1.Effect of LncRNA RMRP on ferroptosis induced by oxygen glucose deprivation/reperfusion in mouse HL-1 cardiomyocytes by regulating miR-766-5p
Lei He ; Xinglan Sun ; Yingxing Wu ; Yuan Xu ; Xiang Peng ; Chenkai Hu
Acta Universitatis Medicinalis Anhui 2025;60(12):2207-2214
Objective:
To investigate the effect and mechanism of long non-coding RNA RMRP(LncRNA RMRP) on oxygen-glucose deprivation/reperfusion(OGD/R)-induced ferroptosis in mouse HL-1 cardiomyocytes by regulating miR-766-5p.
Methods:
HL-1 cells were cultured in vitro, and OGD/R models were established. The expression levels of LncRNA RMRP in HL-1 cells at various reperfusion time points were subsequently quantified using qRT-PCR. The LncRNA RMRP small RNA interference fragment(si-RMRP) and its corresponding negative control(si-NC), as well as the miR-766-5p inhibitor and its respective negative control(inhibitor-NC), were transfected into HL-1 cells. Subsequently, the cells were subjected to OGD/R treatment. CCK-8 assay was employed to evaluate cell viability. Assay kits were employed to measure the levels of lactate dehydrogenase(LDH) in the cell supernatant, as well as the intracellular levels of malondialdehyde(MDA), superoxide dismutase(SOD), glutathione(GSH), and ferrous ion(Fe2+). qRT-PCR analysis was conducted to assess the expression levels of LncRNA RMRP and miR-766-5p. Western blot analysis was conducted to assess the expression levels of proteins associated with ferroptosis including GPX4, SLC7A11, and FTH1. Dual-luciferase reporter assays were performed to investigate the sponge adsorption relationship between LncRNA RMRP and miR-766-5p.
Results:
As reperfusion time extended, the expression level of LncRNA RMRP in cells progressively increased(P2+ levels within the cells, and decreased the activities of SOD and GSH in cells(P2+ levels within the cells, and promoting SOD and GSH activities in cells(PP < 0. 01) . Furthermore , silencing LncRNA RMRP upregulated the protein expression levels of GPX4 , SLC7A11 , and FTH1(P < 0. 01) . The dual-luciferase reporter assay confirmed that LncRNA RMRP could regulate the expression of miR-766- 5p through a sponge adsorption mechanism. Partial inhibition of miR-766-5p inhibitor expression could mitigate the improvement effect caused by LncRNA RMRP silencing on OGD/R-induced ferroptosis in HL-1 cells.
Conclusion
Silencing LncRNA RMRP inhibits OGD/R-induced ferroptosis in HL-1 cells , potentially through the sponge-mediated regulation of miR-766-5p expression.
2.Effect of chidamide combined with matrine on proliferation and apoptosis of cutaneous T-cell lymphoma cell lines HH and Hut78 and possible apoptotic mechanisms
Xinglan HE ; Yimeng WANG ; Guanyu WANG ; Chunlei ZHANG
Chinese Journal of Dermatology 2020;53(2):121-127
Objective To evaluate the effect of chidamide combined with matrine on proliferation and apoptosis of cutaneous T-cell lymphoma (CTCL) cell lines HH and Hut78,and to explore their apoptotic mechanisms.Methods Both HH and Hut78 cells were treated with 0.4 μmol/L chidamide and 0.6 g/L matrine alone or in combination for 24,48 and 72 hours,with those treated with dimethyl sulfoxide (DMSO) serving as control groups.MTS assay was performed to deteet cellular proliferation rates of HH and Hut78 cells at each time point.After 48-hour treatment,flow cytometry was conducted to detect cell apoptosis,and Western blot analysis to determine expression of apoptosis-related proteins in these cells.Statistical analysis was carried out by using repeated measures analysis of variance,one-way analysis of variance,and least significant difference (LSD)-t test for multiple comparisons.Results Compared with DMSO,chidamide and matrine alone or in combination could inhibit the proliferation of HH and Hut78cells to different extents (F =15.88,558.26,P < 0.05,< 0.001,respectively).At 48 hours,the apoptosis rate in Hut78 cells was significantly higher in the matrine group (20.98% ± 1.53%),chidamide group (22.44% ± 7.74%) and combination group (44.53% ± 1.85%) than in the control group (8.42% ± 4.23%;LSD-t =4.76,5.31,13.69 respectively,all P < 0.05),as well as in the combination group than in the matrine group and chidamide group (LSD-t =8.93,8.37 respectively,both P < 0.01);no significant differences were observed in the apoptosis rate of HH cells between the matrine group (13.98% ± 3.86%)or chidamide group (13.61% ± 1.62%) and control group (11.44% ± 1.43%,both P > 0.05),while the combination group (20.94% ± 0.64%) showed a significantly higher apoptosis rate compared with the control group,matrine group and chidamide group (LSD-t =7.37,5.40,5.69 respectively,all P < 0.05).In the case of HH cells,the combination group showed significantly higher cleaved caspase-3 expression (all P < 0.05),but significantly lower protein expression of E-cadherin,nuclear factor (NF)-κB,phosphorylated-Bad (p-Bad) and Bcl-2 compared with the other 3 groups (all P < 0.05).In the case of Hut78 cells,the expression of E-cadherin,NF-κB,p-Bad and Bcl-2 was significantly lower in the matrine group,chidamide group and combination group than in the control group (all P < 0.05),while cleaved caspase-3 expression was significantly higher in the chidamide group and combination group than in the control group (both P <0.05).No matter in HH cells or in Hut78 cells,there were no significant differences in Bad protein expression between the 4 groups (all P > 0.05).Conclusion Chidamide in combination with matrine can inhibit proliferation and induce apoptosis of HH and Hut78 cells,likely by regulating the expression of apoptosis-related proteins E-cadherin,NF-κB,p-Bad,Bcl-2 and cleaved caspase-3.
3.Natural orifice transluminal endoscopic surgery:a preliminary fresh cadaver experiment in dogs model
Yongzhi YANG ; Yu JING ; Xinglan HUANG ; Xueping PAN ; Yunshu TANG ; Kuo LUO ; Yu CAO ; Xiaoxue SONG ; Shiyou WU ; Shuang LIANG ; Depei HE
Chongqing Medicine 2014;(24):3204-3205,3209
Objective The optimal access for natural orifice transluminal endoscopic surgery is still uncertain .This study was designed to compare the practicability and maneuverability of transgastric ,transunmbilical ,and transrectal approach in abdominal surgery in a canine model .Methods Three dogs were used in this research .Three approach :trangastric ,transunmbilical and tran-srectal approach were carried out for abdominal exploration ,liver biopsy ,bladder biopsy and an attempted cholecystectomy .The ma-neuverability ,endoscopic image ,performer′s perception ,and spatial orientation were evaluated .Results The maneuverability of trangastric ,and transrectal approach NOTES were better than transunmbilical NOTES .Abdominal exploration ,live biopsy ,and bladder biopsy were completed successfully .The cholecystectomy was failed because of poor exposure and difficulty of separating the around tissure .Conclusion The optimal approach for upper abdomen NOTES is transrectal route .For lower abdomen NOTES , the trangastric approach is superior to other accesses .Further study is needed to develop more flexible and precise equipment for NOTES and to evaluate more feasible access approach .


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