Active components and mechanism of Jinwugutong Capsules in treatment of osteoporosis: a study based on UPLC-Q-Exactive-MS/MS combined with network pharmacology / 中国中药杂志

UPLC-Q-Exactive-MS/MS and network pharmacology were employed to preliminarily study the active components and mechanism of Jinwugutong Capsules in the treatment of osteoporosis. Firstly, UPLC-Q-Exactive-MS/MS was employed to characterize the chemical components of Jinwugutong Capsules, and network pharmacology was employed to establish the "drug-component-target-pathway-disease" network. The key targets and main active components were thus obtained. Secondly, AutoDock was used for the molecular docking between the main active components and key targets. Finally, the animal model of osteoporosis was established, and the effect of Jinwugutong Capsules on the expression of key targets including RAC-alpha serine/threonine-protein kinase(AKT1), albumin(ALB), and tumor necrosis factor-alpha(TNF-α) was determined by enzyme-linked immunosorbent assay(ELISA). A total of 59 chemical components were identified from Jinwugutong Capsules, among which coryfolin, 8-prenylnaringenin, demethoxycurcumin, isobavachin, and genistein may be the main active components of Jinwugutong Capsules in treating osteoporosis. The topological analysis of the protein-protein interaction(PPI) network revealed 10 core targets such as AKT1, ALB, catenin beta 1(CTNNB1), TNF, and epidermal growth factor receptor(EGFR). The Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment showed that Jinwugutong Capsules mainly exerted the therapeutic effect by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT) signaling pathway, neuroactive ligand-receptor interaction, mitogen-activated protein kinase(MAPK) signaling pathway, Rap1 signaling pathway and so on. Molecular docking showed that the main active components of Jinwugutong Capsules well bound to the key targets. ELISA results showed that Jinwugutong Capsules down-regulated the protein levels of AKT1 and TNF-α and up-regulated the protein level of ALB, which preliminarily verified the reliability of network pharmacology. This study indicates that Jinwugutong Capsules may play a role in the treatment of osteoporosis through multiple components, targets, and pathways, which can provide reference for the further research.

Protective Effect of Xiangsha Yuyang Decoction on Oxidative Injury in Rats with Gastric Ulcer Based on p62/Keap1/Nrf2 Signal Pathway / 中国实验方剂学杂志

Objective:To confirm the protective effect of Xiangsha Yuyang decoction on acetic acid-induced gastric ulcer model rats and explore its mechanism， so as to provide experimental basis for clinical drug use. Method:The 60 SPF Wistar rats were randomly divided into 6 groups： group， model group， high， middle and low dose groups of Xiangsha Yuyang decoction and omeprazole control group. The rat model of gastric ulcer was induced by acetic acid. The rats in the high， middle and low dose groups of Xiangsha Yuyang decoction were intragastrically administered at the dose of 28，14，7 g·kg^-1， and with omeprazole at the dose of 4.17 g·kg^-1in normal saline， respectively. The rats in the blank group and model group were intragastrically infused with the same volume of normal saline once a day. After 14 days of continuous treatment， the rats were killed， the blood was collected， the area and inhibition rate of gastric ulcer were measured and calculated， the histopathological sections of gastric mucosa were made and the state of gastric mucosal injury was observed， and the changes of gastric mucosal repair factor， gastric tissue related protein， oxidative stress factor and inflammatory factor in serum were detected by enzyme-linked immunosorbent assay（ELISA）. Detected the expression of p62 Kelch-like epichlorohydrin-related protein 1 （Keap1）， nuclear transcription factor E2 related factor 2 （Nrf2） and heme oxygenase-1 （HO-1） signal pathway-related proteins in gastric mucosa by Western blot. Result:Compared with control group， the gastric mucosa of the model group showed obvious pathological changes and a large number of leukocytes infiltrated. In model group， the ulcer area was significantly increased（<italic>P</italic><0.01）， the contents of mucin mucoprotein 5AC （MUC5AC）， epidermal growth factor （EGF）， superoxide dismutase （SOD） and increased prostaglandin E₂ （PGE₂） were significantly decreased（<italic>P</italic><0.01）， the gastrin （GAS）， 8-hydroxydeoxyguanosine （8-OHdG）， malondialdehyde （MDA）， tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， cyclooxygenase-2 （COX-2） were significantly increased. The expression of HO-1 and Nrf2 protein decreased significantly（<italic>P</italic><0.01）， the content of Keap1 increased significantly （<italic>P</italic><0.05）， and the expression of p62 protein decreased. Compared with model group， the hierarchical structure of cells in Xiangsha Yuyang decoction high dose group and omeprazole group were clearer and regular， middle and low dose groups could also repair gastric mucosa to a certain extent. The high and middle dose groups of Xiangsha Yuyang decoction could significantly reduce the gastric ulcer area of acetic acid-induced gastric ulcer rat model （<italic>P</italic><0.01） and increase the ulcer inhibition rate. It can effectively promote the expression of MUC5AC and EGF in gastric mucosa， decrease the level of GAS（<italic>P</italic><0.05，<italic>P</italic><0.01）， decrease the level of 8-OHdG and MDA， increase the activity of SOD（<italic>P</italic><0.01）， decrease the expression level of TNF-<italic>α</italic> and COX-2， increase the content of PGE₂， and significantly increase the amount of Nrf2 and HO-1 protein in gastric mucosa（<italic>P</italic><0.01）. The high dose group of Xiangsha Yuyang decoction could decrease the protein expression of Keap1（<italic>P</italic><0.05） and increase the expression of p62 protein. Conclusion:Xiangsha Yuyang decoction is effective in the treatment of acetic acid-induced gastric ulcer model rats， which can effectively reduce the ulcer area， increase the ulcer inhibition rate and protect the ulcer tissue. Its mechanism may be related to activating p62/Keap1/Nrf2 signal pathway and regulating the expression of related genes so as to improve inflammatory response and regulate oxidative stress response.

Mechanisms of protection effect of bcl-2 gene transfection on heat-stressed cardiomyocytes / 中国应用生理学杂志

<p><b>AIM</b>To study the mechanisms of protection of bcl-2 gene transfection against heat-stressed cardiomyocytes.</p><p><b>METHODS</b>Cardiomyocytes were isolated and cultured. bcl-2 was transfected into cardiomyocytes with Lipofectamine transfection methods. The cardiomyocytes were stressed by heat. The change of H+ -ATPase synthesis activity of cardiomyocytes mitochondria caused by bcl-2 transfection was measured by chemical radiation method. The changes of Caspase 3 activity of cardiomyocytes caused by bcl-2 transfection was measured by fluorometric analysis.</p><p><b>RESULTS</b>bcl-2 transfection could increase the H+ -ATPase synthesis activity of cardiomyocytes mitochondria under heat stress at 41 degrees C and 43 degrees C and could decrease the Caspase 3 activity of cardiomyocytes under heat stress at 41 degrees C and 43 degrees C.</p><p><b>CONCLUSION</b>The protection effect of bcl-2 transfection on heat-stressed cardiomyocytes may be associated with preserved H+ ATPase synthesis activity of cardiomyocytes mitochondria and the activity of Caspase 3 of cardiomyocytes.</p>