1.To Explore the Mechanism of Icariin Combined with Prednisone on Steroid-Resistant Nephrotic Syndrome Based on Network Pharmacology and Experimental Verification
Juan LYU ; Enlai DAI ; Yunxia ZHANG ; Junyuan BAI ; Xiaowei PU
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(7):2426-2436
Objective Based on network pharmacology,molecular docking and animal experiment,this study explored the mechanism of Icariin combined with Prednisone in the treatment of steroid-resistant nephrotic syndrome(SRNS)in rats.Methods The targets of Icariin active components were determined by TCMSP,GeneCards and CTD databases.The targets of SRNS were screened from GeneCards,OMIM,CTD and DRUGBANK databases,Remove and integrate.Venn graph was drawn to obtain the intersection target.A protein-protein interaction network was constructed using STRING database,and core targets were screened by topological analysis using Cytoscape 3.7.2.DAVID 6.8 was employed for GO term enrichment and KEGG pathway enrichment.And AutoDockTools Vina,PyMOL for molecular docking.SRNS rat model was constructed.The body weight of rats was measured,The 24-hour urinary protein quantity was detected,and the pathological morphology of renal tissue was observed by HE staining.,and the expression of core target proteins was de tected by Western blot.Results Network pharmacologic analysis showed that 139 targets corresponding to Icariin active ingredients were obtained,and 58 common targets were obtained by intersection with 1476 SRNS related targets.In PPI network,the top three degree values were Akt,JUN and IL6.KEGG enrichment analysis showed that Icariin played a therapeutic role in SRNS mainly by PI3K/AKT singnaling pathway.Molecular docking verification showed that Akt,PI3K and autophagy-related protein LC3-II had good binding activity with Icariin.Animal experiment result showed that Icariin combined with Prednisone significantly increased the body weight of SRNS rats(P<0.05),decreased the 24 hour urine protein quantity(P<0.01),and improved the glomerular morphological changes.The results of Western-blotting showed that Icariin combined with Prednisone could significantly improve the abnormal decrease of LC3-II protein and the abnormal increase of Akt and PI3K protein expression in renal tissue of SRNS rats(P<0.01).Conclusion Icariin can play a role in the treatment of SRNS through a multi-target and multi-pathway,regulate the PI3K-Akt signaling pathway,and increase the autophagy activity of kidney tissue in SRNS rats,which may be one of its main mechanisms.
2.Effects of decorin on proliferation, migration and invasion of bladder cancer cells
Ziyi WANG ; Hongjie CHEN ; Ninggang YANG ; Jun ZHANG ; Xiangjun ZHANG ; Xinning YU ; Zhongyi MA ; Enlai DAI
Journal of International Oncology 2021;48(6):335-340
Objective:To investigate the effects of decorin (DCN) on the proliferation, migration and invasion of bladder cancer cells.Methods:Bladder cancer T24 cell line was used as the research object. MTT assay was used to detect the inhibitory effect of DCN at different concentrations (0, 5, 10, 20, 30, 40, 50 mg/L) on T24 cell proliferation at 24, 48, 72 and 96 h. The effects of DCN on T24 cell cycle and apoptosis were analyzed by flow cytometry. MTT assay, Transwell migration and invasion experiments were used to detect the effects of DCN on the adhesion, migration and invasion ability of T24 cells. The effects of DCN on TGF-β1 and P21 protein expression were detected by ELISA and Western blotting.Results:T24 cells were treated with 0, 5, 10, 20, 30, 40 and 50 mg/L DCN at 24, 48, 72 and 96 h, and there were statistically significant diffe-rences in cell proliferation activity ( F=168.64, P<0.001; F=165.81, P<0.001; F=291.02, P<0.001; F=148.93, P<0.001). T24 cells were treated with 0, 5, 10, 20, 30, 40 and 50 mg/L DCN for 72 h, and the cell proliferation activities were (60.71±3.03)%, (40.82±2.09)%, (37.24±1.63)%, (25.65±2.55)%, (23.00±2.67)%, (10.78±1.17)%, (11.04±0.96)%, respectively, and there was a statistically significant difference. At the concentration of 40 mg/L, the proliferation activity reached the lowest level, and the inhibitory effect on cell proliferation was the strongest. At concentrations of 40 and 50 mg/L, the cells in G 1 phase reached the peak value, while the cells in S phase reached the lowest value, and the cells in G 2 phase remained unchanged throughout the treatment process. T24 cells were treated with 0, 5, 10, 20, 30, 40 and 50 mg/L DCN for 72 h, and the apoptosis rates of cells were (12.18±1.17)%, (21.24±1.05)%, (19.80±1.20)%, (26.52±1.40)%, (30.86±1.40)%, (52.99±1.22)%, (43.04±2.16)%, respectively, and there was a statistically significant difference ( F=178.54, P<0.001). The differences between 5, 10, 20, 30, 40, 50 mg/L DCN and 0 mg/L DCN were all statistically significant (all P<0.001). When T24 cells were treated with 0, 40 mg/L DCN for 72 h, the cell adhesion rates were (37.14±1.35)% and (59.86±1.95)%, the numbers of migrated cells were 53.86±3.18 and 12.86±1.35, and there were statistically significant differences ( t=25.25, P<0.001; t=31.36, P<0.001). When DCN was applied to T24 cells for 48 h, the numbers of invasion at 0, 40 mg/L were 235.14±3.44 and 160.86±3.13, and there was a statistically significant difference ( t=2.27, P<0.001). When T24 cells were treated with 0, 5, 10, 20, 30, 40 and 50 mg/L DCN for 72 h, the relative expression levels of TGF-β1 were 85.67±3.35, 45.51±1.19, 49.93±4.15, 47.64±3.53, 46.05±3.18, 25.54±2.25, 33.44±4.05, and there was a statistically significant difference ( F=324.58, P<0.001). Compared with 0 mg/L DCN, 5, 10, 20, 30, 40 and 50 mg/L DCN could significantly inhibited the expression of TGF-β1 (all P<0.001). Compared with 0 mg/L DCN, P21 protein was upregulated 72 h after treatment with 40 mg/L DCN. Conclusion:DCN can inhibit proliferation and induce apoptosis of T24 cells in vitro, and has the effect of anti-metastasis of T24 cells.
3. Role of heterotrimer G protein in podocyte injury
Can LIU ; Enlai DAI ; Zhaoran DING ; Shuwen DUAN ; Xiaohui WANG ; Enlai DAI
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(2):177-187
GPCRs are the largest membrane protein receptor superfamily in the human body, with more than 800 isoforms, and approximately 35% of Food and Drug Administration-approved and marketed drugs currently target GPCRs for the treatment of a wide range of diseases, for heart failure (beta-adrenergic receptors), peptic ulcer (histamine receptors), prostate cancer (gonadotropin receptors), hypertension (adrenergic and angiotensin receptors), pain (opioid receptors), and bronchial asthma (beta2-adrenergic receptors) examples. Although the number of GPCRs is enormous, the signaling proteins downstream of them are limited, heterotrimeric G proteins (GPs) are key proteins that signal GPCRs, translate extracellular stimuli into intracellular responses by coupling to GPCRs and initiate multiple signaling events via downstream cascades. Podocytes are an important component of the glomerular filtration barrier, and their damage is a central event in proteinuria formation and progressive glomerulosclerosis. This article reviews the regulation of GPs, their signaling and their role in podocyte injury to provide a theoretical basis for scientific research and clinical treatment of this disease.