[Mechanism of total flavonoids of Ziziphora clinopodioides in improving atherosclerosis by regulating PI3K/Akt/mTOR pathway].

The present study observed the regulatory effect of total flavonoids of Ziziphora clinopodioides on autophagy and the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) signaling pathways in ApoE~(-/-) mice and explored the mechanism of total flavonoids of Z. clinopodioides against atherosclerosis(AS). ApoE~(-/-) mice were fed on a high-fat diet for eight weeks to induce an AS model. The model mice were randomly divided into a model group, a positive control group, and low-, medium-and high-dose groups of total flavonoids of Z. clinopodioides, while C57BL/6J mice fed on a common diet were assigned to the blank group. The serum and aorta samples were collected after intragastric administration for 12 weeks, and the serum levels of total cholesterol(TC), triglyceride(TG), low density lipoprotein-cholesterol(LDL-C), and high density lipoprotein-cholesterol(HDL-C) were detected by an automatic biochemical analyzer. The serum expression levels of intercellular adhesion molecule-1(ICAM-1), vascular cell adhesion molecule-1(VCAM-1), matrix metalloproteinase-2(MMP-2), and matrix metalloprotei-nase-9(MMP-9) were detected by enzyme-linked immunosorbent assay(ELISA). Oil red O staining was used to observe the aortic plaque area in mice. Hematoxylin-eosin(HE) staining was used to observe the aortic plaque and pathological changes in mice. The expression of P62 and LC3 in the aorta was detected by the immunofluorescence method. The protein expression of LC3Ⅱ/Ⅰ, Beclin-1, P62, p-PI3K, p-Akt, and p-mTOR in the aorta of mice was detected by Western blot. The results showed that compared with the blank group, the serum levels of TC, TG, LDL-C, ICAM-1, VCAM-1, MMP-2 and MMP-9 in the model group were significantly increased(P<0.01 or P<0.05), the content of HDL-C was decreased(P<0.05), intra-aortic plaque area was enlarged(P<0.01), the expression of LC3 in the aorta was significantly down-regulated, P62 expression was up-regulated(P<0.01 or P<0.05), the expressions of LC3Ⅱ/Ⅰ and Beclin-1 in the aortic lysate were significantly down-regulated, and the expressions of p-PI3K, p-Akt, p-mTOR and P62 were significantly increased(P<0.01). The medium-and high-dose groups of total flavonoids of Z. clinopodioides could reduce the serum levels of TC, TG, LDL-C, ICAM-1, VCAM-1, MMP-2, and MMP-9 in AS model mice(P<0.01 or P<0.05), and increase the content of HDL-C(P<0.01 or P<0.05). The aortic plaque area of mice after middle and high doses of total flavonoids of Z. clinopodioides was significantly reduced(P<0.01), the content of foam cells decrease, and the narrowing of the lumen decreased. The total flavonoids of Z. clinopodioides significantly increased the expression of LC3 in the aorta and the expression of LC3Ⅱ/Ⅰ and Beclin-1 in the lysate, and decreased the expression of P62 in the aorta and the expression of p-PI3K, p-Akt, p-mTOR and P62 in the lysate(P<0.01 or P<0.05). The results showed that the total flavonoids of Z. clinopodioides could improve the content of blood lipids and inflammatory factors, and reduce the generation of foam cells and plaques in aortic tissue, and the mechanism may be related to the regulation of PI3K/Akt/mTOR signaling pathway.

Dietary supplementation with synbiotics improves growth performance, antioxidant status, immune function, and intestinal barrier function in broilers subjected to cyclic heat stress.

This study investigated the effects of synbiotics supplementation on growth performance, antioxidant status, immune function, and intestinal barrier function in broilers subjected to cyclic heat stress. One hundred and forty-four 22-day-old male broilers were randomly assigned to one of three treatment groups of six replicates each for a 21-day study, with eight birds per replicate. Broilers in the control group were reared at a thermoneutral temperature and received a basal diet. Broilers in the other two heat-stressed groups were fed a basal diet supplemented without (heat-stressed group) and with 1.5 g/kg synbiotic (synbiotic group). One and a half gram of the synbiotic consisted with 3 × 109 colony forming units (CFU) Clostridium butyricum, 1.5 × 109 CFU Bacillus licheniformis, 4.5 × 1010 CFU Bacillus subtilis, 600 mg yeast cell wall, and 150 mg xylooligosaccharide. Compared with the control group, heat stress increased rectal temperatures at 28, 35, and 42 days of age, respectively (P < 0.05). Birds subjected to heat stress had reduced weight gain, feed intake, and feed efficiency during 22 to 42 days (P < 0.05). In contrast, supplementation with the synbiotic decreased rectal temperature at 42 days of age and elevated weight gain of heat stress-challenged broilers (P < 0.05). Heat-stressed broilers exhibited a lower superoxide dismutase (SOD) activity in jejunal mucosa and a higher malondialdehyde accumulation in serum, liver and jejunal mucosa (P < 0.05), and the regressive SOD activity was normalized to control level when supplementing synbiotic (P < 0.05). Heat stress increased interleukin-1ß (IL-1ß) and interferon-γ (IFN-γ) levels in serum and IL-1ß content in jejunal mucosa of broilers (P < 0.05). Synbiotic reduced IL-1ß level in serum of broilers subjected to heat stress (P < 0.05). Compared with the control group, elevated serum diamine oxidase activity and reduced jejunal villus height were observed in broilers of the heat-stressed group (P < 0.05), and the values of these two parameters in the synbiotic group were intermediate (P > 0.05). Heat stress upregulated mRNA abundance of IL-1ß and IFN-γ and downregulated gene expression levels of occluding and zonula occluden-1 (ZO-1) in jejunal mucosa of broilers (P < 0.05). The alterations in the mRNA expression levels of jejunal IL-1ß and ZO-1 were reversed by the synbiotic (P > 0.05). In conclusion, dietary synbiotics could improve growth performance, antioxidant capacity, immune function, and intestinal barrier function in heat-stressed broilers.

Effects of graded levels of dietary squalene supplementation on the growth performance, plasma biochemical parameters, antioxidant capacity, and meat quality in broiler chickens.

This study was conducted to evaluate the effects of dietary squalene supplementation on the growth performance, plasma biochemical indices, antioxidant status, and meat quality in broilers. Two hundred and forty 0-day-old male chicks were allocated into 5 groups of 6 replicates and were fed a basal diet supplemented with 0 (Control group), 250, 500, 1,000, or 2,000 mg/kg squalene for 42 d. Dietary squalene supplementation linearly increased weight gain and feed efficiency of broilers during the grower and overall periods (P < 0.05). Squalene linearly decreased 21-d malondialdehyde (MDA) level and 42-d glutathione peroxidase (GSH-Px) activity, and both linearly and quadratically decreased 42-d MDA level in plasma (P < 0.05). In contrast, squalene linearly increased plasma reduced form of glutathione (GSH) level on 21 and 42 d and superoxide dismutase activity on 42 d (P < 0.05). Squalene supplementation linearly decreased 21-d MDA accumulation but linearly increased GSH level on 21 d and 42 d and both linearly and quadratically increased 21-d GSH-Px activity in liver (P < 0.05). Supplementing squalene linearly increased pH value at 48 h and linearly decreased lightness at 48 h and 24-h drip loss of breast muscle (P < 0.05). The lightness at 24 h and cooking loss of breast muscle were both linearly and quadratically reduced by squalene (P < 0.05). Dietary squalene administration linearly decreased MDA accumulation but linearly increased GSH level and GSH-Px activity of breast muscle (P < 0.05). Compared with the control group, aforementioned growth performance, antioxidant-related parameters (except 42-d GSH-Px in plasma and breast and hepatic GSH), and meat quality were improved by squalene when its level was 1,000 and 2,000 mg/kg (P < 0.05), with their results being similar between these 2 groups (P > 0.05). It was concluded that squalene administration especially at a level of 1,000 mg/kg can improve growth performance, antioxidant status, and meat quality in broilers, providing insights into its application as a potential feed additive in broiler production.

The Alcohol Extract of Coreopsis tinctoria Nutt Ameliorates Diabetes and Diabetic Nephropathy in db/db Mice through miR-192/miR-200b and PTEN/AKT and ZEB2/ECM Pathways.

The study aims to investigate the effects of the alcohol extract of Coreopsis tinctoria Nutt (AC) on diabetic nephropathy (DN) mice. A total of 30 db/db (DN) mice were divided into 3 groups, which were treated with AC (300 mg/kg/day), metformin (180 mg/kg/day), or saline by gavage for 10 weeks. Ten db/m mice treated with saline were used as normal control (NC group). Body weight (BW) and fasting blood glucose (FBG), HbA1c, 24 h urinary albumin excretion (UAE), and renal pathological fibrosis were analyzed. Expression of miR-192, miR-200b, and proteins in the PTEN/PI3K/AKT pathway was analyzed by qPCR or western blot. The DN mice had significantly higher BW, FBG, and 24 h UAE, as well as more severe pathological fibrosis when compared with NC. Treatment of AC could decrease BW, FBG, and 24 h UAE and alleviated kidney damage. Compared with the NC group, expressions of miR-192 and miR-200b were increased, whereas their target proteins (ZEB2 and PTEN) were reduced in the kidneys of DN mice, which further modulated the expression of their downstream proteins PI3K p85α, P-AKT, P-smad3, and COL4 α1; these proteins were increased in the kidneys of DN mice. In contrast, AC treatment reversed the expression changes of these proteins. These findings demonstrate that AC may protect the kidneys of DN mice by decreasing miR-192 and miR-200b, which could further regulate their target gene expression and modulate the activity of the PTEN/PI3K/AKT pathway to reduce the degree of renal fibrosis.

Two Traditional Chinese Medicines Curcumae Radix and Curcumae Rhizoma: An Ethnopharmacology, Phytochemistry, and Pharmacology Review.

Curcumae Rhizoma, known as Ezhu (Chinese: ), and Curcumae Radix, known as Yujin (Chinese: ), are different plant parts coming from three same species according to China Pharmacopoeia. Actually, they are used in different ways in TCM clinical treatment. Curcumae Rhizoma is mainly used as antitumor drug, while Curcumae Radix has been used as antidepressant and cholagogue. Curcumae Rhizoma and Curcumae Radix are confused in variety and source, even in clinical trials by some nonprofessional workers. So it is important for us to make them clear. This review is aimed at summarizing the ethnopharmacology, phytochemical, and pharmacological differences between Curcumae Radix and Curcumae Rhizoma by SciFinder, CNKI, and so on, to use them exactly and clearly. Further studies on Curcumae Rhizoma and Curcumae Radix can lead to the development of new drugs and therapeutics for various diseases on the basis of the TCM theory.

R406, an orally available spleen tyrosine kinase inhibitor blocks fc receptor signaling and reduces immune complex-mediated inflammation.

Recent compelling evidence has lead to renewed interest in the role of antibodies and immune complexes in the pathogenesis of several autoimmune disorders, such as rheumatoid arthritis. These immune complexes, consisting of autoantibodies to self-antigens, can mediate inflammatory responses largely through binding and activating the immunoglobulin Fc receptors (FcRs). Using cell-based structure activity relationships with cultured human mast cells, we have identified the small molecule R406 [N4-(2,2-dimethyl-3-oxo-4H-pyrid[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine] as a potent inhibitor of immunoglobulin E (IgE)- and IgG-mediated activation of Fc receptor signaling (EC(50) for degranulation = 56-64 nM). Here we show that the primary target for R406 is the spleen tyrosine kinase (Syk), which plays a key role in the signaling of activating Fc receptors and the B-cell receptor (BCR). R406 inhibited phosphorylation of Syk substrate linker for activation of T cells in mast cells and B-cell linker protein/SLP65 in B cells. R406 bound to the ATP binding pocket of Syk and inhibited its kinase activity as an ATP-competitive inhibitor (K(i) = 30 nM). Furthermore, R406 blocked Syk-dependent FcR-mediated activation of monocytes/macrophages and neutrophils and BCR-mediated activation of B lymphocytes. R406 was selective as assessed using a large panel of Syk-independent cell-based assays representing both specific and general signaling pathways. Consistent with Syk inhibition, oral administration of R406 to mice reduced immune complex-mediated inflammation in a reverse-passive Arthus reaction and two antibody-induced arthritis models. Finally, we report a first-inhuman study showing that R406 is orally bioavailable, achieving exposures capable of inhibiting Syk-dependent IgE-mediated basophil activation. Collectively, the results show R406 potential for modulating Syk activity in human disease.