Objective To investigate the effects and mechanisms of receptor-interacting serine-threonine kinase 3 (RIP3) in the formation of cholestatic hepatic injury.Methods The experimental study was conducted.(1) Processing and viability of hepatic stellate cell line HSC-T6:HSC-T6 cells were transfected by RIP3-siRNA and NC-siRNA,respectively.The viabilities of un-transfected,RIP3-siRNA-transfected and NC-siRNA-transfected HSC-T6 cells were respectively measured by cell-counting kit-8 (CCK-8).HSC-T6 cells were treated by 100 μmol/L Glycochenodeoxycholic acid (GCDCA) at 0,2,4,8 and 12 hours,and then were extracted and stored,12-hour cell viability was measured by CCK-8.RIP3 that was treated by 100 μmol/L GCDCA knocked down HSC-T6 cells to establishment RIP3 knockdown HSC-T6 cells (RIP3-KD cells).RIP3-KD cells were cultured for 12 hours,and cell viability was measured.(2) Mice model of bile duct ligation (BDL):40 adult mice were randomly divided into 8 groups,5 mice in each group.Sham group:bravery manager was only separated,without ligation,and bloods of inferior vena cava and liver tissues were extracted at 7 days postoperatively.The BDL-1,-3,-5,-7,-14,-21 and-28 d groups:bloods of inferior vena cava and liver tissues were extracted at 1,3,5,7,14,21 and 28 days postoperatively,respectively.(3) The relative expressions of RIP3,α-SMA and TNF-αmRNA in the cells and liver tissues were detected by quantitative real-time polymerase chain reaction (RT-PCR).(4) The relative expressions of RIP3,α-SMA and TNF-α proteins were detected by Western blot.Measurement data with normal distribution were represented as-x±s.The ANOVA was used for data analysis in different time gradient.Comparisons among groups were analyzed using the ANOVA.Pairwise comparison was done by the t test.Results (1) The HSC-T6 cells viability and expressions of RIP3,α-SMA,TNF-α mRNA and proteins:results of CCK8 test showed that 12-hour viabilities of GCDCA-treated HSC-T6 cells,GCDCA-treated RIP3-KD cells,HSC-T6 cells and RIP3-KD cells were 61.3％ ±0.3％ and 83.2％ ±0.4％ and 98.4％ ±0.7％ and 97.4％ ±0.7％ respectively,showing statistically significant differences in the viabilities among them (F =115.200,P＜ 0.05),and showing no statistically significant difference in the viabilities between HSC-T6 cells and RIP3-KD cells (t =1.283,P＞ 0.05).There were statistically significant differences in the viabilities between HSC-T6 cells and GCDCA-treated HSC-T6 cells or GCDCA-treated RIP3-KD cells (t =17.910,6.604,P＜ 0.05) and between GCDCA-treated HSC-T6 cells and GCDCA-treated RIP3-KD cells (t=7.186,P＜0.05).Results of RT-PCR test showed relative expressions of RIP3 mRNA in un-transfected,RIP3-siRNA-transfected and NC-siRNA-transfected HSC-T6 cells were respectively 0.012 1±0.001 3,0.011 2±0.003 1 and 0.002 8±0.000 5,with a statistically significant difference (F =20.410,P ＜ 0.05).There was no statistically significant difference in relative expressions of RIP3 mRNA between un-transfected and NC-siRNA-transfected HSC-T6 cells (t =0.483,P ＞0.05).The relative expression of RIP3 mRNA in RIP3-siRNA-transfected HSC-T6 cells was significant different from that in un-transfected and NC-siRNA-transfected HSC-T6 cells (t =11.760,4.586,P＜0.05).The relative expressions of RIP3 mRNA,α-SMA mRNA and TNF-α mRNA in GCDCA-treated HSC-T6 cells at 0,2,4,8 and 12 hours were 0.012 1±0.001 3,0.011 2±0.003 1,0.021 2±0.002 2,0.027 8±0.002 1,0.029 8±0.002 3 and 0.571±0.012,0.611±0.024,0.691±0.021,0.711±0.021,0.752±0.031 and 0.873±0.022,0.912± 0.024,1.015±0.031,1.210±0.042,1.471±0.041,respectively,showing an increased trend over time and statistically significant differences (F=70.720,30.050,166.700,P＜0.05).The relative expressions of RIP3 mRNA in HSC-T6 cells and GCDCA-treated HSC-T6 cells were 0.012 1±0.001 3 and 0.029 8±0.002 3,with a statistically significant difference (t=13.970,P＜0.05).Results of Western blot showed that relative expressions of RIP3 protein in un-transfected,RIP3-siRNA-transfected and NC-siRNA-transfected HSC-T6 cells were respectively 0.054 ± 0.012,0.013 ± 0.008 and 0.052± 0.021,with a statistically significant difference (F =7.410,P＜0.05).There was no statistically significant difference in relative expressions of RIP3 protein between un-transfected and NC-siRNA-transfected HSC-T6 cells (t =0.143,P ＞ 0.05),and statistically significant differences were found in relative expressions of RIP3 protein between RIP3-siRNA-transfected HSC-T6 cells and un-transfected or NC-siRNA-transfected HSC-T6 cells (t =4.924,3.006,P＜0.05).The relative expressions of RIP3,α-SMA and TNF-oα proteins in GCDCA-treated HSC-T6 cells at 0,2,4,8 and 12 hours were 0.045±0.024,0.047±0.034,0.062±0.025,0.121±0.015,0.154±0.034 and 0.064±0.031,0.072±0.017,0.097±0.035,0.078±0.031,0.254±0.051 and 0.078±0.025,0.094±0.037,0.129±0.041,0.198±0.011,0.324±0.061,respectively,showing an increased trend over time and statistically significant differences (F =9.658,15.810,20.090,P＜0.05).The relative expressions of RIP3 protein in HSC-T6 cells and GCDCA-treated HSC-T6 cells at 12 hours were 0.045±0.024 and 0.154±0.034,with a statistically significant difference (t =4.536,P＜0.05).(2) Expressions of RIP3,α-SMA and TNF-α mRNA in hepatic tissues of mice in each group:the results of RT-PCR showed that relative expressions of RIP3 mRNA,α-SMA mRNA and TNF-α mRNA in the Sham,BDL-1 d,BDL-3 d,BDL-5 d,BDL-7 d,BDL-14 d,BDL-21 d,BDL-28 d groups were 0.047 3±0.003 1,0.041 2±0.007 8-0.339 7±0.017 1 and 2.948±0.612,2.654± 1.032-8.387±0.910 and 0.563±0.078,0.610±0.113-1.078± 0.289,respectively,with statistically significant differences (F =25.180,27.820,7.425,P＜0.05).The results of western blot showed that relative expressions of RIP3,α-SMA and TNF-α proteins in Sham,BDL-1 d,BDL-3 d,BDL-5 d,BDL-7 d,BDL-14 d,BDL-21 d,BDL-28 d groups were 0.245±0.011,0.228±0.023-1.018±0.052 and 0.424±0.057,0.392±0.041-0.985±0.081 and 0.551 ±0.052,0.588±0.087-0.962±0.074,respectively,with statistically significant differences (F=19.160,94.410,22.750,P＜0.05).Conclusion Cholestasis promotes hepatic injury and fibrosis by inducing TNF-α pathway activation and upregulation RIP3.

Objective:To compare the aortic remodeling of the Fabulous stent system and standard thoracic aortic endovascular repair (TEVAR) on distal aorta type B aortic dissection (TBAD). Methods:The prospective data collected between Dec 2017 and Oct 2019 from 134 patients with type B aortic dissection (TBAD) who underwent treatment with the "Fabulous" stent system, and retrospective data from 159 TBAD patients receiving standard TEVAR from corresponding multicenter. By using propensity score matching analysis, we compared the prognosis and aortic remodeling outcomes in patients undergoing Fabulous and standard TEVAR treatments during a 1-year postoperative follow-up.Results:In this study, 62 patients in Fabulous group and 62 patients in standard TEVAR were included.There were no significant statistical differences in baseline characteristics between the two groups. In terms of aortic remodeling in bare stent region, Fabulous group had better change trends of diameter of true lumen [10.6 (4.4, 14.5) mm vs. 4.7 (0.9, 10.7) mm, P=0.001] and false lumen [-24.2 (-30.5, -4.9) mm vs. 0.7 (-11.8, 2.3) mm, P<0.001] than those in the standard TEVAR group. The rate of complete false lumen thrombosis was also higher in the Fabulous group (62.9% vs. 37.1%, P=0.042). Conclusion:The Fabulous stent system, when compared to standard TEVAR surgery, demonstrates good aortic remodeling outcomes in the distal aorta.