Shenshuai Yingyang Jiaonang ameliorates chronic kidney disease-associated muscle atrophy in rats by inhibiting ferroptosis mediated by the HIF-1α/SLC7A11 pathway.

Objective: Shenshuai Yingyang Jiaonang (SSYYJN), a traditional Chinese medicine formula, can ameliorate muscle atrophy associated with chronic kidney disease (CKD). However, its mechanisms of action remain unclear. This study is to investigate the molecular mechanisms involved in the effects of SSYYJN in ameliorating muscle atrophy associated with CKD in rats. Methods: The chemical compounds of SSYYJN were identified by UPLC-Q-Orbitrap HRMS. Considering the dose-response relationship of the identified compounds, male SD rats were randomly divided into Sham, Model, SSYYJN, and α-Keto Acid (KA) groups. Subsequently, we assessed the therapeutic and anti-ferroptotic effects of SSYYJN. Network pharmacology studies were used to predict the molecular mechanism of SSYYJN on ferroptosis and were further verified for accuracy. Results: A total of 42 active compounds were identified from SSYYJN. SSYYJN alleviated muscle atrophy caused by CKD, as evidenced by changes in body weight, serum biochemical indices, mass and histopathology of the skeletal muscle, and the levels of MuRF1. SSYYJN reduced the levels of iron, MDA, and ROS, increased the levels of GSH, NAPDH, and Gpx4. Network pharmacology analysis indicated that SSYYJN exerted anti-ferroptotic effects that were closely related to the HIF-1α signaling pathway. Molecular protein and genetic test results showed that SSYYJN increased HIF-1α protein and increased SLC7A11. Conclusions: SSYYJN attenuates muscle atrophy in CKD by inhibiting ferroptosis through the activation of the HIF-1α/SLC7A11 pathway and might be a promising traditional Chinese medicine for muscle atrophy in CKD.

Salidroside alleviates cognitive impairment by inhibiting ferroptosis via activation of the Nrf2/GPX4 axis in SAMP8 mice.

BACKGROUND: Alzheimer's disease (AD) is a neurogenerative disease and remains no effective method for stopping its progress. Ferroptosis and adaptive immunity have been proven to contribute to AD pathogenesis. Salidroside exhibits neuroprotective and immunomodulatory effects. However, the underlying mechanisms linking salidroside, ferroptosis, and adaptive immunity in AD remain uncertain. PURPOSE: The objective of this study is to explore the neuroprotective effects and the potential molecular mechanisms of salidroside against neuronal ferroptosis and CD8+ T cell infiltration in senescence-accelerated mouse prone 8 (SAMP8) mice. STUDY DESIGN AND METHODS: SAMP8 mice were employed as an AD model and were treated with salidroside for 12 weeks. Behavioral tests, immunohistochemistry, HE and Nissl staining, immunofluorescence, transmission electron microscopy, quantitative proteomics, bioinformatic analysis, flow cytometry, iron staining, western blotting, and molecular docking were performed. RESULTS: Treatment with salidroside dose-dependently attenuated cognitive impairment, reduced the accumulation of Aß plaques and restored neuronal damage. Salidroside also suppressed the infiltration of CD8+T cells, oxidative stress, and inflammatory cytokines, and improved mitochondrial metabolism, iron metabolism, lipid metabolism, and redox in the SAMP8 mice brain. The administration of salidroside decreased iron deposition, reduced TFR1, and ACSL4 protein expression, upregulated SLC7A11, and GPX4 protein expression, and promoted the Nrf2/GPX4 axis activation. CONCLUSION: In conclusion, neuronal ferroptosis and CD8+T cells are involved in the process of cognitive impairment in SAMP8 mice. Salidroside alleviates cognitive impairment and inhibits neuronal ferroptosis. The underlying mechanisms may involve the Nrf2/GPX4 axis activation and reduction in CD8+T cells infiltration. This study provides some evidence for the roles of salidroside in adaptive immunity and neuronal ferroptosis in SAMP8 mice.

The Combination of Salidroside and Hedysari Radix Polysaccharide Inhibits Mitochondrial Damage and Apoptosis via the PKC/ERK Pathway.

Background: Beta-amyloid (Aß) peptide is a widely recognized pathological marker of Alzheimer's disease (AD). Salidroside and Hedysari Radix polysaccharide (HRP) were extracted from Chinese herb medicine Rhodiola rosea L and Hedysarum polybotrys Hand-Mazz, respectively. The neuroprotective effects and mechanisms of the combination of salidroside and Hedysari Radix polysaccharide (CSH) against Aß 25-35 induced neurotoxicity remain unclear. Objective: This study aims to investigate the neuroprotective effects and pharmacological mechanisms of CSH on Aß 25-35-induced HT22 cells. Materials and Methods: HT22 cells were pretreated with various concentrations of salidroside or HRP for 24 h, followed by exposed to 20 µm Aß 25-35 in the presence of salidroside or RHP for another 24 h. In a CSH protective assay, HT22 cells were pretreated with 40 µm salidroside and 20 µg/mL HRP for 24 h. The cell viability assay, cell morphology observation, determination of mitochondrial membrane potential (MMP), reactive oxygen species (ROS), and cell apoptosis rate were performed. The mRNA expression of protein kinase C-beta (PKCß), Bax, and Bcl-2 were measured by qRT-PCR. The protein expression levels of cleaved caspase-3, Cyt-C, PKCß, phospho-ERK1/2, Bax, and Bcl-2 were measured by Western blot. Results: CSH treatment increased cell viability, MMP, and decreased ROS generation in Aß 25-35-induced HT22 cells. PKCß and Bcl-2 mRNA expression were elevated by CSH while Bax was decreased. CSH increased the protein expression levels of PKCß, Bcl-2, and phospho-ERK1/2, and decreased those of Bax, Cyt-C, and cleaved caspase-3. Conclusions: CSH treatment have protective effects against Aß 25-35-induced cytotoxicity through decreasing ROS levels, increasing MMP, inhibiting early apoptosis, and regulating PKC/ERK pathway in HT22 cells. CSH may be a potential therapeutic agent for treating or preventing neurodegenerative diseases.

Chemical profiling of Houttuynia cordata Thunb. by UPLC-Q-TOF-MS and analysis of its antioxidant activity in C2C12 cells.

Houttuynia cordata Thunb. ("Yu-Xing-Cao"), a traditional Chinese medicinal herb, has long been used to treat various diseases. However, detailed information regarding the chemical constituents of H. cordata aqueous extract is lacking, and the molecular basis of its beneficial effects on muscle is unknown. To investigate these points, in this study, we used ultra-performance liquid chromatography coupled with quadrupole-time-of-flight-mass spectrometry (UPLC-Q-TOF-MS) in positive and negative ion modes to profile and identify the major constituents of H. cordata water extract. A total of 63 peaks were identified based on mass and fragmentation characteristics, including 29 organic acids and their glycosides, 17 flavonoids, 7 volatiles, 4 pyrimidine and purine derivatives, 2 alkaloids, 2 amino acids, 1 isovanillin, and 1 coumarin. The total flavonoid and polyphenol contents of the extract were 4.77 and 139.15 mg/mL, respectively, by ultraviolet spectrophotometry. The cytoprotective activity of H. cordata aqueous extract was evaluated using C2C12 cells treated with tumor necrosis factor (TNF)-α to induce oxidative challenge. The TNF-α induced decrease in cell viability was reversed by treatment for 48 h with the extract; moreover, superoxide dismutase activity was increased while reactive oxygen species level was decreased. These results provide molecular-level evidence for the antioxidant effect of H. cordata extract and highlight its therapeutic potential for the treatment of muscle injury or diseases caused by oxidative stress.

High-dose vitamin D3 supplementation ameliorates renal fibrosis by vitamin D receptor activation and inhibiting TGF-ß1/Smad3 signaling pathway in 5/6 nephrectomized rats.

Renal fibrosis is the pathological consequence of progressive chronic kidney disease. Although it has been reported that vitamin D3 exerts antifibrotic effects, the underlying mechanisms remain unclear. This study is aimed at investigating the effects and molecular mechanisms in high-dose vitamin D3 treatment on renal fibrosis. A model of chronic kidney disease was established by 5/6 nephrectomy in rats characterised by high levels of serum creatine, urea nitrogen, and urinary protein. Serum 25-dihydroxyvitamin D3, calcium and parathormone levels were measured to evaluate vitamin D levels. Hematoxylin and eosin, periodic acid Schiff and Mallory's Trichrome staining were used to evaluate histopathological changes in rats. Moreover, the expression of vimentin, collagen I, α-smooth muscle actin and E-cadherin were analyzed at molecular and histopathological levels. Our results showed that exposure to vitamin D3 decreased the levels of serum creatine, urea nitrogen and urine protein and restored the homeostasis of calcium and parathormone. Vitamin D3 also downregulated the expression of vimentin, collagen I and α-smooth muscle actin and attenuated renal fibrosis and epithelial to mesenchymal transition in the kidney. Importantly, vitamin D3 treatment increased the expression of the vitamin D receptor and inhibited Transforming growth factor-ß1 (TGF-ß1)/Smad3 signaling pathway in rats kidneys with chronic kidney disease. Mechanistically, the upregulation of TGF-ß1 and phosphorylation of Smad3 induced by vitamin D3 was reversed by activation of the vitamin D receptor. Our findings indicated that vitamin D3 is a potential antifibrotic drug in chronic kidney disease via the vitmin D receptor and inhibiting TGF-ß1/Smad3 signaling pathway.

Atractylenolide III Attenuates Muscle Wasting in Chronic Kidney Disease via the Oxidative Stress-Mediated PI3K/AKT/mTOR Pathway.

Oxidative stress contributes to muscle wasting in advanced chronic kidney disease (CKD) patients. Atractylenolide III (ATL-III), the major active constituent of Atractylodes rhizome, has been previously reported to function as an antioxidant. This study is aimed at investigating whether ATL-III has protective effects against CKD-induced muscle wasting by alleviating oxidative stress. The results showed that the levels of serum creatinine (SCr), blood urea nitrogen (BUN), and urinary protein significantly decreased in the ATL-III treatment group compared with the 5/6 nephrectomy (5/6 Nx) model group but were higher than those in the sham operation group. Skeletal muscle weight was increased, while inflammation was alleviated in the ATL-III administration group compared with the 5/6 Nx model group. ATL-III-treated rats also showed reduced dilation of the mitochondria, increased CAT, GSH-Px, and SOD activity, and decreased levels of MDA both in skeletal muscles and serum compared with 5/6 Nx model rats, suggesting that ATL-III alleviated mitochondrial damage and increased the activity of antioxidant enzymes, thus reducing the production of ROS. Furthermore, accumulated autophagosomes (APs) and autolysosomes (ALs) were reduced in the gastrocnemius (Gastroc) muscles of ATL-III-treated rats under transmission electron microscopy (TEM) together with the downregulation of LC3-II and upregulation of p62 according to Western blotting. This evidence indicated that ATL-III improved skeletal muscle atrophy and alleviated oxidative stress and autophagy in CKD rats. Furthermore, ATL-III could also increase the protein levels of p-PI3K, p-AKT, and p-mTOR in skeletal muscles in CKD rats. To further reveal the relevant mechanism, the oxidative stress-mediated PI3K/AKT/mTOR pathway was assessed, which showed that a reduced expression of p-PI3K, p-AKT, and p-mTOR in C2C12 myoblast atrophy induced by TNF-α could be upregulated by ATL-III; however, after the overexpression of Nox2 to increase ROS production, the attenuated effect was reversed. Our findings indicated that ATL-III is a potentially protective drug against muscle wasting via activation of the oxidative stress-mediated PI3K/AKT/mTOR pathway.

[Phosphorus Fractions and Release Risk in Surface Sediments of an Agricultural Headwater Stream System in Hefei Suburban, China].

A typical water system of agricultural headwater stream in Chaohu Lake basin was selected as the study area, and 17, 16, 14 and 13 surface sediments were collected from the four styles of stream, respectively, including ponds, branches, main channel and mainstream deep pools, in October 2014 (in autumn) and April 2015 (in spring). The forms and space-time variations of phosphorus in the sediments were analyzed. Clustering and variance analysis were conducted on the phosphorus forms data from the four styles of stream by means of multivariate statistical analyses. We quantified the phosphorus release risk (PSI) and identified the main impact factors of PSI via calculating the phosphorus sorption index (PSI) and the correlation analysis. The results showed that: (1) The contents of TP in the surface sediments ranged from 137.517 to 1709.229 mg x kg(-1) with an average value of 532. 245 mg x kg(-1), and the order of the average contents of phosphorus forms was IP (350.347 mg x kg(-1)) > OP (167.333 mg x kg(-1)) > Fe/Al-P ( 78. 869 mg x kg(-1)) > Ca-P (56.343 mg x kg(-1)) > Ex-P (6.609 mg x kg(-1)); (2) The contents of phosphorus forms had the same trend in all the four stream styles, which was deep pool > main channel > branch > pond; (3) In autumn, the deep pool and main channel were clustered into one class, while the pond and branch were clustered into the other class. In spring, branch, main channel and deep pool were clustered into the same class; (4) Variance analysis showed that the differences among the four stream styles were larger in autumn than in spring; (5) The PSI of the surface sediments ranged between 24.49 and 69.94 (mg x L(-1) x (100 g x micromol)(-1). The PSI in spring was lower than that in spring, indicating that phosphorus release risk of surface sediment was higher in spring than in autumn. (6) PSI had a significant negative correlation with Ex-P, IP and pH.

[Soil Phosphorus Forms and Leaching Risk in a Typically Agricultural Catchment of Hefei Suburban].

To investigate the soil phosphorus forms and leaching risk in a typically agricultural catchment of Ershibu River in Hefei Suburban, Chaohu Lake basin, 132 surface soil samples were collected from the catchment area. The spatial distribution of total phosphorus (TP) and bio-available phosphorus (Bio-P), and the spatial variability of soil available phosphorus (Olsen-P) and easy desorption phosphorus (CaCl2-P) were analyzed using the Kriging technology of AreGIS after speciation analysis of soil phosphorus. Moreover, the enrichment level of soil phosphorus was studied, and the phosphorus leaching risk was evaluated through determining the leaching threshold value of soil phosphorus. The results showed that the samples with high contents of TP and Bio-P mainly located in the upstream of the left tributary and on the right side of local area where two tributaries converged. The enrichment rates of soil phosphorus forms were arranged as follows: Ca-P (15.01) > OP (4.16) > TP (3. 42) > IP (2.94) > Ex-P (2.76) > Fe/Al-P (2.43) > Olsen-P (2.34). The critical value of Olsen-P leaching was 18.388 mg x kg(-1), and the leaching samples with values higher than the threshold value accounted for 16.6% of total samples. Generally, the high-risk areas mainly occurred in the upstream of the left tributary, the middle of the right tributary and the local area of the downstream of the area where two tributaries converged.

[Study on chemical constituents from petroleum ether-soluble parts of cones of Platycladus orientalis].

OBJECTIVE: To study chemical constituents from petroleum ether-soluble parts of cones of Platycladus orientalis. METHODS: The compound were isolaeed by repeated column chromatography of silica gel and Sephadex LH-20. The structures were elucidated by physicochemical properties and spectrum analysis. RESULTS: Seven constituents were isolated and identified as sandaracopimaric acid (1), 6alpha-hydroxy sandaracopimaric acid (2), ent-isopimara-8 (14), 15-dien-3beta, 19-diol (3), ent-isopimara-8 (9), 15-dien-3beta-ol (4), ent-isopimara-8 (14),15- dien-3beta-ol (5), isocupressic acid (6) and 15-acetylisocupressic acid (7). CONCLUSION: Compounds 2 and 7 are isolated from this genus for the first time.