11β-Hydroxysteroid Dehydrogenase Type 1 Inhibitor Development by Lentiviral Screening Based on Computational Modeling

Abstract

In this study, rat and human 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) have been cloned by lentiviral transduction and expressed by CHO-K1 cells. The results showed that recombinant plasmids contained R11bhsd1 or H11bhsd1 have been constructed, which is consistent with the gene bank respectively. A clone cell was selected with G418 and cultivated to express 11β-HSD1. 11β-HSD1 catalytic activity of rat and human were 99.5 and 98.7%, respectively, determined by scanning radiometer. And the cloned CHO-K1 cells expressed the protein of 11β-HSD1 in a long-term and stable manner, which makes it suitable for screening 11β-HSD1 inhibitor. The three-dimensional structure of 11β-HSD1 was used for studying the interaction between inhibitor and enzyme by the binding poses predicted by AutoDock and LeDock software. The docking results revealed that compound 8 forms 2 hydrogen bonds with the residues of Gly-216 and Ile-218 in 11β-HSD1, that is to say compound 8 maybe a good 11β-HSD1 inhibitor. Moreover, C57BL/6 mice with R11bHsd1 overexpression had a higher body weight, glucose, total cholesterol, and triglyceride levels compared to the mice treated with an empty viral vector. The results might provide a beneficial foundation for selecting inhibitors of 11β-HSD1 or for researching drug candidate mechanisms.

11β-HSD1 Inhibitor Alleviates Non-Alcoholic Fatty Liver Disease by Activating the AMPK/SIRT1 Signaling Pathway

“…The solution of lentivirus was centrifuged (500× g for 10 min), to remove the cellular debris and purify the lentivirus. The administered virus titer was 10 7 [ 18 ]. The tail veins of 16 mice were injected with lentivirus solution, containing pLVX-EF1α-IRES-ZS green plasmid, and the other half were injected with pLVX-11β-HSD1 green plasmid every 3 days, for 6 consecutive injections of 500 μL.…”

Naringenin Attenuates Non-Alcoholic Fatty Liver Disease by Enhancing Energy Expenditure and Regulating Autophagy via AMPK

“…GetBox Plugin, a plugin of PyMOL 2.4 software, was used to obtain the active site for molecular docking. LeDock 1.0 software (version 1.0, http://lephar.com ) was used for molecular docking ( Liu et al, 2018 ). The binding pocket was set at 3.9, 25.0, 15.9, 31.6, 22.2, and 38.7, and the remaining parameters as default.…”

Bone Marrow Mesenchymal Stem Cell-Derived Nanovesicles Containing H8 Improve Hepatic Glucose and Lipid Metabolism and Exert Ameliorative Effects in Type 2 Diabetes

“…In previous studies, we found that H8 reduced the effect of reducing blood glucose concentration and effectively improved insulin sensitivity in T2D mice, and the effect was better than that of curcumin. 41 , 42 In this study, NVs were used as a sustained-release agent to encapsulate H8, and more biologically targeted nanodrugs (H8-BMSCS-NVS) were prepared by taking advantage of the fact that it takes a long time for molecules to enter and exit the NVs. Our results demonstrated that H8-BMSCs-NVs effectively improved insulin resistance, inhibited hepatic steatosis, and restored impaired liver function in T2D mice.…”