1.WNK4 kinase-mediated inhibitory effect on expression of BK channel via lysosomal pathway
Jieqiu ZHUANG ; Dexuan WANG ; Yiqian ZHANG ; Weihui NIU ; Fangxuan CHEN ; Zhen SHI ; Shufang PAN ; Dingying GU
Chinese Journal of Nephrology 2012;28(4):291-295
Objective To investigate the mechanism underlying the WNK4 kinasemediated inhibitory effect on BK channel. Methods Cos-7 cells were cotransfected with BK in combination with either CD4 (control group) or wild type WNK4 (WNK4-WT).Immunostaining and confocal microscopy,chemiluminescence,Western blotting analysis were then employed to determine the BK localization in cells,BK surface expression and total protein level,respectively.To further investigate whether the reduction of BK protein expression is due to an increase in degradation through a lysosomal pathway,BK protein level was determined after treated with bafilomycin A1(Baf A1),a proton pump inhibitor affecting lysosomal degradation. Results Immunostaining and confocal microscopic study showed that BK was localized both in plasma membrane and cytosol in the control group.After cells transfected with WNK4-WT,BK expression was markedly reduced.Chemiluminescent assay found that BK surface expression level was 299.9±18.6 in the control group,whereas it was significantly reduced (148.4±13.7,P<0.01) in the WNK4-WT group.Western blotting analysis showed that total BK protein level was markedly reduced in the presence of WNK4-WT compared to the control group.WNK4-WT was shown to significantly reduce the BK total protein level (42.3%±15.2%) compared to the control group (100%) (P<0.01).When the cells was treated with Bafilomycin A1 (Baf A1,0.5 μmol/L),WNK4-mediated reduction in BK protein was reversed (82.2%±12.1%,P<0.05). Conclusions WNK4 inhibits total and surface protein expression of BK in Cos-7 cells whick is likely due to an increase in BK degradation through a lysosomal pathway.
2.Effects of NS398, an inhibitor of cyclooxygenase 2, on the growth and apoptosis of human squamous cell carcinoma cell line Tca8113
Weihui ZENG ; Yan ZHENG ; Songmei GENG ; Jingyi YUAN ; Xinwu NIU ; Shengshun TAN
Chinese Journal of Dermatology 2009;42(1):42-44
Objective To study the effect of NS398, an inhibitor of cyclooxygenase 2 (COX2), on the growth and apoptosis of human squamous cell carcinoma cell line Tca8113. Methods Cultured Tca8113 cells were incubated with NS398 (0, 6.25, 12.5, 25, 50, 100 μmol/L) for 24, 48 and 72 hours, respectively. Thereafter, MTT method, flow cytometry and transmission electron microscopy were applied to detect the proliferation, cell cycle and apoptosis of Tca8113 cells, respectively. Results The proliferation of Tca8113 cells was inhibited by NS398 in a dose- and time-dependent manner (both P<0.05). FCM analysis showed the appearance of a typical hypodiploid apoptotic (Sub-G1) peak, an increase in the percentage of cells at G0/G1 phase and a decrease in that at S and G2/M phases in NS398 ( 100 μmol/L) -treated Tca8113 cells. Moreover, the cell proliferation index was significantly downregulated by NS398 of 100 μmol/L from 41.03 to 24.33 (P<0.05). Under an electron microscope, morphological changes characteristic of apoptosis were observed in NS398-treated Tca8113 cells. Conclusion NS398, an inhibitor of COX2, could effectively inhibit the growth of Tca8113 cells in vitro by induction of apoptosis.
3.Effects of Notch signal on the expressions of HIF-α and autophagy- related genes Beclin1, LC3I, LC3II in oxygen-glucose deprivation induced myocardial cell injury.
Ling Yu KONG ; Zan XI ; Wen Ting MA ; Fei Yun YANG ; Li Dan NIU ; Jin He SHI
Chinese Journal of Applied Physiology 2019;35(2):165-168
OBJECTIVE:
To investigate the effects of Notch signal on hypoxic induction factor (HIF-1α) and autophagy-associated genes Beclin1, LC3I, LC3II in oxygen-glucose deprivation (OGD) induced myocardial cell injury.
METHODS:
The OGD model was established using hypoxic culture box and hypoglycemic DMEM medium. The cells were divided into normal control group, OGD group, OGD + NC siRNA group, OGD + Notch1 siRNA group and OGD + HIF-1α siRNA group. Western blot was used to detect the interference effects of HIF-1α siRNA and Notch1 siRNA. The effects of Notch1 siRNA and HIF-1α siRNA on the activity of myocardial cells in OGD model were detected by the CCK-8 assay. The effects of Notch1 siRNA and HIF-1α siRNA on autophage-associated genes Beclin1, LC3I and LC3II expression were detected by Western blot.
RESULTS:
The results of Western blot showed that HIF-1α siRNA could effectively knock down the expression of HIF-1α in myocardial cells in OGD model, and Notch1 siRNA could effectively knock down the expression of Notch1 and HIF-1α in myocardial cells in OGD model. The result of CCK-8 assay showed that Notch1 siRNA and HIF-1α siRNA reduced the activity of myocardial cells in OGD model, and there was no statistical difference between the two groups. Western blot results showed that Notch1 siRNA and HIF-1α siRNA could reduce the expressions of the autophagy-associated genes Beclin1, LC3I and LC3II, and reduce the ratio of LC3II to LC3I at mRNA level.
CONCLUSION
Notch1 plays a role in myocardial protection by regulating the expression of HIF-1α to regulate the autophagy in OGD model cells.
Autophagy
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Beclin-1
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metabolism
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Cell Hypoxia
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Cells, Cultured
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Glucose
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Humans
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Hypoxia-Inducible Factor 1, alpha Subunit
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metabolism
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Microtubule-Associated Proteins
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metabolism
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Myocytes, Cardiac
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cytology
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pathology
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Oxygen
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Receptors, Notch
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metabolism
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Signal Transduction
4.Changes in corneal epithelial thickness and optical density and their correlation after smart pulse technology-assisted transepithelial photorefractive keratectomy
Shiyang NIU ; Hua YANG ; Yan LI ; Zhiqiang DAI ; Xinmin LI ; Yulan ZHOU ; Ouyang ZHANG ; Baojun WANG
International Eye Science 2024;24(8):1308-1313
AIM: To evaluate the changes in corneal epithelial thickness(CET)and corneal optical density(CD)after smart pulse technology(SPT)-assisted transepithelial photorefractive keratectomy(TPRK)and analyze their correlation.METHODS: The prospective study included 60 patients(120 eyes)with myopia and myopic astigmatism who underwent SPT-TPRK in the ophthalmology department at the First Affiliated Hospital of Xinxiang Medical University between February and August 2023. Changes in CET and CD were evaluated preoperatively and at 1 wk, 1 and 3 mo postoperatively.RESULTS: A total of 14 cases(28 eyes)were lost to follow-up, and 3 patients(6 eyes)with postoperative haze were excluded from this study, resulting in a final inclusion of 43 patients(86 eyes). At 1 wk after SPT-TPRK, CET had statistically significantly thickened compared to preoperative levels(P<0.05), particularly in the CET at 0-2 mm central corneal area(P<0.05). At 1 mo after SPT-TPRK, the CET at 0-2 mm area had statistically significantly decreased(P<0.05). At 3 mo after SPT-TPRK, the CET at 0-2 mm had essentially reached preoperative levels. Postoperative CD values increased, with a positive correlation between CET in the 0-2 mm area and CD in the whole 0-2 mm area(r=0.256, P<0.05), and a positive correlation between CET in the 2-5 mm area and CD in the anterior 2-6 mm area(r=0.319, P<0.05).CONCLUSION: Corneal epithelial remodeling takes 3 mo in areas within 2 mm of the central cornea; areas with thinner CET have faster postoperative corneal epithelial remodeling and greater thickening in the early postoperative period; CD increases in the early postoperative period compared to the preoperative value, and in some areas, there is a positive correlation between CET and CD value.
5.Effect of traditional chinese medicine on survival and quality of life in patients with esophageal carcinoma after esophagectomy.
Ping LU ; Qiu-dong LIANG ; Rong LI ; Hong-rui NIU ; Xiao-ge KOU ; Hong-jun XI
Chinese journal of integrative medicine 2006;12(3):175-179
OBJECTIVETo explore the effect and possible mechanism of traditional Chinese medicine (TCM) on survival and quality of life (QOL) in patients with esophageal carcinoma after esophagectomy.
METHODSAdopting prospective controlled method of study, the authors had 128 post-esophagectomy patients, hospitalized from February 2001 to February 2002, randomly divided into 3 groups: the TCM group, treated with TCM drugs alone; the chemotherapy group, with chemotherapy alone applied; and the synthetic group, treated with chemotherapy combined with Chinese medicine. Their survival rate and QOL were compared.
RESULTSIn the TCM group, the chemotherapy group and the synthetic group, the respective 3-year relapse and remote metastasis rate were 71.4%, 76.7%, 53.4%, respectively (chi(2) = 6.53, P < 0.05); the 1-year survival rate 42.9%, 46.5%, 72.1%; 2-year survival rate 28.6%, 27.9%, 55.8%, and 3-year survival rate 26.2%, 23.1%, 37.2%, respectively. And the QOL improving rate was 69.0%, 37.2%, 58.1%, respectively, all showing significant difference among them (chi(2) = 6.10, all P < 0.05). Moreover, immune function was increased in the TCM and the synthetic groups.
CONCLUSIONIntegrative Chinese and Western medicinal treatment was the beneficial choice for post-operational patients with esophageal carcinoma. However, long time use of simple Chinese medicine was also advisable, especially for those in poverty.
Adult ; Aged ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Esophageal Neoplasms ; drug therapy ; mortality ; surgery ; Esophagectomy ; Female ; Follow-Up Studies ; Humans ; Immune System ; drug effects ; Immunoglobulins ; blood ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; mortality ; Prospective Studies ; Quality of Life ; Survival Rate ; T-Lymphocyte Subsets