1.High glucose-activated NFAT promotes podocyte apoptosis by down-regulation of PGC-1
Enping CHEN ; Ligen DU ; Yinwei WU ; Cuimei YE ; Quanfang LUO
Chinese Journal of Pathophysiology 2017;33(4):620-626
AIM:To explore whether down-regulation of peroxisome proliferator-activated receptor γ coactivator (PGC)-1α induces podocyte apoptosis and its mechanism.METHODS:The podocytes were cultured under high glucose (HG) condition and the cell apoptosis was analyzed by flow cytometry.The methods of real-time PCR and Western blot were used to analyze the mRNA and protein expression of related molecules in the control, HG-treated or siRNA-treated podocytes.RESULTS:The expression PGC-1α at mRNA and protein levels was significantly decreased in HG-injured podocytes.Down-regulation of PGC-1α expression in vitro by siRNA resulted in podocyte apoptosis.The nuclear protein expression of nuclear factor of activated T-cells (NFAT) was significantly increased in HG injured podocytes, indicating the NFAT activation.Down-regulation of PGC-1α expression also decreased the nuclear protein expression of NFAT.Moreover, silencing of NFAT expression by siRNA significantly abolished PGC-1α deficiency-induced podocyte apoptosis.CONCLUSION: Down-regulation of PGC-1α induces podocyte apoptosis.NFAT mediates PGC-1α deficiency-induced podocyte apoptosis.
2.Regulatory of Jiaweisinisan on expression of hippocampal BDNF, NR1 and dental gyrus neurogenesis in rats with chronic stressed-depression
Can YAN ; Yinwei LIU ; Lili WU ; Penghui ZHU ; Yi PAN
Chinese Pharmacological Bulletin 2016;32(4):569-574
Aim To study the regulatory of Jiaweisini-san on expression of hippocampal BDNF, NR1 and dental gyrus ( DG ) neurogenesis in rats with chronic stressed-depression and its possible mechamisms. Methods Chronic unpredictable mild stress was used to establish the rat model of stressed depression. The expression of BrdU, NeuN, brain -derived neurotro-phic factor ( BDNF ) and N-methyl-D-aspartate recep-tor1 ( NR1 ) in hippocampal dental gyrus were detected by fluorescently labeled immunohistochemical method. In addition, BDNFmRNA was detected by in situ hy-bridization. Results Chronic stress could inhibit the proliferation of neural precursors in hippocampal DG ( P<0. 01 );the expression of BDNF decreased signifi-cantly in DG in model rats ( P <0. 01 ) , while the ex-pression of NR1 increased significantly ( P <0. 01 ) . JWSNS and Fluoxetine hydrochloride significantly en-hanced the amount of new proliferating cells and the number of neurons in unit area of DG ( P<0. 01 ) , in-creased the expression of BDNF ( P <0. 01 ) and de-creased the expression of NR1 in DG(P<0. 01). Con-clusion JWSNS could promote the neuronal prolifera-tion in hippocampal DG of rat with chronic stressed-de-pression,and may exert an effect of promoting the pro-liferation of neurons in hippocampal DG by enhancing the expression of BDNF and decreasing the expression of NR1 .
3.Effects of IL-17A and IFN-γ on the fibrosis of CD34 - orbital fibroblasts in Graves orbitopathy
Yi LU ; Yazhuo HUANG ; Yinwei LI ; Yu WU ; Sijie FANG ; Jing SUN ; Huifang ZHOU
Chinese Journal of Experimental Ophthalmology 2020;38(11):923-928
Objective:To observe the effects of interleukin-17A (IL-17A) and interferon-γ (IFN-γ) on CD34 - orbital fibroblasts (OFs) in patients with Graves ophthalmopathy (GO), and to explore the pathogenesis of GO. Methods:Orbital adipose connective tissue and lacrimal gland tissue of 5 patients with GO were collected during orbital decompression surgery.These tissue was cultured by tissue explant culture method.CD34 - OFs were enriched by immunomagnetic beads after passage and expansion.The cultured cells were divided into transforming growth factor-β (TGF-β) group, TGF-β+ 10 ng/ml IL-17A group, TGF-β+ 100 ng/ml IL-17A group, TGF-β+ 1 ng/ml IFN-γ group and TGF-β+ 5 ng/ml IFN-γ group.The fibrosis of the cells was induced with 5 ng/ml TGF-β and then was treated with different concentrations of IL-17A or IFN-γ according to grouping.The expression of fibronectin, collagen type Ⅰ, α-smooth muscle actin (α-SMA) and tissue metalloproteinase inhibitor-1 (TIMP-1) in CD34 - OFs derived from both orbital adipose connective tissue and lacrimal gland tissue were detected by Western blot.This study protocol was approved by an Ethic Committee of Shanghai Ninth People's Hospital (SH9H-2020-TK195-1) and written informed consent was obtained from each patient. Results:For CD34 - OFs derived from orbital adipose connective tissue, the relative expressions of fibronectin, type Ⅰ collagen, α-SMA and TIMP-1 protein were significantly higher in the TGF-β+ 100 ng/ml IL-17A group than those in the TGF-β group and TGF-β+ 10 ng/ml IL-17A group (all at P<0.05); the relative expressions of fibronectin, type Ⅰ collagen and α-SMA in the cells in the TGF-β+ 5 ng/ml IFN-γ group were significantly lower than those in the TGF-β group and the TGF-β+ 1 ng/ml IFN-γ group (all at P<0.05). For CD34 - OFs derived from lacrimal gland, the relative expressions of fibronectin, type Ⅰ collagen, α-SMA and TIMP-1 protein in the TGF-β+ 100 ng/ml IL-17A group were significantly higher than those in the TGF-β group and the TGF-β+ 10 ng/ml IL-17A group (all at P<0.05); the relative expressions of fibronectin, type Ⅰ collagen and TIMP-1 protein in the TGF-β+ 1 ng/ml IFN-γ group were significantly higher than those in the TGF-β group and TGF-β+ 5 ng/ml IFN-γ group, and the relative expression of α-SMA protein was significantly higher than that in the TGF-β+ 5 ng/ml IFN-γ group (all at P<0.05). Conclusions:High level of IL-17A can promote the fibrosis of TGF-β-induced CD34 - OFs, and high level of IFN-γ inhibits the fibrosis of TGF-β-induced CD34 - OFs.