Sulforaphane, an NRF2 agonist, alleviates ferroptosis in acute liver failure by regulating HDAC6 activity / 中西医结合学报

OBJECTIVE@#Acute liver failure (ALF) is characterized by severe liver dysfunction, rapid progression and high mortality and is difficult to treat. Studies have found that sulforaphane (SFN), a nuclear factor E2-related factor 2 (NRF2) agonist, has anti-inflammatory, antioxidant and anticancer effects, and has certain protective effects on neurodegenerative diseases, cancer and liver fibrosis. This paper aimed to explore the protective effect of SFN in ALF and it possible mechanisms of action.@*METHODS@#Lipopolysaccharide and D-galactosamine were used to induce liver injury in vitro and in vivo. NRF2 agonist SFN and histone deacetylase 6 (HDAC6) inhibitor ACY1215 were used to observe the protective effect and possible mechanisms of SFN in ALF, respectively. Cell viability, lactate dehydrogenase (LDH), Fe2+, glutathione (GSH) and malondialdehyde (MDA) were detected. The expression of HDAC6, NRF2, glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4) and solute carrier family 7 member 11 (SLC7A11) were detected by Western blotting and immunofluorescence.@*RESULTS@#Our results show that NRF2 was activated by SFN. LDH, Fe2+, MDA and ACSL4 were downregulated, while GSH, GPX4 and SLC7A11 were upregulated by SFN in vitro and in vivo, indicating the inhibitory effect of SFN on ferroptosis. Additionally, HDAC6 expression was decreased in the SFN group, indicating that SFN could downregulate the expression of HDAC6 in ALF. After using the HDAC6 inhibitor, ACY1215, SFN further reduced HDAC6 expression and inhibited ferroptosis, indicating that SFN may inhibit ferroptosis by regulating HDAC6 activity.@*CONCLUSION@#SFN has a protective effect on ALF, and the mechanism may include reduction of ferroptosis through the regulation of HDAC6. Please cite this article as: Zhang YQ, Shi CX, Zhang DM, Zhang LY, Wang LW, Gong ZJ. Sulforaphane, an NRF2 agonist, alleviates ferroptosis in acute liver failure by regulating HDAC6 activity. J Integr Med. 2023; 21(5): 464-473.

Advanced glycation end products promote differentiation of CD4(+) T helper cells toward pro-inflammatory response / 华中科技大学学报（医学）（英德文版）

This study investigated the effect of advanced glycation end products (AGEs) on differentiation of naïve CD4(+) T cells and the role of the receptor of AGEs (RAGE) and peroxisome proliferator-activated receptors (PPARs) activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin (BSA) with glucose. Human naïve CD4(+) T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin (sh) RNA knock-down experiment, naïve CD4(+) T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-X(TM) 293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4(+) T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T (Treg) cells was determined by a [(3)H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from naïve CD4(+) T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in naïve CD4(+) T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4(+) T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα PPARγ agonist, PGJ2, inhibited the effect of AGEs on naïve CD4(+) T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4(+) T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity.

Advanced glycation end products promote differentiation of CD4(+) T helper cells toward pro-inflammatory response / 华中科技大学学报（医学）（英德文版）

This study investigated the effect of advanced glycation end products (AGEs) on differentiation of naïve CD4(+) T cells and the role of the receptor of AGEs (RAGE) and peroxisome proliferator-activated receptors (PPARs) activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin (BSA) with glucose. Human naïve CD4(+) T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin (sh) RNA knock-down experiment, naïve CD4(+) T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-X(TM) 293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4(+) T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T (Treg) cells was determined by a [(3)H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from naïve CD4(+) T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in naïve CD4(+) T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4(+) T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα PPARγ agonist, PGJ2, inhibited the effect of AGEs on naïve CD4(+) T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4(+) T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity.

Effects of hydroxycamptothecin on TGFb1, a-SMA and collagen I expression in rat hepatic satellite cells / 中华肝脏病杂志

To investigate the molecular mechanism of hydroxycamptothecin (HCPT)-mediated anti-hepatic fibrosis by evaluting its effects on expression of tumor growth factor-beta 1 (TGFb1), alpha-smooth muscle actin (a-SMA) and collagen I (Col I) in hepatic satellite cells (HSCs). Cultured HSCs were treated with different concentrations of HCPT: low-dose group, 0.25 mg/L; middle-dose group, 0.5 mg/L; high-dose group, 0.75 mg/L; and control group, 0 mg/L. Cell proliferation was assessed by the MTT assay. The mRNA expressions of TGFb1, a-SMA and Col I were determined by reverse transcription-polymerase chain reaction. The protein expressions of TGFb1 and a-SMA were detected by Western blot. The content of Col I in the cultured HSCs' supernatant was measured by enzyme-linked immunosorbent assay. The MTT absorbance values of the low-dose group (0.631+/-0.074), middle-dose group (0.469+/- 0.012) and high-dose group (0.204+/- 0.001) were significantly lower than that of the control group (0.793+/-0.098; F = 82.86, P less than 0.01). Compared with the control group, the HCPT-treated groups showed significantly down-regulated gene expressions of TGFb1 (control: 0.716+/-0.064 vs. low: 0.611+/-0.040, middle: 0.510+/-0.014, high: 0.403+/-0.026), a-SMA (control: 0.696+/-0.075 vs. low: 0.579+/-0.037, middle: 0.470+/-0.024, high: 0.299+/-0.017), and Col I (control: 1.019+/-0.056 vs. low: 0.835+/-0.022, middle: 0.696+/-0.055, high: 0.322+/-0.104) (all, P less than 0.01). Meanwhile, HCPT-treated HSCs showed significantly reduced protein expressions of TGFb1 (control: 0.872+/-0.053 vs. low: 0.654+/-0.047, middle: 0.545+/-0.042, high: 0.436+/-0.039) and a-SMA (control: 0.858+/-0.050 vs. low: 0.620+/-0.045, middle: 0.525+/-0.042, high: 0.434+/-0.052) (all, P less than 0.01). The Col I levels secreted by HSCs were significantly lower in the HCPT-treated groups (low: 168.367+/-16.453 ng/ml; middle: 141.284+/-11.731 ng/ml; high: 132.910+/-10.048 ng/ml) than in the control group (188.733 +/-18.299 ng/ml) (all, P less than 0.01). The mechanism of HCPT-mediated anti-hepatic fibrosis may involve down-regulation of TGFb1 expression to inhibit HSC proliferation and activation, as well as reduction of Col I synthesis and secretion.

Expression and subcellular localization of APOBEC3G in peripheral blood mononuclear cells and liver tissues of chronic HBV patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the expression level and intracellular localization of APOBEC3G in peripheral blood mononuclear cells (PBMCs) and liver tissues of chronic HBV patients.</p><p><b>METHODS</b>The expression level and intracellular localization of APOBEC3G in PBMCs and liver tissues were detected using the western blot and confocal laser scanning microscope (CLSM).</p><p><b>RESULTS</b>Western-blot showed that the expression level of APOBEC3G in PBMCs of healthy controls was very low. The relative expression levels of APOBEC3G in PBMC of patients with chronic hepatitis B, chronic severe hepatitis, liver cirrhosis, or liver cancer were 4.12+/-0.21, 4.07+/-0.28, 4.16+/-0.36 or 4.21+/-0.39 respectively, which were higher than that in the healthy controls. However, there was no significant difference in APOBEC3G expression among different chronic HBV patients (q = 0.931, 0.744, 1.675, 1.675, 2.606 or 0.931, respectively, all P values more than 0.05). In addition, there was no significant difference on APOBEC3G in liver tissues between chronic hepatitis B patients and hepatocellular carcinoma patients (4.40+/-0.34 vs 4.34+/-0.43, q = 0.588, P more than 0.05). CLSM indicated that the localization of APOBEC3G protein was in cytoplasm of PBMCs and hepatocytes.</p><p><b>CONCLUSION</b>APOBEC3G is upregulated in the PBMCs of chronic hepatitis B patients.</p>

Human plasmacytoid dendritic cells from patients with chronic hepatitis B induce the generation of CD4+CD25+Treg in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To compare the ability of plasmacytoid dendritic cells (pDCs) from chronic HBV patients and healthy controls to induce the the generation of CD4+CD25+Treg cells.</p><p><b>METHODS</b>The pDCs were isolated from PBMCs of 46 chronic HBV patients, 10 resolved HBV patients and 25 healthy controls by magnetic cell sorting. Purified CD4+CD45RA+ naive T cells were incubated with allogeneic pDCs from chronic HBV infected patients, resolved HBV patients or healthy controls. The cells were stimulated with HBcAg or tetanus toxin. The proportion of CD4+CD25+Treg in CD4+ T cells primed by pDCs was determined by flow cytometry, the expression of Fox p3 mRNA was detected by RT-PCR, IL-10 and TGFb1 expression was quantified using ELISA kits.</p><p><b>RESULTS</b>Compared with pDC isolated from healthy controls and the resolved HBV patients, pDC from chronic HBV patients was more effective in suppression of CD4+ T cells proliferation and interferon production when CD25-depleted PBMCs were stimulated with HBcAg, (7999.36+/-374.74 vs 11 282.56+/-1174.46; 7999.36+/-374.74 vs 12 304.58+/-1462.81, P less than 0.05 ). Depletion of CD4+CD25+ Treg from CD4+ T cells primed by pDC led to the lose of capability to suppress HBV-specific T-cell responses. When CD25- depleted PBMCs were stimulated with purified tetanus toxin, there was no significantly difference in proliferation between CD25-depleted PBMC co-cultured with pDC-primed CD4+ T cells and CD25-depleted PBMC cultured without pDC-primed CD4+ T cells. A higher percentage of CD4+CD25+ Treg was detected within the population of CD4+ T cells primed by pDC from chronic HBV patients compared with healthy controls and resolved HBV patients (5.99%+/-1.85% vs 3.04%+/-0.79%; 5.99%+/-1.85% vs 3.01%+/-1.53%, P less than 0.05). Accordingly, CD25+Treg from pDC-primed CD4+ T cells displayed a higher Fox P3 mRNA level. The IL-10 and TGFb1 could be also detectable in the supernatants of pDC-primed CD4+ T cells.</p><p><b>CONCLUSION</b>pDCs from chronic hepatitis B induce the generation of a higher proportion of CD4+CD25+ Treg compared with pDCs from healthy controls.</p>

A clinical study of adefovir dipivoxil treatment for chronic hepatitis patients with cirrhosis in their decompensation period / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of 96 weeks adefovir dipivoxil (ADV) treatment for chronic hepatitis patients with cirrhosis in their decompensation period.</p><p><b>METHODS</b>Chronic hepatitis patients with cirrhosis in their decompensation period were randomly divided into two groups. An ADV group: patients treated with 10 mg ADV per day; a lamivudine (LMV) group: patients treated with 100mg LMV per day. The course of treatment lasted 96 weeks. Serum levels of ALT, AST, Alb, Tbil, HbeAg, HBV DNA, PCIII, IVC, LN and HA, renal function, Child-Pugh scores, drug adverse reactions and complication during the treatment of the two groups were analyzed.</p><p><b>RESULTS</b>During the two-year treatment, the proportions of patients with a return to normal liver function were similar in both groups. With the treatment prolonged, serum HBV DNA levels of the patients in adefovir dipivoxil group decreased gradually. The HBV DNA level in some lamivudine-treated patients was increased. The sero-conversion rate of HBeAg/HBeAb of the patients in the two groups was increased with the prolongation of the treatment. At 96 weeks, the ratio of emerging virus-resistant strains was lower in the adefovir dipivoxil group than in the lamivudine group. The levels of the serum markers of hepatic fibrosis of the patients in the two groups remained low. Child-Pugh scores of the patients in the two groups were significantly improved. No significant difference in the total incidence of complications between the two groups was noticed. Each of the two groups had a patient with liver-kidney syndrome and other serious complications.</p><p><b>CONCLUSION</b>The efficacy and safety of adefovir dipivoxil and lamivudine treatment for the above patients are similar, but the ratio of emerging virus-resistant strains of the adefovir dipivoxil treatment group is lower than that of lamivudine treatment group.</p>

Study on molecular epidemiology of people infected with human immunodeficiency virus-1 in Hubei province / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the distribution of human immunodeficiency virus-1 (HIV-1) genotypes in Hubei province.</p><p><b>METHODS</b>Epidemiological survey was carried out to HIV-1 carriers who were identified in Hubei province. HIV-1 env V3-V4, gag P17/24 and the first exon of tat region were amplified by nested-polymerase chain reaction(nPCR) .The sequences were determined, and phylogenetic analyses were then performed.</p><p><b>RESULTS</b>4 HIV-1 strains or circulating recombinant forms (CRFs) were identified in Hubei province with subtype B' the predominant which covered 5 kinds of populations including former blood donors, blood receivers, spouses of the infected people, sex workers and their clients, homosexuals, mainly distributed in the areas with many former blood donors. CRF08-BC and CRF01-AE were found distributed in economically more developed cities or southern area of the province, and the major transmission routes was through sexual contact. Only 1 patient, an injecting drug user, was identified having subtype C.</p><p><b>CONCLUSION</b>Subtype B' was the main epidemic subtypes in Hubei province while CRF08-BC, CRF01-AE and subtype C were also circulating in the province, indicating the transmission of the disease might to become more complex.</p>

A clinical study of adefovir dipivoxil treatment for chronic hepatitis patients with cirrhosis in their decompensation period / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of 48 weeks adefovir dipivoxil (ADV) treatment for chronic hepatitis patients with cirrhosis in their decompensation period.</p><p><b>METHODS</b>Sixty-two chronic hepatitis patients with cirrhosis in their decompensation period were randomly put into two groups. An adefovir dipivoxil (ADV) group: 32 patients treated with 10 mg of ADV a day; and a lamivudine (LMV) group: 30 patients treated with 100 mg of LMV a day. The course of treatment lasted 48 weeks. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), Alb, TBil, HBeAg, HBV DNA, PCIII, IVC, LN, and HA, renal function, Child-Pugh scores and drug adverse reactions during the treatment of the two groups were checked, compared and analyzed.</p><p><b>RESULTS</b>The ratios of recovery for liver functions and the negativity rate of HBV DNA, HBeAg, including sero-conversion rate of HBeAg/HBeAb, were increased with prolongation of the treatment period; however, the differences between the two groups were not statistically significant (P > 0.05). Two patients treated with lamivudine suffered from YMDD variation at the 48th week; the ratio of variation was 6.7%. No YMDD variation happened in the ADV group. On the 24th week of the treatment, the levels of the serum markers of hepatic fibrosis declined obviously, compared with those prior to the treatment (P < 0.01). There were no significant statistical differences of those levels between the two groups (P > 0.05). No significant differences of Child-Pugh scores were noticed between the two groups (P > 0.05). No drug related renal function impairment was found during the treatment. Two patients of each group had adverse drug reactions but all were mild.</p><p><b>CONCLUSION</b>The efficacy and safety of adefovir dipivoxil and lamivudine treatment for the above patients were similar, but the ratio of emerging virus-resistant strains was lower in the adefovir dipivoxil treatment group.</p>

Relationship between Shenzhen HBV genotype and precore/core promoter mutation and antiviral effects / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To study the relationship between hepatitis B virus genotyping Shenzhen isolates and HBV precore/core promoter mutation and antiviral effects.</p><p><b>METHODS</b>The HBV genotyping of 165 patients with HBV was carried out with mAbs ELISA. HBV precore/core promoter mutation was detected with gene chip technology in 24 patients with CHB. The relationship between HBV genotyping and interferon, lamivudine effects was analyzed.</p><p><b>RESULTS</b>(1) Out of 165 cases, 106 (64.2%) of type B but 48 (29.1%) of type C were found. Type B accounted for 95.4% in group ASC, and type C for 64.7%in group LC (P<0.05). (2) Precore/core promoter mutation was found in 16 cases (10 of type B, and 6 of type C) out of 24 cases. Out of 16 cases, precore/core promoter mutation (nt1896, 1862) was found in 10 cases (9 cases of type B and 1 case of type C), while basal core promoter mutation (BCP mutation, nt1762,1764) was found in 6 cases (1 case of type B and 5 of type C). (3) Among 27 patients with CHB HBAg (+) treated with interferon, 11 cases of type B but 1 case of type C were tested to be fully responsive to interferon. Among 29 patients with CHB HBAg (+) treated with lamivudine, 15 cases of type B but 3 cases of type C were tested to be continuously responsive to lamivudine.</p><p><b>CONCLUSION</b>(1) HBV genotype popularity in Shenzhen area was classified as type B the first and type C the second. (2) Type C seems more apt to develop BCP mutation and cirrhosis, and to be less responsive to interferon or lamivudine.</p>

The study on quality of life and related factors among patients with chronic hepatitis B in Wuhan area / 中华全科医师杂志

0.05).Conclusion Coverage of medical insurance and effective antiviral therapy for the patients with CHB could affect their QOL.

Effects of perindopril and valsartan on the expression of TGF beta 1 and TGF beta receptor II mRNA, Smad3 and Smad7 in experimental hepatic fibrotic rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic effects of perindopril, an angiotensin-converting enzyme inhibitor, and valsartan, an angiotensin II receptor blocker on TGFbeta1 and TGFbeta receptor II mRNA, Smad3 and Smad7 on rat liver fibrosis.</p><p><b>METHODS</b>60 Wistar rats were randomly divided into four groups (each group, n=15). Group 1 rats were not treated and served as healthy controls. The rats of groups 2,3,and 4 were injected with CCl(4) which induced liver fibrosis. After four weeks, group 3 rats started a treatment of perindopril, and group 4 rats with valsartan. All rats were sacrificed at the eighth week and their blood and livers were collected for analysis. The effects of perindopril and valsartan were evaluated by the levels of transforming growth factor-beta1 (TGFb1), and TGF receptor (TGFb1RII) mRNA in liver tissues by RT-PCR, the expressions and sites of TGFb1, Smad3 and Smad7 in liver tissue by immunohistochemical staining. The liver histopathology was also examined with HE staining, and the hydroxyproline in the liver and serum hyaluronic acid (HA) were examined using biochemsitry and RIA.</p><p><b>RESULTS</b>Compared with the control group, the levels of TGFb1, TGFb1RII mRNA and the expression Smad3 were significantly decreased in the two treated groups, and the expression of Smad7 was also remarkably increased in the livers of rats treated with perindopril or valsartan. The histological changes of fibrosis, the hydroxyproline in the livers and HA were also improved in the treated rats.</p><p><b>CONCLUSION</b>Perindopril and valsartan have a protective effect on liver injury and can inhibit hepatic fibrosis induced by CCl(4) in rats. Their mechanisms may be associated with their effects of down-regulating TGFb1, TGFb1RII mRNA and smad3, and up-regulating Smad7 which then resulted in suppressing the activation of hepatic stellate cells.</p>