1.Effects of mitochondrial fission in proliferation, apoptosis and invasion of thyroid squamous carcinoma cell line SW579
Feifei SHEN ; Rui GUO ; Shupeng ZHAO ; Fengjie QI ; Cuili MENG
Tianjin Medical Journal 2015;(2):129-132,133
Objective To detect the expression of mitochondrial dynamics proteins (Mfn2 and Drp1) in thyroid squa?mous carcinoma cell line SW579 and the effects of Mitochondrial division inhibitor, Mdivi-1, on proliferation, apoptosis and invasion of SW579. Methods In SW579 and Nthy-ori 3-1 cell lines, the expression levels of Mfn2 and Drp1 were deter?mined by western blot while the transcription level of Mfn2 and Drp1 mRNA were measured by RT-PCR. Then, SW579 cells were divided into control group (DMSO, 0.1%) and Mdivi-1 low, medium and high dose groups (Mdivi-1 of 15,30 and 45μmol/L were incubated with cells for 16 hours respectively). Then the ability of cell proliferation was detected using MTT assay, the mitochondrial membrane potential was determined by fluorescence spectrophotometer, the expression levels of cy?tochrome C and Caspase-3 were quantified by Western blot and the transcription level of the Cyt C and Caspase-3 mRNA were determined by RT-PCR. The ability of invasion in each group was measured with Transwell assays. Results Com?pared with Nthy-ori 3-1, the mRNA transcription and protein expression levels of the Mfn2 was remarkably decreased, while the mRNA transcription and protein expression of the Drp1 was significantly increased in SW579 cells (P<0.01). Compared with control group, the cell survival rates and mitochondrial membrane potential of SW579 were decreased dramat?ically (P<0.01). The mRNA transcription and protein expression of the cytochrome C and Caspase-3 were increased dra?matically (P<0.01) and the capability of invasion was markedly decreased in all the Mdivi-1 groups in a dosage dependent manner compared with those in control groups (P<0.01). Conclusion Abnormal mitochondrial dynamics may be involved in thyroid squamous cell carcinoma SW579 cells;Mdivi-1 can inhibit the cell proliferation and invasion as well as induce apoptosis.
2.Effects of curcumin on TGF-β1 and smad7 expression in rat renal tubular epithelial cells stimulated with AngⅡ
Yuxia WANG ; Fang WANG ; Feifei SHEN ; Huasheng CUI ; Huimin WANG
International Journal of Traditional Chinese Medicine 2017;39(1):39-42
Objective To observe the effect of curcumin on the expression of smad7 and TGF-β1 in rats renal tubular cells of with Ang II, and discuss the mechanism of curcumin to improve renal interstitial fibrosis.Methods Cultured rat renal tubular epithelial cells were divided into the blank group, the Ang II control group and the low, medium and high dose curcumin group. The rest of the groups were intervened by 10-8 mol/L Ang II except the blank group; the low, medium and high dose groups of curcumin were intervened by 2.5, 5.0, 10.0μmol/L curcumin. Then Western blot was used to detect the expression of TGF-β1 and smad7 protein, RT-PCR was used to detect the TGF-β1 and smad7 mRNA expression.Results Compared with the blank group, the expression of TGF-β1 protein (0.23 ± 0.03vs. 0.16 ± 0.01), and TGF-β1 mRNA (1.89 ± 0.20vs. 1.00 ± 0.00) significantly increasedin AngⅡ control group (P<0.05), and the expression of smad7 protein (0.19 ± 0.03vs. 0.24 ± 0.02), and smad7 mRNA (0.48 ± 0.05vs. 1.00 ± 0.00) significantly reduced in AngⅡcontrol group (P<0.05). Compared with the AngⅡ control group, the expression of TGF-β1 protein in low, medium and high dose curcumingroup (0.18 ± 0.02, 0.17 ± 0.02, 0.16 ± 0.03vs. 0.23 ± 0.03) and TGF-β1 mRNA (1.58 ± 0.11, 1.34 ± 0.16, 0.97 ± 0.19vs. 1.89 ± 0.20) significantly decreased (P<0.05), and the expression of smad7 protein (0.28 ± 0.04, 0.31 ± 0.03, 0.34 ± 0.04vs. 0.19 ± 0.03) and smad7 mRNA (0.68 ± 0.07, 0.80 ± 0.06, 0.98 ± 0.09vs.0.48 ± 0.05) increased significantly (P<0.05).Conclusions Curcumin can thus play its role in renal protection by counteract the AngⅡ mediated renal interstitial fibrosis. Its mechanism may be related to the reduction of TGF-β1 protein and its mRNA expression, up regulation of smad7 protein and its mRNA expression.
3.Expression of p53 gene in human embryonic lung fibroblasts induced by sodium arsenite
Yan ZOU ; Xubo SHEN ; Hui JIANG ; Feifei JIA ; Yungang XIONG
Medical Journal of Chinese People's Liberation Army 1982;0(01):-
Objective To study the dose-dependent effect of sodium arsenite on the expression of p53 gene in human embryonic lung fibroblasts(HELF)in vitro.Methods The human embryonic lung fibroblasts were divided into four groups in vitro based on completely randomized design.The expression of p53 mRNA was detected by real-time PCR and the expression of p53 protein was detected by immunohistochemical SAB,respectively,in human embryonic lung fibroblasts which were exposed to different doses(0,3,9 and 15?mol/L)of sodium arsenite for 24 hours.The one-way analysis of variance and post hoc comparisons were performed for testing the differences among groups and the linear correlation was for testing correlation between doses and the expression of p53 gene.Results Compared with that of exposure to 0?mol/L sodium arsenite,the expression of p53 mRNA in HELF was increased significantly(P0.05).The p53 mRNA expression was increased in dose-dependent manner with the increased concentration of sodium arsenite(r=0.947,P
4.Expression and distribution of N-myc downstream regulated gene 2 in pancreatic diseases
Feifei HE ; Kai JIAO ; Xinping LIU ; Lan SHEN
Chinese Journal of Endocrinology and Metabolism 2013;(2):162-164
The pancreatic tissues from patients with islet cell hyperplasia,insulinoma,and pancreatic adenocarcinoma,as well as normal pancreatic tissues were embedded with paraffin,serial sections were cut and mounted on glass slides.Immunohistochemical staining was carried out with N-myc down-stream regulated gene 2 (Ndrg2) monoclonal antibody by means of ABC method,and Western blotting was carried out to detect the expression and distribution of Ndrg2.The results showed that Ndrg2 positive immunoreactivity was mainly localized in the cytoplasm of islet cell,being similar to the localization of insulin positive immunoreactivity.The number and volume of pancreatic islets were increased in the patients with islet cell hyperplasia,and Ndrg2 expression was also increased.Western blotting results showed that the expression of Ndrg2 in the pancreas of patients with islet cell hyperplasia was increased compared with normal group.The above results suggest that Ndrg2 may play an important role in performing physiological function of islet cells.
5.Changes in serum TGF ?_1 in type 2 diabetic patients
Haibing JU ; Peizhen GAN ; Jing LI ; Feifei SHEN ; Zizhen SHU
Chinese Journal of Pathophysiology 2000;0(11):-
AIM: To explore the changes in serum TGF ? 1 in type 2 diabetes mellitus. METHODS: Forty-five cases type 2 diabetes mellitus patients were divided into three groups according to urine albumim excretion rate(UAER): normoalbuminuria(NA)group and microalbuminuria(MA) group and macroalbuminuria group (Overt DN). Serum TGF ? 1, fasting blood glucose(FBG), HbA 1c ,BUN,Cr,Ccr,lipidemia were detected in all cases. RESULTS: Serum TGF ? 1 in NA, MA and ODN groups [(35.02?6.70) ?g/L, (39.31?5.35) ?g/L, (58.58?9.56) ?g/L, respectively] was higher than that in control [(23.95?8.01) ?g/L, P
6.Effects of exenatide on the cell proliferation, invasion and apoptosis of human tongue squamous cell carcinoma SCC-25
Chao HUANG ; Feifei SHEN ; Gang LI ; Yue ZHAO
Tianjin Medical Journal 2015;43(5):461-464
Objective To detect the effects of exenatide on the related indicators of proliferation, invasion and apoptosis of cell line SCC-25. Methods SCC-25 cells were cultured in vitro. The expression level of glucagon like peptide 1 receptor (GLP-1R) was determined by Western blot assay in SCC-25 cells. SCC-25 cells were divided into four groups:control group and exenatide group (1,10 and 100 nmol/L). The ability of cell proliferation was detected using MTT assay after 24 h, 48 h and 72 h of culture. The ability of invasion was measured with Transwell assays. The expression levels of MMP-2, Caspase-3 and Phospho-p38 MAPK were measured by Western blot assay. Results GLP-1 receptor expression was found in SCC-25 cells. Compared with control group, the cell survival rate, invasion rate and the expression of MMP-2 were significantly decreased in SCC-25 group (P<0.05). The expression of Caspase-3 were significantly increased (P<0.05).Changes were in a concentration-dependent and time-dependent manner (P<0.05). The expression of Phospho-p38 MAPK was significantly increased at 24 h in 10 nmol/L exenatide group (P<0.05). Conclusion Exenatide can inhibit the cell proliferation and invasion, which may contribute the apoptosis by promoting expressions of Phospho-p38 MAPK and Caspase-3 of SCC-25 cells.
7.Effects of different doses of topiramate on expression of NCAM and GAP- 43 mRNA in hippocampus of rats with epilepsy
Danhong WU ; Ping ZHONG ; Guanshui BAO ; Hong ZHOU ; Jun SHEN ; Feifei HUANG
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(12):1459-1462
Objective To explore the effects of different doses of topiramate (TPM) on the expression of nerve cell adhesion molecule (NCAM) and growth-associated protein 43 (GAP- 43) mRNA in hippocampus of rats with epilepsy. MethodsForty-eight rats were randomly divided into normal control group, kainic acid (KA) group, 10 mg/kg TPM group, 40 mg/kg TPM group, 100 mg/kg TPM group and 400 mg/kg TPM group (n=8). The models of rats with epilepsy treated by different doses of TPM were established. The behavior of rats was observed, and the expression of NCAM and GAP- 43 mRNA in hippocampus of rats was determined by Real-time PCR. Results The expression of NCAM and GAP- 43 mRNA in KA group was significantly higher than that in normal control group (P<0.01), while there was no significant difference between 10 and 40 mg/kg TPM groups and KA group, that in 100 and 400 mg/kg TPM groups was significantly lower than that in KA group (P<0.01), and that in 400 mg/kg TPM group was significantly lower than that in 100 mg/kg TPM group (P<0.01). Conclusion KA can up-regulate the expression of NCAM and GAP- 43 mRNA in hippocampus of rats with epilepsy. Higher dose of TPM can inhibit the expression of NCAM and GAP- 43 mRNA, and the inhibitory effect is related with the dose of TPM.
8.Study on protection of Bcl-2 on islet cells cultured in immunosuppressive drugs
Haibing JU ; Zizheng SHU ; Dengming HU ; Huiying YING ; Feifei SHEN ; Ziying ZHU
Journal of Chinese Physician 2001;0(01):-
Objective To investigate the toxic effect of immunosuppressive drugs on apoptosis of rat pancreatic islet cells in vitro and the protective action of Bcl-2.Methods Islet cells expressing Bcl-2 and the control islet cells were cultured at different concentrations of tacrolimus and the apoptosis rate of islet cells and insulin accumulation in the culture medium were detected after 48h.Results Low and high concentrations of tacrolimus induced the apoptosis of islet cells and decreased insulin secretion.The Bcl-2 inhibited the apoptosis of islet cells induced by tacrolimus and improved the insulin secretion.Conclusion Tacrolimus may directly damage to isolated rat islet cells and the expression of Bcl-2 can protect the cells from the damage of immunosuppressive drugs.
9.Effects of antithyroid drug on oxidative stress in Graves ophthalmopathy patients
Haibing JU ; Guanglin WANG ; Dengming HU ; Zizheng SHU ; Ziying ZHU ; Feifei SHEN
Journal of Chinese Physician 2001;0(08):-
Objective To investigate the effects of antithyroid drug(ATD) on oxidative stress in Graves disease patients with and without infiltrative ophthalmopathy.Method The levels of superoxide dismutase(SOD),glutathione peroxidase(GPx) and catalase(CAT) in erythrocytes of Graves disease patients(18 with and 20 without infiltrative ophthalmopathy) were measured before and after treatment with ATD.Results Compared with the normal control group,the levels of SOD,GPx and CAT were significantly higher in Graves disease patients with and without infiltrative ophthalmopathy.After euthyroidism was achieved with the treatment of ATD,the levels of SOD,GPx and CAT were normalized in the patients without infiltrative ophthalmopathy,but oxidative stress was still present in the patients with infiltrative ophthalmopathy.Conclusion The oxidative stress is obvious in patients with Graves ophthalmopathy,which suggests that oxidative stress is involved in orbital inflammation.
10.Neuroimaging study of CADASIL pedigree with performance of familial migraine
Hong CHENG ; Qingwen JIN ; Qi NIU ; Feifei SHEN ; Juan YAO ; Xinsheng DING
Chinese Journal of Nervous and Mental Diseases 2015;(9):542-546
Objective To analysis the MRI features of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), to improve the understanding of MRI manifestations of this disease. Meth?ods The clinical manifestations, neuroimaging analysis and genetic analysis were performed in the CADASIL pedigree proband and his families. Results Five of six cases were confirmed with C2182T mutation on exon 14 of the NOTCH3, of which three cases were diagnosed by MRI. Brain MRI findings included bilateral symmetric distributed confluent lesions in the subcortical and periventricular white matter in the frontal lobe, hypointensity on T1WI and hyperintensity on both T2WI and T2 FLAIR imaging in four cases. The external capsule was involved in three cases, with hyperintensity on T2WI. Subcortical lacunar lesions (SLLs) were shown in three cases. Lacunar infarction in the basal ganglia and thalamus were presented in four cases. T2WI hyperintensity at the brain stem was found in two cases. Cerebral microbleeds were re?vealed in three cases. There was no O’Sullivan sign in all the six cases. Conclusions There is characteristic change of MRI in CADASIL patients, which may play a very important role in screening these cases.