Journal: Pharmaceuticals

Article Title: Activation of AMPK/mTOR-Driven Autophagy and Suppression of the HMGB1/TLR4 Pathway with Pentoxifylline Attenuates Doxorubicin-Induced Hepatic Injury in Rats

doi: 10.3390/ph17060681

Figure Lengend Snippet: Pentoxifylline stimulated hepatic adenosine-monophosphate-activated protein kinase/mechanistic target of rapamycin (AMPK/mTOR) pathway in doxorubicin-intoxicated rats. Pentoxifylline (PTX; 100 mg/kg) was orally received daily, while doxorubicin (DOX; 18 mg/kg total cumulative dose; 3 mg/kg dose weekly) was received by the intraperitoneal route. The treatments were administered for 6 weeks. Herein, the protein expression of hepatic p-AMPK(Ser487), total AMPK, p-mTOR(Ser2448), and total mTOR was determined by ELISA. In the hepatic tissue of DOX-intoxicated rats, PTX resulted in an elevated p-AMPK(Ser487)/AMPK ratio ( A ) and diminished p-mTOR(Ser2448)/mTOR ratio ( B ). For n = 6, the values were shown as mean ± S.E.M. * p < 0.05, *** p < 0.001, or **** p < 0.0001 vs. control; # p < 0.05, ## p < 0.01 vs. DOX group. PTX, pentoxifylline; DOX, doxorubicin.

Article Snippet: Moreover, hepatic p-mTOR(Ser2448) and total mTOR levels were determined by ELISA kits supplied by Cell Signaling Technology, Danvers, MA, USA, Cat. No. 7976C and Cat. No. 7974C, respectively).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Pharmaceuticals

Article Title: Neuroprotective Impact of Linagliptin against Cadmium-Induced Cognitive Impairment and Neuropathological Aberrations: Targeting SIRT1/Nrf2 Axis, Apoptosis, and Autophagy

doi: 10.3390/ph16081065

Figure Lengend Snippet: Linagliptin stimulates hippocampal AMPK/mTOR pathway in cadmium-intoxicated rats. Herein, stimulation of AMPK/mTOR pathway by linagliptin in cadmium-intoxicated animals was reflected by the elevation in p-AMPK/AMPK ratio ( B ), and reduction in p-mTOR/mTOR ratio ( D ). Of note, the individual O.D. of p-AMPK(Ser487) and total AMPK are shown in ( A ), whereas the individual O.D. of p-mTOR(Ser2448) and total mTOR are shown in ( C ). For each experimental group, the mean optical density of either p-AMPK or p-mTOR was divided by the corresponding mean optical density of the total AMPK or total mTOR, respectively. This was followed by setting the mean control value to 1. N = 6 in each group (mean ± standard error of the mean). A p -value of less than 0.05 was significant. * p < 0.05, ** p < 0.01, or **** p < 0.0001, compared to control; # p < 0.05, or ## p < 0.01, compared to cadmium (Tukey’s test for multi-comparisons and one-way ANOVA). AMPK, 5′adenosine-monophosphate-activated protein kinase/ mammalian target of rapamycin; Cd, cadmium chloride; LIN, linagliptin; mTOR, mammalian target of rapamycin.

Article Snippet: Likewise, ELISA kits from Cell Signaling were used to quantify phosphorylated and total forms of mTOR (Cat. # 7976C for p-mTOR[Ser2448] and Cat. # 7974C for pan mTOR, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Control

Journal: Molecules

Article Title: The Preventive Effect of Specific Collagen Peptides against Dexamethasone-Induced Muscle Atrophy in Mice

doi: 10.3390/molecules28041950

Figure Lengend Snippet: Effect of CP on myokine alterations. ( A – D ) The mRNA expression of myostatin ( A ), TGF-β ( B ), BDNF ( C ), and FNDC5 ( D ) in the muscles was assessed using real-time PCR. ( E , F ) Total Smad2/3, phospho-Smad2, total-AKT, phospho-AKT, and GAPDH levels in the muscles were assessed by immunoblotting. Total proteins were stained by Ponceau S. ImageJ was used to measure and quantify the band intensity. Arrow in ( E ) indicates p-Smad 2, and this band was used to quantify the p-Smad2 level. The p-Smad2 and p-AKT levels were normalized by Ponceau S staining. ( G ) Total and phospho-mTOR levels were analyzed by ELISA. The p-mTOR levels were normalized against the total mTOR levels. Data are presented as the means ± SD of values obtained from seven biological replicates. # p < 0.05, ## p < 0.01, DEX vs. control. # p < 0.05, ## p < 0.01, DEX + CP (0.25 and 0.5) vs. control. * p < 0.05, ** p < 0.01, DEX + CP (0.25 and 0.5) vs. DEX; CP, collagen peptides; DEX, dexamethasone.

Article Snippet: To quantify the activity of mTOR and p-mTOR, Pathscan sandwich ELISA kits (mTOR: CST#7974, p-mTOR: CST#7976) were used.

Techniques: Expressing, Muscles, Real-time Polymerase Chain Reaction, Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Control

Journal: Molecules

Article Title: The Preventive Effect of Specific Collagen Peptides against Dexamethasone-Induced Muscle Atrophy in Mice

doi: 10.3390/molecules28041950

Figure Lengend Snippet: Anti-inflammatory activity of CP in muscle tissue. ( A ) Evans blue staining of the calf muscle. The blue region in the representative images indicates damaged muscle fibers. The Evans blue-positive area was quantified and is presented as a relative percentage of the control below the representative photos. This experiment was performed with three mice per group. ( B , C ) The mRNA expression of TNF-α and IL-1β in the muscles was assessed using real-time PCR. Data are presented as the means ± SD of values obtained from seven biological replicates. ( D ) The IL-1β levels in serum were examined using ELISA. Data are presented as the means ± SD of values obtained from three biological replicates. # p < 0.05, ## p < 0.01, DEX vs. control. # p < 0.05, ## p < 0.01, DEX + CP (0.25 and 0.5) vs. control. * p < 0.05, ** p < 0.01, DEX + CP (0.25 and 0.5) vs. DEX; CP, collagen peptides; DEX, dexamethasone.

Article Snippet: To quantify the activity of mTOR and p-mTOR, Pathscan sandwich ELISA kits (mTOR: CST#7974, p-mTOR: CST#7976) were used.

Techniques: Activity Assay, Staining, Control, Expressing, Muscles, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay