Hepatic TRPC3 loss contributes to chronic alcohol consumption-induced hepatic steatosis and liver injury in mice

Emerging evidence discloses the involvement of calcium channel protein in the pathological process of liver diseases.Transient receptor potential cation channel subfamily C member 3(TRPC3),a ubiquitously expressed non-selective cation channel protein,controls proliferation,inflammation,and immune response via operating calcium influx in various organs.However,our understanding on the biofunction of hepatic TRPC3 is still limited.The present study aims to clarify the role and potential mechanism(s)of TRPC3 in alcohol-associated liver disease(ALD).We recently found that TRPC3 expression plays an important role in the disease process of ALD.Alcohol exposure led to a significant reduction of hepatic TRPC3 in patients with alcohol-related hepatitis(AH)and ALD models.Antioxidants(N-acetylcysteine and mitoquinone)intervention improved alcohol-induced suppression of TRPC3 via a miR-339-5p-involved mechanism.TRPC3 loss robustly aggravated the alcohol-induced hepatic steatosis and liver injury in mouse liver;this was associated with the suppression of Ca^(2+)/calmodulin-dependent protein kinase kinase 2(CAMKK2)/AMP-activated protein kinase(AMPK)and dysregulation of genes related to lipid metabolism.TRPC3 loss also enhanced hepatic inflammation and early fibrosis-like change in mice.Replenishing hepatic TRPC3 effectively reversed chronic alcohol-induced detrimental alterations in ALD mice.Briefly,chronic alcohol exposure-induced TRPC3 reduction contributes to the pathological development of ALD via suppression of the CAMKK2/AMPK pathway.Oxidative stress-stimulated miR-339-5p upregulation contributes to alcohol-reduced TRPC3.TRPC3 is the requisite and a potential target to defend alcohol consumption-caused ALD.

Identification and Functional Analysis of Serum Specific miRNAs in Recurrent Aphthous Stomatitis Patients with Excess-heat or Yin-deficiency

Background:The treatment of Traditional Chinese Medicine(TCM)in recurrent aphthous stomatitis(RAS)based on syndrome differentiation has won the international acceptance,but the molecular mechanism of excess-heat syndrome and yin-deficiency syndrome of RAS remains unclear.Objective:To clarify specific microRNAs(miRNAs)and their functions in RAS patients with excess-heat or yin-deficiency.Methods:Serum samples were collected from patients meeting the RAS diagnostic criteria of excess-heat or yin-deficiency syndrome and healthy individuals.Core miRNAs were then identified under miRNA microarray anal-yses.Target prediction and bioinformatic analyses were carried out and gene-pathway-networks were visualized to better understand the relationship between different genes and pathways.Results:(1)90 individuals meeting the inclusion criteria were collected in this study.Among them,9 miRNAs were screened out in excess-heat syndrome group(EH group),with 1 upregulated and 8 downregulated.And four miRNAs(hsa-miR-20b-5p,hsa-miR-122-5p,hsa-miR-483-5p and hsa-miR-3197)were validated by real-time PCR method.14 miRNAs were screened out in yin-deficiency syndrome group(YD group)(7 upregulated and 7 downregulated).And hsa-miR-17-5p,hsa-miR-106-5p and hsa-miR-20b-5p were validated.(2)A total of 4,776 target genes were identified in EH group which enriched in GO categories including nervous system development and calcium ion binding and pathway such as PI3K-Akt signaling pathway and Calcium signaling pathway.10,172 target genes were identified in YD group which enriched GO categories included protein binding and positive regulation of transcription from RNA polymerase II promoter and pathway included MAPK signaling pathway and Ras signaling pathway.Conclusion:Hsa-miR-20b-5p in patients with RAS could act as the novel target for the classification of the syndrome.It is upregulated in RAS patients with excess-heat syndrome while downregulated in patients with yin-deficiency syndrome.The PI3K-Akt and MAPK signaling pathways and their related target genes may provide new insights into the molecular mechanisms of RAS with excess-heat syndrome or yin-deficiency syndrome,respectively.

Yunpi Qushi Jiangzhuo Recipe Alleviates Lipid Deposition and Reduces Liver Damage in Mice with Non-alcoholic Fatty Liver Disease

Background:Non-alcoholic fatty liver disease(NAFLD)is a multifactorial liver metabolic disease,which af-fects nearly a quarter of the world’s population.Yunpi Qushi Jiangzhuo Recipe(QSJZR)is a traditional Chi-nese medicine compound,which is composed of Amomum kravanh Pierre ex Gagnep,Coix lacryma-jobi L.var.ma-yuen(Roman.)Stapf,Prunus armeniac a L.var.ansu Maxim,Salvia miltiorrhiza Bge,Nelumbo nucifera Gaertn,Alisma plantago-aquatica Linn,Polyporus umbellatus(Pers.)Fries,Poria cocos(Schw.)Wolf,and Artemisia capillaris Thunb.QSJZR has a certain therapeutic effect on NAFLD patients,but its specific mechanism is not clear.Objective:To investigate the effect of QSJZR on high-fat diet(HFD)-fed NAFLD mice and its mechanism.Methods:Twenty-four SPF female C57BL/6 J mice(21.0±0.5 g)were randomly divided into normal diet(ND)group,HFD group and QSJZR group.ND group was given basal diet,while the other two groups were given HFD.Meanwhile,each mouse in QSJZR group was given 0.2 mL/day(containing 2.27 g crude drug per mL)QSJZR,ND group and HFD group were given the same amount of normal saline for 13 consecutive weeks.Then,the serum was collected for biochemical assay,and the liver was removed for pathological examination and biochemical analysis.Results:Body weight and white fat weight of the HFD-induced NAFLD mice significantly decreased after min-istration with QSJZR,while liver weight had no significant change.QSJZR also significantly reduced liver and serum triglyceride levels,and alleviated hepatocyte lipid deposition by regulating genes and proteins expression related to lipid metabolism,including AMPK,SREBP1C,CPT1A and ACC.In addition,compared with HFD group,liver malondialdehyde(MDA)content was lower in QSJZR group,while glutathione peroxidase(GPx)content was higher.Serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels were lower in QSJZR group than those in HFD group.Treatment with QSJZR significantly alleviated liver injury by increasing BCL2/BAX protein ratio and down-regulating ASK1/JNK pathway.Conclusion:Administration of QSJZR to NAFLD mice once daily for 13 weeks can decrease lipid levels,and alleviate liver damage.These results suggest that QSJZR might be used to treat NAFLD,although more studies need to be conducted for further verification.

A comparison of the biological activities of ethyl acetate fractions from the stems and leaves of Penthorum chinense Pursh

Penthorum chinense Pursh(PCP)is a popular traditional medicinal plant in China,widely used for the treatment of a variety of liver diseases.Although it has been long recognized that the main active elements of PCP are contained in ethyl acetate fraction(EAF),little is known so far in terms of the relative effectiveness of EAF derived from the stems versus leaves of this plant.In the current study,we prepared EAF by reflux extraction and sequential extraction from the stems(SEAF)and leaves(LEAF)of PCP and tested their hepatoprotective efficacies.The extract rates and flavonoid contents of LEAF were higher than those of SEAF.EAFs(>50μg·mL^(–1))prevented lipid accumulation in cells and protected against lipotoxicity injury when the concentration exceeded 25μg·mL^(–1).More than 95%free radicals released by 2,2-diphenyl-1-picrylhydrazyl(DPPH)were eliminated by 25μg·mL^(–1)SEAF and 50μg·mL^(–1)LEAF,respectively.Further,EAFs(25μg·mL^(–1))also showed protective antioxidant effects,with the activity of LEAF being significantly higher than that of SEAF.EAFs(10 mg·mL^(–1))also showed similar unspecific bacteriostatic activity.In comparison with SEAF,LEAF contained more flavonoids and had a higher anti-oxidation capability and for these reasons we suggest it should be better for clinical use.