A negative feedback loop of H19/miR-675/VDR mediates therapeutic effect of cucurmin in the treatment of glioma.

Curcumin (CUR) shows a remarkable antitumor activity against a wide range of cancers such as glioma, but its underlying mechanism remains elusive. In this study, we aimed to explore the potential role of H19/miR-675/vitamin D receptor (VDR) in the effect of CUR against glioma. Real-time polymerase chain reaction and western-blot analysis were used to study the effect of CUR or 1,25-dihydroxyvitamin D (1,25(OH)2 D3 ) on the expression of H19, miR-675, and VDR. In addition, the effect of H19 on VDR expression was also studied. Furthermore, the expression of H19, miR-675, and VDR between CUR-loaded nanoparticles (NPs) and NP groups was compared, and the interaction among H19, miR-675, and VDR was analyzed by in-silicon and luciferase assays. In a dose-dependent manner, CUR and 1,25(OH)2 D3 both downregulated the expression of H19 and miR-675 but increased the expression of VDR. In addition, H19 evidently reduced the mRNA and protein levels of VDR. Furthermore, VDR was confirmed as a target gene of miR-675, which significantly reduced the expression of VDR. Finally, the administration of CUR evidently decreased tumor volume. CUR-loaded NP group exhibited lower levels of H19 and miR-675, while the NP group showed higher levels of VDR mRNA and protein. In summary, it is the first time that the involvement of a negative feedback loop of H19/miR-675/VDR has been demonstrated in the development of glioma. Therefore, H19 might serve as a new biomarker for the diagnosis and treatment of glioma.

TianJiu therapy for α-naphthyl isothiocyanate-induced intrahepatic cholestasis in rats treated with fresh Ranunculus sceleratus L.

ETHNOPHARMACOLOGICAL RELEVANCE: TianJiu (TJ) therapy, one type of cold moxibustion, applies to specific acupuncture points with herbal patches of hot nature, providing a constant irritant to the skin until the presence of hyperemia and blistering. Traditional and clinical reports suggest that TJ is an effective therapy for the treatment of jaundice with fresh Ranunculus sceleratus L. (RS), in which protoanemonin is one of the main irritant constituents. However, the therapeutic effect of TJ treatment with fresh RS against intrahepatic cholestasis has not been studied in animal experiments. AIM OF THE STUDY: Present study was undertaken to investigate the effect of TJ treatment with fresh RS against intrahepatic cholestasis in rats and provide an experimental basis for the underlying mechanism of TJ therapy. MATERIALS AND METHODS: Male intrahepatic cholestatic Sprague-Dawley rats induced by 2% α-naphthylisothiocyanate (ANIT, 80 mg/kg B.W.) were treated by TJ therapy with fresh RS. The levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), direct bilirubin (DBIL), total bilirubin (TBIL), total bile acid (TBA), hepatic malondialdehyde (MDA) and nitric monoxide (NO), as well as hepatic body ratio, bile flow and hepatic histopathological assay were measured and evaluated to investigate the therapeutic effect of TJ treatment with fresh RS. Phytochemical analysis of fresh and dried RS was performed by gas chromatography-mass spectrometer (GC-MS). RESULTS: After TJ treatment with fresh RS, the abnormally elevated levels of serum AST, ALT, ALP, DBIL, TBIL and TBA, as well as hepatic MDA and NO at 108 h were reduced significantly (versus model group, P < 0.01). The hepatic body ratio, bile flow and hepatic pathological change of cholestatic rats at 108 h in TJ group were restored when compared with those of model group. Thirty-one compounds including lactones, flavonoids and phenolic acids were identified and determined by GC-MS analysis. The content of protoanemonin in fresh RS (9.49%) was about 25-fold higher than that in dried RS (0.38%). CONCLUSIONS: TJ treatment with fresh RS exhibited good therapeutic effect on ANIT-induced intrahepatic cholestasis in rats, which may be due to the attenuated oxidative stress in the liver tissue. It is rational for the ancients to choose fresh RS as TJ herbal patches because of its abundant protoanemonin with the character of irritant. The qualitative and quantitative results of GC-MS analysis provided the chemical basis of TJ therapy with fresh RS, which can be regarded as a simple and efficient method for the treatment of cholestasis hepatitis.

Molecular cloning and transcriptional regulation of an allograft inflammatory factor-1 (AIF-1) in Zhikong scallop Chlamys farreri.

The allograft inflammatory factor-1 (AIF-1) is one of the key factors associated with inflammatory response. In the present study, the full-length cDNA of AIF-1 was identified from Zhikong scallop Chlamys farreri (named as CfAIF-1) by EST (expressed sequence tag) analysis and RACE (rapid-amplification of cDNA ends) approaches. The cDNA of CfAIF-1 consisted of a 5-terminal untranslated region (UTR) of 58 bp, a 3-UTR of 607 bp with a poly (A) tail, and an open reading frame (ORF) of 468 bp encoding a polypeptide of 155 amino acids with the putative molecular mass of 17.8 kDa. There was an EF hand Ca(2+)-binding motif in the deduced amino acid sequence of CfAIF-1 which was conserved in other AIF-1s. CfAIF-1 shared closer phylogenetic relationship with invertebrate counterparts than vertebrate. The mRNA transcripts of CfAIF-1 were dominantly expressed in hepatopancreas, hemocytes and adductor. During scallop ontogenesis, the CfAIF-1 mRNA was expressed at a low level at early developmental stages from eggs to blastula, and then increased significantly from gastruta to late veliger larvae (P<0.05). Moreover, the mRNA expression levels of CfAIF-1 in the hemocytes of adult scallop were significantly up-regulated during 12-48 h after LPS, PGN and poly I:C stimulation (P<0.01), but there was no significant fluctuation detected after glucan stimulation. Furthermore, the challenge of bacteria Vibrio anguillarum remarkably induced the mRNA expression of CfAIF-1 in hemocytes at 6h (P<0.05) and 12h (P<0.01). All these results collectively indicated that CfAIF-1 might be involved in the immune response during the ontogenesis and contribute to the defense against microbe infection in scallops.