Objective To evaluate the effect of Guilu Erxian Glue(龟鹿二仙胶,GEG)on cyclophosphamide(CTX)-induced bone marrow hematopoietic stem cells(HSCs)senescence in mice and explore the underlying mechanism.Methods The H22 l...Objective To evaluate the effect of Guilu Erxian Glue(龟鹿二仙胶,GEG)on cyclophosphamide(CTX)-induced bone marrow hematopoietic stem cells(HSCs)senescence in mice and explore the underlying mechanism.Methods The H22 liver cancer ascites lump model was established in male Kunming mice by injecting intraperitoneally(i.p.)with 5×10^6/mL H22 cells per mouse.Fifty tumor-bearing mice were divided into the control,model,pifithrin-α,GEG,and GEG+pifithrin-αgroups using a random number table,10 mice in each group.CTX(100 mg/kg i.p.)was administrated to mice from day 1 to day 3(d1–d3)continuously except for the control group.The mice in the pifithrin-α,GEG and GEG+pifithrin-αgroups were treated with pifithrin-α(2.2 mg/(kg·d)i.p.)for 6 consecutive days(d4–d9),GEG(9.5 g/(kg·d)i.p.)for 9 consecutive days(d1–d9),and GEG plus pifithrin-α,respectively.HSCs were collected after 9-d drug treatment.The anti-aging effect of GEG was studied by cell viability,cell cycle,andβ-galactosidase(β-gal)assays.The mRNA and protein expressions of cyclin-dependent kinase 2(CDK2),CDK4,inhibitor of cyclin-dependent kinase 4a encoding the tumor suppressor protein p16^(p16^INK4a),p21^Cip1/Waf1,p53,and phosphorylated retinoblastoma(pRb)were evaluated by quantitative real-time reverse transcription-polymerase chain reaction and semi-quantitative Western blot,respectively.Results Compared with the model group,GEG increased cell viability as well as proliferation(P<0.05 or P<0.01)and reducedβ-gal expression.Furthermore,GEG significantly decreased the expressions of p16^INK4a,p53 and p21^Cip1/Waf1 proteins,and increased the expressions of CDK2,CDK4 and pRb proteins compared with the model group(P<0.05 or P<0.01).Conclusion GEG can alleviate CTX-induced HSCs senescence in mice,and the p16^INK4a-Rb signaling pathway might be the underlying mechanism.展开更多
目的:通过检测慢性宫颈炎(chronic cervicitis)、宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)及宫颈鳞癌(squamous carcinoma of the cervix,SCC)组织中p16INK4A蛋白表达与人乳头瘤病毒(human papillomavirus,HPV)感染的状...目的:通过检测慢性宫颈炎(chronic cervicitis)、宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)及宫颈鳞癌(squamous carcinoma of the cervix,SCC)组织中p16INK4A蛋白表达与人乳头瘤病毒(human papillomavirus,HPV)感染的状况,探讨p16INK4A蛋白和HPV感染与宫颈鳞癌发生发展的关系。方法:采用免疫组化PV-9000方法检测114例宫颈组织标本的p16INK4A表达,用核酸分子快速导流杂交基因芯片技术(HybriMax)检测21种HPV DNA。结果:(1)慢性宫颈炎、CINⅠ、CINⅡ、CINⅢ、SCC组织中p16INK4A蛋白表达阳性率分别为0%、45.83%、72.73%、84.00%、91.30%,随着宫颈病变程度加重,p16INK4A蛋白表达阳性率逐渐增高,表达强度增加,差异有统计学意义(P<0.05)。(2)慢性宫颈炎、CINI、CINⅡ、CINⅢ、SCC中HPV感染阳性率分别为20.00%、37.50%、54.55%、56.00%、73.91%,HPV感染在不同宫颈病变组织中差异有统计学意义(P<0.05)。随宫颈病变程度加重,HPV阳性率呈递增趋势,不同病变组织中位于前3位的HPV感染型别分别为:CINⅠ组,HPV16、18、58;CINⅡ～Ⅲ组,HPV16、33、52;宫颈鳞癌组,HPV16、18,52。(3)相关分析结果显示,病变组织中p16INK4A蛋白表达与HPV感染呈正相关(r=0.268,P<0.05)。结论:p16INK4A蛋白表达与宫颈病变程度有关,p16INK4A可能参与了HPV相关的宫颈癌发生。二者联合检测对宫颈癌筛查和预防具有重要意义。展开更多
基金Supported by the National Natural Science Foundation of 6hina(No.81904197)Natural Science Foundation of Zhejiang Province(No.LQ15H290002)and 2019 Research and Innovation Fund Project for Young and Middle-aged Researchers of Zhejiang Chinese Medical University(No.KC201944)。
文摘Objective To evaluate the effect of Guilu Erxian Glue(龟鹿二仙胶,GEG)on cyclophosphamide(CTX)-induced bone marrow hematopoietic stem cells(HSCs)senescence in mice and explore the underlying mechanism.Methods The H22 liver cancer ascites lump model was established in male Kunming mice by injecting intraperitoneally(i.p.)with 5×10^6/mL H22 cells per mouse.Fifty tumor-bearing mice were divided into the control,model,pifithrin-α,GEG,and GEG+pifithrin-αgroups using a random number table,10 mice in each group.CTX(100 mg/kg i.p.)was administrated to mice from day 1 to day 3(d1–d3)continuously except for the control group.The mice in the pifithrin-α,GEG and GEG+pifithrin-αgroups were treated with pifithrin-α(2.2 mg/(kg·d)i.p.)for 6 consecutive days(d4–d9),GEG(9.5 g/(kg·d)i.p.)for 9 consecutive days(d1–d9),and GEG plus pifithrin-α,respectively.HSCs were collected after 9-d drug treatment.The anti-aging effect of GEG was studied by cell viability,cell cycle,andβ-galactosidase(β-gal)assays.The mRNA and protein expressions of cyclin-dependent kinase 2(CDK2),CDK4,inhibitor of cyclin-dependent kinase 4a encoding the tumor suppressor protein p16^(p16^INK4a),p21^Cip1/Waf1,p53,and phosphorylated retinoblastoma(pRb)were evaluated by quantitative real-time reverse transcription-polymerase chain reaction and semi-quantitative Western blot,respectively.Results Compared with the model group,GEG increased cell viability as well as proliferation(P<0.05 or P<0.01)and reducedβ-gal expression.Furthermore,GEG significantly decreased the expressions of p16^INK4a,p53 and p21^Cip1/Waf1 proteins,and increased the expressions of CDK2,CDK4 and pRb proteins compared with the model group(P<0.05 or P<0.01).Conclusion GEG can alleviate CTX-induced HSCs senescence in mice,and the p16^INK4a-Rb signaling pathway might be the underlying mechanism.
文摘目的:通过检测慢性宫颈炎(chronic cervicitis)、宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)及宫颈鳞癌(squamous carcinoma of the cervix,SCC)组织中p16INK4A蛋白表达与人乳头瘤病毒(human papillomavirus,HPV)感染的状况,探讨p16INK4A蛋白和HPV感染与宫颈鳞癌发生发展的关系。方法:采用免疫组化PV-9000方法检测114例宫颈组织标本的p16INK4A表达,用核酸分子快速导流杂交基因芯片技术(HybriMax)检测21种HPV DNA。结果:(1)慢性宫颈炎、CINⅠ、CINⅡ、CINⅢ、SCC组织中p16INK4A蛋白表达阳性率分别为0%、45.83%、72.73%、84.00%、91.30%,随着宫颈病变程度加重,p16INK4A蛋白表达阳性率逐渐增高,表达强度增加,差异有统计学意义(P<0.05)。(2)慢性宫颈炎、CINI、CINⅡ、CINⅢ、SCC中HPV感染阳性率分别为20.00%、37.50%、54.55%、56.00%、73.91%,HPV感染在不同宫颈病变组织中差异有统计学意义(P<0.05)。随宫颈病变程度加重,HPV阳性率呈递增趋势,不同病变组织中位于前3位的HPV感染型别分别为:CINⅠ组,HPV16、18、58;CINⅡ～Ⅲ组,HPV16、33、52;宫颈鳞癌组,HPV16、18,52。(3)相关分析结果显示,病变组织中p16INK4A蛋白表达与HPV感染呈正相关(r=0.268,P<0.05)。结论:p16INK4A蛋白表达与宫颈病变程度有关,p16INK4A可能参与了HPV相关的宫颈癌发生。二者联合检测对宫颈癌筛查和预防具有重要意义。
