and serum testosterone levels (r = 0.650, p Conclusions: In addition to hyperandrogenemia, increased pannexin 1 channel levels may play a role in the etiology of PCOS associated FAGA, as it impairs the communication between the skin and hair follicle.展开更多
背景:程序性细胞死亡受体1(programmed death receptor-1,PD-1)在高糖环境下影响骨髓间充质干细胞成骨分化的作用机制尚不清楚。目的:探讨高糖环境中PD-1对大鼠骨髓间充质干细胞成骨分化的影响及其调控机制。方法:将大鼠骨髓间充质干细...背景:程序性细胞死亡受体1(programmed death receptor-1,PD-1)在高糖环境下影响骨髓间充质干细胞成骨分化的作用机制尚不清楚。目的:探讨高糖环境中PD-1对大鼠骨髓间充质干细胞成骨分化的影响及其调控机制。方法:将大鼠骨髓间充质干细胞随机分为正常糖组(5.6 mmol/L)、高糖组(30 mmol/L)、PD-1过表达组、PD-1过表达空载组、PD-1敲低组、PD-1敲低空载组、PI3K/AKT通路抑制剂组(PD-1敲低+5μmol/L LY294002)。通过在高糖培养基中培养大鼠骨髓间充质干细胞来模拟体外糖尿病环境,采用qRT-PCR检测大鼠骨髓间充质干细胞中PD-1及其配体PD-L1和成骨标志物Runx2、OSX的mRNA表达,采用碱性磷酸酶染色和茜素红S染色观察成骨分化能力,采用CCK-8检测细胞增殖情况,采用Western blot检测PD-1、PD-L1、p-PI3K、p-AKT的蛋白表达。结果与结论:①高糖组PD-1及PD-L1表达显著高于正常糖组,高糖组骨髓间充质干细胞的成骨分化能力较正常糖组显著下降;②敲低PD-1表达可以促进骨髓间充质干细胞的成骨分化、增加细胞增殖活性,同时激活PI3K/AKT通路;③加入PI3K/AKT通路抑制剂LY294002后,骨髓间充质干细胞成骨分化能力显著下降。结果表明:PD-1依赖于PI3K/AKT信号通路抑制高糖环境下大鼠骨髓间充质干细胞的成骨分化。展开更多
基金National Natural Science Foundation of China(No.81970313)Natural Science Foundation of Anhui Province(No.2008085MH238)+1 种基金512 Talent Cultivation Plan of Bengbu Medical College(No.by51201321)Key Program of Natural Science Foundation of Bengbu Medical College(No.2020byzd109)。
文摘Objective:To explore the role of NLRP3/Caspase-1-mediated pyroptosis on peripheral blood monocytes in patients with essential hypertension(EH),and its regulation by pannexin-1(Panx-1)hemi-channels.Methods:The peripheral blood of EH patients and healthy subjects was collected,peripheral plasma of the two groups was subsequently separated,and IL-1βcontent in peripheral plasma was further measured by ELISA.In the meantime,the monocytes from peripheral blood were sorted by immunomagnetic beads,and mRNA and protein expression of Panx-1,NLRP3 inflammasome related molecules(NLRP3,ASC,Caspase-1),downstream effector IL-1β,and pyroptosis-related protein GSDMD,were analyzed by RT-qPCR and Western blot,respectively.Subsequently,human primary monocytes in two group were cultured in vitro.Immunofluorescence assay was performed to detect the expression and location of Panx-1 on monocytes.Finally,the cultured monocytes from EH patients were exposed to NLRP3 inflammasome activator lipopolysaccharide(LPS),and also to LPS pretreated with Panx-1 hemi-channel inhibitor probenecid or specific Panx-1 siRNA,followed by collection of the cell culture supernatant and monocytes.The cell viability of each group was detected by CCK-8 assay,IL-1βcontent in the culture supernatant was analyzed by ELISA,and the expression of target proteins on monocytes was determined by Western blot assay.Results:Compared with healthy subjects,EH patients showed higher IL-1βcontent in peripheral plasma,and increased expression in both mRNA and protein levels for Panx-1,NLRP3 inflammasome related molecules,IL-1βand GSDMD on peripheral monocytes.Furthermore,Panx-1 protein in EH patients was significantly higher than that in healthy subjects and localized on monocyte membrane as evidenced by immunofluorescence assay.In the cultured human primary monocytes from EH patients,the cell viability in each group showed no significant difference.LPS stimulation can induce NLRP3-dependent pyroptosis,by showing an increase in IL-1βcontent in cell culture supernatant and protein expressions of NLRP3 inflammasome related molecules and GSDMD on monocytes,by LPS exposure.However,the above stimulating effect of LPS could be counteracted by pretreatment with probenecid or Panx-1 siRNA.Conclusion:The NLRP3/Caspase-1-mediated pyroptosis is activated on peripheral blood monocytes in patients with EH,and this activation could be regulated by Panx-1 hemi-channels.
文摘Objective: To determine serum pannexin-1 channel levels and their association with hair loss in women with PCOS diagnosed with female androgenetic alopecia (FAGA). Materials and Methods: Thirty-five women with PCOS who presented with diffuse and treatment-resistant progressive hair loss and were diagnosed with FAGA were included in the study. 25 patients who were diagnosed with female androgenetic alopecia but did not have PCOS were considered as the control group. PCOS and control groups were matched by age. Follicular miniaturization, displacement of terminal hairs with vellus hairs, and a diffuse decrease in hair density were accepted as FAGA in the trcihoscopy examination of the vertex and bitempoaral area. On the third day of the menstrual cycle serum FSH, LH, testosterone, PRL and insulin levels were measured. Insulin resistance was calculated with HOMA-IR. Serum pannexin-1 channel levels of each group were mesured with ELISA. Results: Serum pannexin 1 channels levels of FAGA group due to PCOS were found to be significantly higher than FAGA patients in the control group (2.72 ± 1.09 ng/mL vs 1.65 ± 0.97 ng/mL, p < 0.01). Serum LH, insulin and testosterone levels of PCOS group were significantly higher than controls. HOMA-IR values were significantly higher and >2.5 in the PCOS group compared to the controls. PRL values were similar except for one patient with elevated PRL. Serum FSH values were the same in both groups. A positive and significant correlation was found between pannexin 1 channels levels and HOMA-IR and serum testosterone levels (r = 0.650, p Conclusions: In addition to hyperandrogenemia, increased pannexin 1 channel levels may play a role in the etiology of PCOS associated FAGA, as it impairs the communication between the skin and hair follicle.
