5.Effect of recombinant human resistin on glucose metabolism by AMPK pathway in HepG2 cells
Zhaofan LUO ; Fangping LI ; Hua CHENG
The Journal of Practical Medicine 2016;32(10):1556-1560
Objective To investigate the effect of rh-resistin on glucose metabolism in HepG2 cells and to elucidate whether the underlying mechanisms are related to AMPK pathway. Methods Cells transfected with control siRNA or AMPKα2 siRNA were cultured in 6-well plates and then treated with 50 ng/mL rh-resistin for 24 hours , while untransfected cells were treated with or without 50 ng/mL rh-resistin on the same conditions , followed by serum-starving in glucose-free DMEM for 3 ~ 5 hours in the continued absence or presence of rh-re-sistin. Then the cells were treated with or without insulin for 2 hours. AMPKα2, G6Pase, PEPCK and Glut2 mRNA expression levels were determined by quantitative RT-PCR. The phosphorylation state of AMPK was deter-mined by Western blotting. Glycogen synthesis was measured by the incorporation of D-[U-14C] glucose to glycogen. Results Rh-resistin suppressed the AMPKα2, Glut2 mRNA expressions, and reduced the phosphory-lation level of AMPK and glycogen synthesis on both basal and insulin-stimulated conditions (P < 0.05), while it accelerated G6Pase and PEPCK mRNA expressions on the same conditions (P < 0.05). The mRNA expression levels of G6Pase , PEPCK , Glut2 and the phosphorylation level of AMPK and glycogen synthesis were signifi-cantly different between the rh-resistin group and the rh-resistin in conjunction with AMPKα2 siRNA-treated group. Conclusion Rh-resistin may affect glucose metabolism in HepG2 cell via AMPK pathway.
6.The effect of the recombinant human resistin on lipid metabolism by AMPK pathway in HepG2 cells
Zhaofan LUO ; Fangping LI ; Hua CHENG
The Journal of Practical Medicine 2017;33(11):1743-1747
Objective To investigate the effect of rh-resistin on lipid metabolism in HepG2 cells and to elucidate its relation to AMPK pathway. Methods We treated the HepG2 cells with 50 ng/ml rh-resistin and 0.5 mmol/L palmitic acid,used siRNA technique to inhibite α2 subunite expression of AMPK in HepG2 cells and quantitative RT-PCR to detect ACC1,ACC2,and HL mRNA expression levels of related lipid metabolism genes. The P-AMPK-Thr172 of AMPK and P-ACC-Ser79 of ACC were determined by Western blotting. The Lipid accumu-lation in cells was determined by images of Laser Scanning Confocal Microscope after Nile red staining. Results Rh-resistin decreased the AMPKα2,HL mRNA expressions and the phosphorylation level of AMPK and ACC in both basal and insulin-stimulated conditions (P < 0.05),had no influence on ACC2 mRNA expressions (P >0.05),while it increased ACC2 mRNA expressions and cytoplasmic lipid droplets in the same conditions (P <0.05). Conclusion Rh-resistin may affect lipid metabolism via AMPK pathway with increase of fatty acid synthe-sis and inhibition of triglyceride catabolism,which leading to lipid accumulation in HepG2 cells.
7.Relationship between vertebral artery incisures and the diseased regions evaluated using transcranial Doppler ultrasound
Yan, LI ; Hua, YANG ; Jing, CHENG
Chinese Journal of Medical Ultrasound (Electronic Edition) 2008;5(2):235-247
Objective To explore the relationship between the incisure changes in the intracranial vertebral artery (VA) segments on transcranial Doppler (TCD) and their diseased regions. Methods Incisure changes in VA intraeranial segments on TCD in 24 cases were found. Digital subtract angiography (DSA), computed tomography angiography (CTA)or coler Doppler flow imaging (CDFI)of carotid arteries were performed to confirm their diseased regions. Results (1)The group with end-systolic and pre-diatolic wide incisures was found in 9 cases: DSA or CTA showed 5 cases with severe stenosis, 2 cases with occlusion and 2 cases with congenital tenuity of the affected vertebral artery proximal part(VA-Pr). Within the above 8 cases, ipsilateral subclavian artery (SubA) was normal, 1 case was complicated with moderate stenosis of ipsilateral SubA,and 1 case was complicated with occlusion of contralateral SubA. Haemodynamics test showed (1)negative; (2)The group with systolic incisures was in 12 cases, and 12 patients were divided into two groups based on the haemodynamics test : ① positive group was in 6 cases. DSA showed that VA-Pr was normal but SubA had impaired with different degrees; ② negative group was in 6 cases. DSA showed VA-Pr was with occlusion in 4 cases and 2 cases was with congenital tenuity; The above 2 cases were complicated with stenosis of ipsilateral SubA and in 4 cases ipsilateral SubA were normal; (3) The group with small vibrated sharp waves on Doppler was in 3 cases, and DSA showed VA-Pr occlusion in 3 cases. In the 3 cases,one case was complicated with occlusion of ipsilateral SubA. Blood stealing was present in cervical muscular branches but not present between vertebral arteries . Ipsilateral SubA were normal in 2 cases. Haemodynamic tests showed negative. Conclusions VA incisures were not only present in the early subclavian steal syndrome (SSS) but also were found in the patients with impaired SubA . However, blood steal pathway is imperfect, and would be found in affected VA-Pr or congenital tenuity. Haemodynamic tests can help identify the above situation. The TCD screening method can be used in the routine diagnosis for SubA and VA from proximal to intracranial segments involvement, but there are limitations to some extent. It needs to combine with DSA and CDFI for diagnosis.
8.MR imaging features with pathologic correlation in adenomyosis
Jinwen HOU ; Hua CHENG ; Chuanfu LI
Chinese Journal of Radiology 2001;0(01):-
Objective To evaluate the usefulness of MRI in the diagnosis of adenomyosis. Methods Sagittal Turbo SE T 1WI, T 2WI, T 1SPIR and T 2SPIR MRI examination were performed on 30 cases with adenomyosis. Results The lesions in 12 cases with diffuse adenomyosis were demonstrated as diffusely thickened junctional zone (JZ) measured 10~35 mm (mean 18 mm) On T 2WI. 6 of them were of homogenous hypointensity. Another six had hyperintensive foci within the hypointensive lesions on T 2WI, and these hyperintensive foci could also be found on T 1WI in 5 lesions. 23 focal lesions (adenomyomas) in 18 cases were oval, irregular or round masses with hypointensity that were the same as the intensity of JZ on T 2WI. They were from 2.0-7.5 cm in diameter (mean 3.9 cm), and all but one showed ill-defined margins. 15 masses hadhyperintensive foci within the hypointensive lesions on T 2WI and 12 of 15 presented hyperintensity on T 1WI. The diffusely thickened JZ and focal masses correspond to the hyperplastic and hypertrophic musculature around the heterotopic endometrial islands. The hyperintensive foci were correlated with the heterotopic endometrial islands. The hyperintensive foci showed only on T 2WI corresponded to the heterotopic endometrial islands without hemorrhage. The hyperintensive foci shown on both T 2WI and T 1WI were the heterotopic endometrial islands with hemorrhage. Conclusion MRI is the modality of choice for the diagnosis of adenomyosis. T 2WI is the most useful sequence. T 2WI combining with T 1WI, T 1SPIR, and T 2SPIR can improve the accuracy in the diagnosis of adenomyosis.