1.Advanced oxidation protein products modulate autophagy of renal tubule epithelial cells
Jun ZHANG ; Xiaohong XIANG ; Xiujie LIANG ; Shuangshuang SHU ; Tingting JIANG ; Tingting GUO ; Xun TANG
Chongqing Medicine 2017;46(25):3480-3483
Objective To investigate the effect of advanced oxidation protein products (AOPPs) on human renal tubular epithelial cells(HK-2) autophagy.Methods HK-2 cells were stimulated with AOPPs.RT-qPCR and Western blot were used to determine the expression of autophagy related protein LC3-Ⅱ/LC3-Ⅰ,Beclin1 and p62;Western blot was utilized to examine the activation of p38 MAPK pathway.Then p38 MAPK inhibitor (SB203580) was added and co-processed with AOPPs.The change of autophagy was observed Also,autophagy inducer rapamycin was added and co-processed with AOPPs.RT-qPCR and Western blot were used to detect the expression of cell cycle inhibitory protein p27.The cell total protein level was detected by the bicinchoninic acid (BCA) method.The hypertrophy change was observed.Results AOPPs down-regulated the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin1,up-regulated expression of p62 and activated p38 MAPK pathway;in comparison with the AOPPs alone treatment group,the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin in the SB203580 co-processing group was increased,while p62 was decreased;the p27 expression and cells total protein in the sirolimus co-processing group were down-regulated.Conclusion AOPPs inhibits the autophagy of HK-2 cells by activating p38 MAPK pathway and the autophagy inhibition participates in HK-2 cell hypertrophy.
2.Wnt3a is Important in The Differentiation From Neural Stem Cell Into Dopaminergic Neuron In vitro
Shu HAN ; Wei SHI ; Yanhua LI ; Hailei YAO ; Xiaoyan XIE ; Lin CHEN ; Shuangshuang SHI ; Cixian BAI ; Xue NAN ; Fang YAN ; Yunfang WANG ; Xuetao PEI
Progress in Biochemistry and Biophysics 2006;0(10):-
Wnt signaling is implicated in the control of cell growth and differentiation during neural stem cell(CNS) development.Wnt3a, one of wnt gene family members, has effect on regeneration neurospheres and differentiation into neurons.Wnt3a inhibits regeneration of neurospheres, and promotes its differentiation. In vitro neurosphere was cultured in a serum-free defined medium DMEM/F12 supplemented with bFGF and EGF. Dissociated cells were plated onto poly-d-lysine-coated coverslips and propagated in medium containing recombined Wnt3a-adenovirus. Plenty of Nurr1 were detected by RT-PCR after 3 days. Wnt3a combined AA would improve NSC differentiation into dopaminergic (DA) neuron. The quantity of DA neuron is obviously more than the AA alone group's. Moreover, the expression of TH mRNA is 1.86 fold in Wnt3a combined AA group. Induced cells were immunostained for TH and DAT. The proportion of TH-positive was (37.42 ? 2.54) % (P
3.Effect of preoperative pulmonary hypertension on right ventricular function of cardiac allograft assessed by two-dimensional speckle tracking echocardiography
Xin ZHANG ; Shuangshuang ZHU ; Wenqian WU ; He LI ; Lingyun FANG ; Manwei LIU ; Yi ZHOU ; Yang ZHAO ; Li ZHANG ; Yuman LI ; Guohua WANG ; Shu CHEN ; Nianguo DONG ; Mingxing XIE
Chinese Journal of Ultrasonography 2022;31(12):1013-1020
Objective:To investigate the effect of preoperative pulmonary hypertension (PH) on right ventricular function in patients with heart transplantation(HTx) one year after surgery.Methods:A total of 120 patients who underwent HTx in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2017 to January 2020 were retrospectively recruited.According to the mean pulmonary arterial pressure (mPAP) obtained by preoperative right heart catheterization, the research subjects were divided into the pulmonary hypertension group (PH group, n=81) and without pulmonary hypertension group (NPH group, n=39). Conventional echocardiographic indices of right ventricular function such as right ventricular area change (RV-FAC), tricuspid annular plane systolic excursion (TAPSE), tricuspid lateral annular systolic velocity (S′), and two-dimensional speckle tracking imaging (2D-STI) strain parameters including right ventricular global longitudinal strain (RV-GLS), right ventricular free wall longitudinal strain (RV-FWLS) were obtained to assess the right ventricular function of grafted hearts. The echocardiographic parameters one year after the operation of the two groups were analyzed to compare the differences in right ventricular function and their correlation with preoperative mPAP. Results:The grafted heart RV-GLS and RV-FWLS were significantly decreased in the PH group (all P<0.01), while RV-FAC, TAPSE, and S′ were similar between the two groups (all P>0.05). RV-FWLS and RV-GLS correlated with preoperative hemodynamic parameter mPAP( rs=-0.46, -0.54; all P<0.05)while RV-FAC, TAPSE, and S′ were not significantly correlated with mPAP (all P>0.05). Conclusions:Preoperative PH correlates with right ventricular function in HTx patients 1 year after the operation. The absolute values of RV-FWLS and RV-GLS in HTx patients with preoperative PH decrease 1 year after the operation. 2D-STI is more sensitive than conventional echocardiography to monitor the changes in right ventricular function in HTx patients after the operation.
4.Clinical Efficacy of Qihuang Jianpi Zishen Granules in Treating Cardiac Involvement in Systemic Lupus Erythematosus
Yunfei LI ; Lijun PANG ; Longwu SHU ; Wanlan FU ; Shuangshuang SHANG ; Ming LI ; Chuanbing HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(22):104-111
ObjectiveTo investigate the clinical efficacy and safety of Qihuang Jianpi Zishen granules in the treatment of cardiac involvement in systemic lupus erythematosus (SLE). MethodA total of 62 SLE patients with cardiac involvement treated in the Department of Rheumatology and Immunology, the First Affiliated Hospital of Anhui University of Chinese Medicine from June 2021 to December 2022 were randomized into control and observation groups (n=31). The control group was treated with methylprednisolone tablets and hydroxychloroquine sulfate tablets, and the observation group with Qihuang Jianpi Zishen granules on the basis of the therapy in the control group. After 12 weeks of treatment, the two groups were compared in terms of the therapeutic effect, cardiac function indicators [left atrial end-diastolic diameter (LADd), left ventricular end-diastolic diameter (LVDd), left ventricular posterior wall thickness (LVPWTd), peak blood flow velocity in early diastolic period (peak E), peak blood flow velocity in late diastolic period (peak A), E/A ratio, stroke volume (SV), left ventricular ejection fraction (LVEF), left ventricular short axis shortening rate (LVFS), and B-type natriuretic peptide (BNP)], vascular damage indicators [nitric oxide (NO), endothelin-1 (ET-1), vascular endothelial growth factor (VEGF), and homocysteine (Hcy)], inflammation indicators [erythrocyte sedimentation rate (ESR) and hypersensitive C-reactive protein (Hs-CRP)], anti-double-stranded DNA (dsDNA) antibodies, disease activity index (SLEDAI) score, mitigation of symptoms and signs, and occurrence of adverse reactions. ResultThe total response rate in the observation group was 87.09%, which was higher than that (67.74%) in the control group (P<0.01), and the incidence of adverse reactions had no significant difference between the two groups. After treatment, the control group showed lowered LVDd, LVPWTd, BNP, ET-1, VEGF, and Hcy (P<0.05) and increased E peak, E/A ratio, SV, LVEF, and LVFS (P<0.05). In the observation group, LADd, LVDd, LVPWTd, peak A, BNP, NO, ET-1, VEGF, and Hcy decreased (P<0.05), while peak E, E/A ratio, SV, LVEF and LVFS increased (P<0.05) after treatment. The treatment in both groups decreased the scores of palpitation, chest tightness, dyspnea, and edema (P<0.05), reduced ESR, Hs-CRP, ds-DNA, and SLEDAI (P<0.05). After treatment, the observation group had lower LADd, LVDd, LVPWTd, peak A, BNP, and scores of palpation, chest tightness, dyspnea, and edema (P<0.05) and higher peak E, E/A ratio, SV, LVEF, and LVFS (P<0.05) than the control group. In addition, the observation group had lower NO, ET-1, VEGF, Hcy, ESR, Hs-CRP, ds-DNA, and SLEDAI than the control group (P<0.05). ConclusionQihuang Jianpi Zishen granules combined with hydroxychloroquine sulfate and methylprednisolone can improve multiple indicators and mitigate the symptoms and signs of SLE patients with cardiac involvement, demonstrating a clinical application value.
5.Mechanism of cinobufagin regulating PI3K/AKT signaling pathway to reverse cisplatin resistance in ovarian cancer A2780/DDP cells
Meiling Shu ; Yue Wu ; Yingquan Ye ; Shuangshuang Zhang ; Mei Zhang
Acta Universitatis Medicinalis Anhui 2024;59(4):671-677,741
Objective :
To investigate the reversal effect and mechanism of cinobufagin (CBG) on cisplatin resist- ance in human ovarian cancer cells .
Methods :
A2780 cell line and its cisplatin-resistant cell line A2780/DDP are common ovarian cancer cells in clinic , so these two cell lines were selected as the research objects . The cell viabil- ity was detected by cell Counting Kit-8 (CCK-8) assay , and the cell proliferation ability was detected by plate clo- ning and 5-ethynyl-2 ′-deoxyuridine (EdU) assay. Hoechst staining was used to observe cell apoptosis . Cell scratch test and Transwell test were used to evaluate cell migration and invasion ability. Western blot and quantitative reverse transcription PCR (RT-qPCR) were used to detect the protein and mRNA expressions of phosphatidylinosi- tol 3-kinase/protein kinase ( PI3K/AKT) signaling pathway and epithelial-mesenchymal transition ( EMT) .
Results:
Compared with A2780 cells , the drug resistance indexes of A2780/DDP cells were 5 . 636 , 5 . 864 , 5 . 695 , respectively. After treatment of A2780/DDP cells with CBG (2 , 4 , 6 mg/ml) , the reversal resistance indexes were 1 . 617 , 2. 570 , 3 . 461 , respectively. CBG treatment significantly increased the level of apoptosis and inhibi- ted the proliferation , migration and invasion of the cells in a concentration-dependent manner (P < 0. 05) . Western blot results showed that compared with A2780 cells , the relative ratio of P-PI3K/PI3K and P-AKT/AKT protein levels , as well as the protein expression of N-cadherin , Vimentin , and Snail were higher in the control group (A2780/DDP) cells , while the protein expression of E-cadherin was lower ( t P-PI3K/PI3K = 8 . 115 , t P-AKT/AKT = 17. 62 , t N-cadherin = 6. 126 , t Vimentin = 4. 001 , t Snail = 17. 333 , t E-cadherin = 4. 620 , P < 0. 01) ; As the dose of CBG increased , the protein expression levels of P-PI3K , P-AKT , N-cadherin , Vimentin , and Snail in drug-resistant cells de- creased , while the protein expression level of E-cadherin increased ( FP-PI3K = 268. 5 , FP-AKT = 190. 5 , FN-cadherin = 24. 02 , F Vimentin = 57 . 65 , FSnail = 87 . 24 , FE-cadherin = 135 . 8 , P < 0. 05) . qRT-PCR results showed that with the in- crease of CBG concentration , the mRNA expression levels of PI3K , AKT , N-cadherin , Vimentin and Snail de- creased , while the mRNA expression level of E-cadherin gradually increased ( FPI3K = 101 . 1 , FAKT = 558. 3 , FN-cadherin = 86. 97 , F Vimentin = 105 . 9 , FSnail = 85 . 71 , FE-cadherin = 80. 96 , P < 0. 01) .
Conclusion
CBG can reverse cisplatin resistance of ovarian cancer A2780/DDP cell line , and its mechanism may be related to the regulation of PI3K/AKT signaling pathway and inhibition of EMT by CBG.