Endoplasmic reticulum stress-induced apoptosis up-regulation of DR5 via a CHOP-dependent mechanism in HSC-T6 cells / 中国临床药理学与治疗学

AIM: To investigate the inhibitory effects of endoplasmic reticulum stress(ERS) and TRAIL on hepatic stellate cells in vitro and how their interaction affect the apoptosis of hepatic stellate cells. METHODS: Take thapsigargin (TG) as the endoplasmic reticulum stress-inducing agents, ursodeoxycholic acid (UDCA) for the endoplasmic reticulum stress inhibitors, SP600125 as a c-Jun N-terminal kinase(JNK) inhibitor, HSC-T6 cells were divided into normal control group, DMSO group, TRAIL group, TG group, UDCA group, siCHOP group and SP600125 group. The apoptosis rate of HSC-T6 cell was detected by flow cytometry. Small interference RNA was applied to silence C/EBP homologous protein(CHOP) gene. The protein expression levels of Caspase-8 were detected by immunohistochemistry method. The ERS marker protein CHOP and TRAIL receptor DR5 expression levels were determined by RT-PCR and Western blot. RESULTS:TG (1 μmol/L, 2 μmol/L, 4 μmol/L, 8 μmol/L, 16 μmol/L) increased cell apoptosis rate of HSC-T6. RT-PCR and Western blot showed that the endoplasmic reticulum stress protein marker CHOP could induce the upregulation of TRAIL receptor DR5 and Caspase-8. Moreover, siCHOP and the JNK inhibitor SP600125 could reduce the expression of DR5 and Caspase-8 in HSC cells. CONCLUSION: These results indicated that CHOP and JNK may be a potential factor regulating DR5 expression, and play an important role in the process of apoptosis of hepatic stellate cells.

Influences of different microbial environment on gut microbiota colonization in early life of mice / 中华围产医学杂志

Objective To investigate the influences of exposure to different environmental microbes on early-life gut microbiota colonization in mice.Methods Male (n=8) and female (n=16) adult specific pathogen free (SPF) BALB/c mice were caged together at a ratio of 2:l.After conception,the mice were divided into four groups according to the environments where the offsprings were reared at three different periods (fetal period,breastfeeding period and childhood).Group A:Offsprings were kept in a SPF environment throughout the study;group B:SPF environment during fetal and breastfeeding periods,and then ordinary environment during childhood;group C:SPF environment during fetal period,and then ordinary environment during breastfeeding period and childhood;group D:ordinary environment all the time.Fecal samples were collected at the end of week 3 and 5.Total bacterial DNA was extracted from each sample and analyzed by high throughput analysis.Kruskal-Wallis and Dunn-Bonferroni test were applied for statistical anaysis.Results 1.At the end of three weeks:(1) Diversity:① Phylum level:There were significant differences in the abundance of Firmicutes,Verrucomicrobia,Proteobacteria and Actinobacteria among the four group (all P＜0.01).Compared with group C and D,group A and B showed significantly decreased abundance of Firmicutes [30.876(23.448-41.218)× 10-2,3.317(1.116-4.641) 10-2 vs 71.936(53.587-86.713)× 10-2,79.105(56.305-82.736)× 10-2],but increased abundance of Verrucomicrobia and Proteobacteria [Verrucomicrobia:17.249(9.748-35.106)× 10-2,58.883(0.017-6.047)× 10-2 vs 0.152(0.066-1.890)× 10-2,0.003(0.000-0.016)× 10-2;Proteobacteria:12.640(0.336-15.070)× 10-2,3.653(3.362-4.5955)× 10-2 vs 0.219(0.134-0.325)× 10-2,0.124(0.116-0.165) × 10-2,all P＜0.05 or 0.01].② Genus level:There were significant differences in the abundance of Lactobacillus,Akkermansia and Bacteroides among the four groups (all P＜0.01).Compared with group C and D,group A and B showed significantly decreased abundance of Lactobacillus [19.283(8.618-31.541)× 10-2,0.339(0.264-22.278) × 10-2 vs 58.414(34.874-71.942)× 10-2,66.007(55.141-76.940)× 10-2],but increased abundance of Akkermansia,Bacteroides and Klebsiella [Akkermansia:17.247(9.748-35.106)× 10-2,58.883(0.017-60.475)× 10-2 vs 0.152(0.066-1.890)× 10-2,0.003(0.000-0.017)× 10-2;Bacteroides:3.978(0.683-25.171)× 10-2,8.216(6.023-9.946)× 10-2 vs 0.141(0.061-0.281)× 10-2,0.568(0.149-1.455)× 10-2;Klebsiella:0.209(0.050-8.888)× 10-2,1.402(0.865-1.692)× 10-2 vs 0.003(0.000-0.039) 10-2,0.000(0.000 0.001)× 10-2,all P＜0.05 or 0.01].(2) Alpha diversity:Significant differences were found in operational taxonomic unit (OTU) and Chaol index (P＜0.05),but not in Shannon index among the four groups (P＞0.05).The OTUs of group A and B were significantly lower than that of group D [246(221-348),257(209-280) vs 387(324-478),P=0.045 and 0.008,respectively].2.At the end of five weeks:(1) Diversity:① Phylum level:There were significant differences in the abundance of Firmicutes,Verrucomicrobia and Proteobacteria among the four groups (P＜0.05 or 0.01).The abundance of Firmicutes in gut microbiota in group A was lower than that in group B,C and D [13.765(64.181-24.238)× 10-2 vs 48.912(37.280-59.466)× 10-2,86.065(50.149-89.856) × 10-2,53.847(31.946-72.936) × 10-2],while that of Verrucomicrobia was higher [58.089(22.459-61.285)× 10-2 vs 0.001(0.000-0.005)× 10-2,0.000(0.000-0.001)× 10-2,0.003(0.000-0.006)× 10-2],all P＜0.05 or 0.01.② Genus level:There were significant differences in the abundance of Lactobacillus and Akkermansia among the four groups (P＜0.01).The abundance of Lactobacillus in gut microbiota in group A was lower than that in group B,C and D[1.755(0.805-8.833)× 10-2 vs 26.391(17.550-37.265)× 10-2,70.688(45.713-77.953) × 10-2,28.675 (15.660-57.224) × 10-2],while that of Akkermansia was higher [58.089(22.460-61.285)× 10-2 vs 0.000(0.000-0.006)× 10-2,0.000(0.000-0.001)× 10-2,0.003(0.000-0.006)× 10-2,all P＜0.05 or 0.01].(2) Alpha diversity:There were significant differences in OTU,Chaol and Shannon index among the four groups (P＜0.05 or 0.01).The OTU of group A was lower than that of group B,C and D [268(241-410) vs 438(380-516),562(533-588),546(473-599)],and the OTU,Chaol and Shannon index of group B were all lower than those of group C and D [OTU:438(380-516) vs 562(533-588),546(473-599);Chaol index:1 033(883-1 181) vs 1 285(1 220-1 338),1 328(1 155-1 516);Shannon index:3.85(3.25-4.50) vs 4.28(3.30-5.11),4.17(3.62-4.38),all P＜0.05 or 0.01].Conclusions Early-life exposure to different environments has an obvious impact on the diversity and composition of intestinal microbiota in mice.The less clean the living environment is,the more diverse the gut microflora will be.Furthermore,the window of opportunity for gut microbiota colonization seems to be related to breastfeeding period.

Influence of mode of delivery on post-neonatal gut microbiota / 中华围产医学杂志

Objective To analyze the influence of mode of delivery on post-neonatal gut microbiota using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technology.Methods From April to August in 2013,thirty healthy urban full-term neonates in Nanjing City were enrolled in the study,including fifteen exclusive breastfed ones (seven born of caesarean section and eight born vaginally) and fifteen mixed feeding ones (eight born of cesarean section and seven born vaginally).Stool specimens were collected on the 28th day after birth and bacterial genomic DNA was extracted and examined by PCR on 16S rDNA V3 variable region.Bacterial community profiles were obtained by DGGE.Diversity and similarity differences of the gut microbial community structures were analyzed.Two independent sample t test or Chi-square tests were used for stastistical analysis.Results (1)Diversity analysis showed that among exclusive breastfeeding infants,the Strip number and Shannon-Weaver Diversity Index of gut microbiota in infants born abdominally were significantly lower than those born vaginally [9.71 ±4.27 vs 15.12±4.19,2.13±0.39 vs 2.61±0.32,both P＜0.05],but the Simpson index of gut microbiota was significantly higher [0.13 ± 0.04 vs 0.08± 0.03,P＜0.05],and no significant difference was shown in Pielou Index (P＞0.05).In the mixed feeding group,the Strip number and Shannon-Weaver Diversity Index of gut microbiota in infants born abdominally were significantly lower than those born vaginally [10.88±3.23 vs 16.29±5.38,2.26±0.37 vs 2.66±0.31,both P＜0.05],the Simpson index was higher,but together with the Pielou Index,neither showed significant difference (both P＞0.05).(2) Similarity analysis found that gut microbiota from neonates born of same mode of delivery mostly gathered together and had much more similar structures.Conclusions In the post-neonatal period,the species and numbers of gut microbiota in infants born abdominally are all behind of those born vaginally.The predominant microbiota in babies born of cesarean section are more prominent,and gut microbiota in vaginal delivered babies are more uniformly distributed.

Regulatory effects of total triterpenoid of Prunella vulgarisL. on activities of ERK and TGF-β1/Smad signaling pathway in protecting hepatic fibrosis in rats / 中国药理学通报

Aim To investigate the protective effects of total triterpenoid from Prunella vulgaris L. ( TTP) on CCl4-induced hepatic fibrosis in rats and its mecha-nism. Methods Rat liver fibrosis was induced by 50% CCl4 twice a week for 12 weeks. From the 5th week, all the therapeutic groups were treated with the TTP(25, 50, 100 mg·kg-1 ) and the colchicine (0. 1 mg· kg-1 ) respectively once a day for 8 weeks. At the end of the twelfth week, the levels of ALT, AST, HA, PCⅢ, CⅣ, MDA, SOD, GSH-Px, Hyp were measured . HE and Masson staining were used to evalu-late the degree of hepatic fibrosis. The mRNA expres-sion ofα-SMA, procollagen I, Smad2, Smad3, Smad7 in liver was detected by RT-PCR, and the p-ERK pro-tein expression was evaluated by Western blot. Results Compared with the model group, TTP(25, 50, 100 mg·kg-1 ) not only reduced serum content of ALT, AST, HA, PCⅢ, CⅣand Hyp, MDA in liver tissue, improved the morphologic changes of hepatic fibrosis, but also increased SOD and GSH-Px activity. Moreo-ver, it decreased the α-SMA, procollagen I, Smad2, Smad3 mRNA expression and increased Smad7 mRNA expression in liver tissues obviously. Furthermore, TTP reduced the protein expression of p-ERK. Conclusions TTP can protect rats from CCl4-induced liver fibro-sis. The mechanism of this process may involve inhibi-ting the expression of p-ERK and interference with TGF-β1/Smad signal transduction pathway.

A Review of Methods for Early Evaluation of Alzheimer's Disease / 生物医学工程学杂志

With the intensified aging problem, the study of age-related diseases is becoming more and more significant. Alzheimer's disease is a kind of dementia, with senile plaques and neurofibrillary tangles as the main pathological features, and has become one of the major diseases that endanger the health of the elderly. This review is concentrated on the research of the early assessment of Alzheimer's disease. The current situation of early diagnosis of the disease is analyzed, and a prospect of the future development of early assessment means of the disease is also made in the paper.