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Objective To study the role of endoplasmic reticulum stress in phenotypic change of cultured human glomerular mesangial cells induced by high glucose.Methods Cultured human glomeruar mesangial cells were divided into three groups: control group,high glucose group and high glucose+ 4-phenylbutyric acid (4-PBA) group.Cell number of proliferation was assessed by MTT assay.Cell cycle was measured by flow cytometric analysis.Expression of α-SMA was assessed by immunohistochemistry and was observed by laser scanning confocal microscope.Involved mRNA and protein expression were measured by real-time PCR and Western blotting.Results (1)Cell number of proliferation and S transition proportion in high glucose group significantly increased than that in control group (P ＜ 0.05).High glucose could induce α-SMA expression significantly (P＜0.05).4-PBA could significantly inhibit human glomerular mesangial cells proliferation (P＜0.05),S transition arrest (P＜0.05) and expression of α-SMA (P＜0.05) induced by high glucose.(2) Compared with control group,high glucose could significantly increase the expression of glucose-regulated protein78(Grp78 ) mRNA and protein (P＜ 0.05),which could be inhibited by 4-PBA treatment (P＜0.05).(3)High glucose could induce the mRNA and protein expression of TGF-β1 and FN significantly,which could be inhibited by 4-PBA treatment (P＜0.05).Conclusion Endoplasmic reticulum stress plays an important role in phenotypic change of cultured human glomerular mesangial cells induced by high glucose.
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Objective To investigate the expression of snrvivin,an anti-apoptosis gene,in breast cancer tissues and its relationship with clinical parameters of breast cancer patients.To investigate the role of survivin in oncogenesis of breast cancer and its relation with prognosis of breast cancer.Methods Western blot was conducted to detect survivin expression in cancerous tissues and adjacent tissues of 50 breast cancer patients.20 normal breast tissue samples were used as control.Results Survivin was expressed in 38 of the 50 breast cancer tissues (76.0%)and in 11 of 50 breast adjacent normal tissues(22.0%)while no expression was found in normal tissues.The positive protein expression of survivin was related to axillary lymph node metastasis of breast cancer while it was not related to expression of PR,ER,Her-2 or ages of patients.Conclusion Survivin may play a role in breast tumorigenesis and is an important predictive parameter in breast cancer patients.
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Objective To investigate the activation of NF-κB and regulation by ubiquitin (Ub)-proteasome pathway in the aorta of rats with chronic renal failure(CRF).Methods The CRF rat model was established by right nephreetomy and left branch renal artery ligation.The CRF rats were were randomly divided into simple CRF group(n=20)and CRF+M used as control group(CON).The NF-κB and the Ub mRNA expression were detected by RT-PCR,and its protein expression was analyzed by immunohistochemistry method.The activity of NF-κBwas mesured by EMSA method.The concentration of IL-1 and TNF-α was detected by ELISA.Results Compared with the CON group,the concentration of serum IL-1 and TNF-α was increased significantly in CRF group [IL-1:(9.02±1.29) vs (2.74±0.96)mg/L,P＜0.01;TNF-α:(50.02±9.52) vs (14.04±1.29)mg/L,P＜0.01]at month 4 after operation.The mRNA expression of NF-κB and Ub in the aorta of CRF group was 1.38 and 1.29 times as that of CON group(P＜0.01).and the protein expression of NF-κB and Ub was 3.75 and 20.5 times as that of CON group(P＜0.01).Compared with the CON group,the activity of NF-κB in the aorta of rats of CRF group was elevated markedly at month 4 after operation(P＜0.01).All the indices were further increased at month 6 after operation.Compared with CRF group,the concentrations of serum IL-1and TNF-α were decreased significantly in CRF+M group[IL-1:(2.94±0.33)mg/L,P＜0.01;TNF-α:(12.80±2.12)mg/L,P＜0.01].The mRNA and protein expression of NF-κB and Ub were also decreased markedly(P＜0.01),and the activity of NF-κB was decreased significantly at month 4 to 6 after operation(P＜0.01).But the amount of ubiquitnative protein was increased significantly in the aorta of CRF+M group as compared to CRF group(P＜0.01). Conclusion The inflammatory signal pathway of ubquitin-proteasome-NF-κB pathway was activated in the aorta of CRF rats,and the proteasome was probablely an important pharmacological intervention target to regulate the activation of NF-κB.
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0 05] Conclusion Ang (1 7) can exert certain protective effect on renal fibrosis in 2K1C hypertensive rats, and can attenuate proteinuria
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Objective To investigate the changes in the expression of inflammatory cytokines in serum and aorta of rats with chronic renal failure.Methods Animal model of chronic renal failure was established by single nephrectomy and partial ligation of the contralateral renal artery.The rats were randomly divided into three groups(10 each):normal control group(Con),chronic renal failure group(CRF),rats with chronic renal failure and treated with proteasome inhibitor MG-132 group(CRF+M).The protein expression of TNF? was detected by immunohistochemistry and image analysis,the levels of TNF? and IL-1? of peripheral blood mononuclear cells(PBMC) in blood plasma and culture supernatant were detected by ELISA.Results The protein expression of TNF? 4 and 6 months after operation significantly increased in aorta of CRF group compared with that in CON group(10.7?1.3 vs 4.3?1.7,12.6?2.2 vs 4.3?1.1,P
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Aim To illustrate the actions and molecular mechanisms of interventions taken to convert the metabolism pathways of cellular apoptosis caused by hypoxia and hypoxia-reperfusion in hypertrophic cardiomyocytes.Methods Angiotensin Ⅱ(0.1 ?mol?L-1)was applied to induce the hypertrophy of mice cardiomyocytes.The cardiomyocytes received the treatment of hypoxia-reperfusion in a tri-gas incubator to simulate the conditions of hypoxia-reperfusion.Before hypoxia/reperfusion,no drug intervention and pre-treatments of DCA(1 mmol?L-1),TMZ(5 ?mol?L-1),LC(50 ?mol?L-1)and AA(10 ?mol?L-1)were given respectively.The glucose and fatty acid oxidative metabolism rates were measured with radioactive counting methods.RT-PCR and Western blot methods were employed respectively to measure the mRNA and protein expression levels of cytochrome C and apoptosis inducers.The spectrophotometry method was used to measure the activity of Caspase-3 and Hoechst 33258 staining to quantify the percentage of cellular apoptosis.Results At post-hypoxia 12 h and post-reperfusion 4 h,the glucose oxidative metabolism rates in hypertrophic cardiomyocytes all decreased while the fatty acid oxidative metabolism rates increased.DCA,TMZ and LC all could inhibit both the reduction of glucose oxidative metabolism after hypoxia-reperfusion and the elevation of fatty acid oxidative metabolism after hypoxia-reperfusion.AA drove the reduction of glucose oxidative metabolism rate even lower and the fatty acid oxidative metabolism rate even higher in hypertrophic cardiomyocytes after ischemia/reperfusion.At the same time,DCA,TMZ and LC could inhibit the expression levels of mitochondrial cytochrome C and AIF mRNA and proteins,the nuclear translocation of cytochrome c and AIF proteins and the activity of caspase-3.And with the opposing actions to AA,DCA,TMZ and LC could inhibit the apoptotic rate of hypertrophic cardiomyocytes after hypoxia-reperfusion.And AA had the opposite effect.Conclusion Intervening in the metabolism pathway of hypertrophic cardiomyocytes was an effective way to prevent and control their programmed death through inhibiting the expression of mitochondrial apoptotic proteins.