Protective effect of Peucedanum praeruptorum Dunn extract on oxidative damage of LLC­PK1 cells induced by H2O2.

Peucedanum praeruptorum Dunn extract (PPDE) is a well-known treatment used in traditional Chinese medicines, where it is most commonly used to treat coughs and symptoms such as headaches and fever. In the present study, the antioxidant capacity of PPDE in vitro was determined by scavenging experiments using DPPH, ABTS+·, ·OH, and ·O2-. The cell survival rate was determined by MTT assay. The MDA, SOD, CAT, GSH, and GSH-Px content were determined by colorimetry assays. The expression levels of antioxidant genes SOD, CAT, GSH, and GSH-Px were assessed by reverse transcription-quantitative PCR. HPLC was used to identify the PPDE components. The results suggested that PPDE had scavenging effects on DPPH, ABTS, hydroxyl, and superoxide anion radicals in a concentration-dependent manner; H2O2 treatment resulted in oxidative stress in LLC-PK1 cells, and the degree of injury of LLC-PK1 cells following PPDE treatment was improved, which was positively correlated with its concentration. Peucedanum praeruptorum Dunn extract treatment reduced the content of MDA and increased the content of CAT, SOD1, GSH, and GSH-Px. The mRNA expression levels of antioxidant genes detected by quantitative PCR were consistent with changes in CAT, SOD, GSS, and GSH-Px. Additionally, the trend in CAT, SOD1, GSH, and GSS protein expression levels was also consistent at the mRNA level. PPDE was found to consist of isochlorogenic acid C, myricetin, baicalin, luteolin, and kaempferol. Therefore, PPDE, which was formed of products derived from natural substances, functioned in the inhibition of oxidative damage. The present study aimed to obtain a better understanding of the traditional Chinese medicine Peucedanum praeruptorum Dunn and preliminarily elucidate its antioxidant mechanism at the cellular level. Further animal or human experiments are required to verify the antioxidant effects of PPDE for further development and utilization.

Cynapanoside A exerts protective effects against obesity-induced diabetic nephropathy through ameliorating TRIM31-mediated inflammation, lipid synthesis and fibrosis.

Obesity is a major predictive factor for the diabetic nephropathy (DN). However, the precise mechanism and therapeutic approach still require to be investigated. Cynapanosides A (CPS-A) is a glycoside derived from the Chinese drug Cynanchum paniculatum that has numerous pharmacological activities, but its regulatory function on obesity-induced kidney disease is still obscure. In the present study, we attempted to explore the renoprotective effects of CPS-A on the established DN in high fat diet (HFD)-fed mice, and the underlying mechanisms. We initially found that CPS-A significantly ameliorated the obesity and metabolic syndrome in mice with HFD feeding. Mice with HFD-induced DN exerted renal dysfunctions, indicated by the elevated functional parameters, including up-regulated blood urea nitrogen (BUN), urine albumin and creatinine, which were significantly attenuated by CPS-A in obese mice. Moreover, histological changes including glomerular enlargement, sclerosis index and collagen deposition in kidney of obese mice were detected, while being strongly ameliorated by CPS-A. Additionally, podocyte loss induced by HFD was also markedly mitigated in mice with CPS-A supplementation. HFD feeding also led to lipid deposition and inflammatory response in renal tissues of obese mice, whereas being considerably attenuated after CPS-A consumption. Intriguingly, we found that tripartite motif-containing protein 31 (TRIM31) signaling might be a crucial mechanism for CPS-A to perform its renoprotective functions in mice with DN. The anti-inflammatory, anti-fibrotic and anti-dyslipidemia capacities of CPS-A were confirmed in the mouse podocytes under varying metabolic stresses, which were however almost abolished upon TRIM31 ablation. These data elucidated that TRIM31 expression was largely required for CPS-A to perform its renoprotective effects. Collectively, our study is the first to reveal that CPS-A may be a promising therapeutic strategy for the treatment of obesity-induced DN or associated kidney disease.

ZEB1 promotes invasion and metastasis of endometrial cancer by interacting with HDGF and inducing its transcription.

Zinc finger E-box binding homeobox 1 (ZEB1), as a typical transcription inhibitory factor of E-cadherin, plays a major role in stimulating the invasion and metastasis of tumors via modulating the epithelial-mesenchymal transition (EMT) signal. However, its function and modulatory mechanisms in endometrial carcinoma (EC) remain unclear. In this study, silencing ZEB1 significantly reduced EC cell migration, invasion, and metastasis, as well as enhanced the sensitivity of EC cells to cisplatin (cDDP) in vitro and in vivo. Mechanism analysis indicated that ZEB1 interacts with hepatoma-derived growth factor (HDGF) and co-localizes in the nucleus. In addition, ZEB1 as a transcription factor binds to the promoter of HDGF to stimulate HDGF transcription. Furthermore, suppression of HDGF in ZEB1-overexpressed EC cells not only reduced the expression of ß-catenin, TCF4, and ZEB1, but also repressed ß-catenin translocation from the cytoplasm into the nucleus and further downregulated the combination with TCF4. Interestingly, the ß-catenin/TCF4 signaling feedback stimulates ZEB1 transcription and therefore constitutes a positive feedback loop. In clinical samples, ZEB1 positively correlates with HDGF expression, and co-expression of ZEB1 and HDGF promotes the pathogenesis of EC. In summary, our study demonstrated that the positive feedback loop of ZEB1/HDGF/ß-catenin/TCF4 plays an unfavorable role in the metastasis of endometrial carcinoma.

Gardenia jasminoides has therapeutic effects on L­NNA­induced hypertension in vivo.

Gardenia jasminoides is a plant that has been used in traditional Chinese medicine. It has four key active components (genipin gentiobioside, geniposide, crocin 1 and crocin 2). The aim of the present study was to determine the anti­hypertension effects of Gardenia jasminoidesin vivo. The chemical composition of Gardenia jasminoides was determined using liquid chromatography. The anti­hypertensive effects of Gardenia jasminoides were determined by a L­NG­nitroarginine (L­NNA)­induced hypertension animal model. Both Gardenia jasminoides plants of the Jiangjin County variety (CJGJ) and the Lichuan City variety (HLGJ) were used. HLGJ contained more geniposide than CJGJ. L­NNA was used to induce hypertension in mice, and the mice were subsequently treated with CJGJ and HLGJ. The Gardenia jasminoides­treated mice exhibited lower systolic (SBP), diastolic (DBP) and mean blood pressure (MBP) than the experimental control mice. Additionally, HLGL has a more potent effect on SBP, MBP and DBP than CJGJ. Following Gardenia jasminoides treatment, the nitric oxide contents in serum, heart, liver, kidney and stomach of mice were higher than the L­NNA­induced control mice, and the malondialdehyde contents were lower; the levels in HLGJ­treated mice were closer to those normal mice than the levels in CJGJ­treated mice were. Serum levels of endothelin­1 and vascular endothelial growth factor were reduced by HLGJ treatment in hypertensive mice, whereas the calcitonin gene­related peptide level was raised. Reverse transcription­polymerase chain reaction analysis of mouse heart and vessel tissue demonstrated that HLGJ­treated mice exhibited higher heme oxygenase­1, neuronal nitric oxide synthase (nNOS), endothelial NOS, Bax, caspase­3, caspase­8, caspase­9 mRNA expression levels and lower adrenomedullin, receptor activity modifying protein, interleukin­1ß, tumor necrosis factor­α, inducible NOS, Bcl­2, monocyte chemoattractant protein­1, nuclear factor­κB and matrix metalloproteinase­2 and ­9 mRNA expression compared with control hypertensive mice and CJGJ­treated mice. In conclusion, Gardenia jasminoides has anti­hypertensive effects, and these effects may be associated with the active component, geniposide.

Preventive effect of Gardenia jasminoides on HCl/ethanol induced gastric injury in mice.

The therapeutic effect on HCl/ethanol induced gastric injury of Gardenia jasminoides (JXGJ-1 and JXGJ-2) were determined by a animal model. JXGJ-2 group reduced area of its gastric injury as compared to the control group, JXGJ-2 also helped in decreasing the gastric secretion volume results raised in pH value. The NO contents in serum, heart, liver, kidney and stomach of JXGJ-2 group were more than JXGJ-1 and control groups. JXGJ-2 reduce cytokine levels as compared to JXGJ-1 and control group. The serum and gastric tissue SOD, GSH-Px, GSH levels in JXGJ-2 treated mice were higher than JXGJ-1 treated and control mice, but the MDA, PC levels showed the crosscurrents, these levels were close to normal mice. Gardenia jasminoides could increase the occludin, EGF, EGFR, VEGF, IκB-α, nNOS, eNOS, Cu/Zn-SOD, Mn-SOD, CAT, GSH-Px (GSH1) mRNA and protein expressions and decrease the p38MAPK (p38), NF-κB, Bcl-2, COX-2, iNOS expressions in gastric tissues unlike to the control mice, JXGJ-2 had much better effect than JXGJ-1. JXGJ-1contained the higher genipin gentiobioside and gardenoside, they might be the key components of gastric injury inhibition. Gardenia jasminoides had a remarkable effect on gastric injury, and they were derived from two important components of genipin gentiobioside and gardenoside.

Preventive effect of polysaccharides from the large yellow croaker swim bladder on HCl/ethanol induced gastric injury in mice.

In the present study the preventive effect of polysaccharides from the large yellow croaker swim bladder (PLYCSB) on HCl/ethanol-induced gastric injury in ICR mice was investigated. A high dose of PLYCSB (50 mg/kg) was found to reduce the levels of the serum proinflammatory cytokines interleukin (IL)-1ß, IL-6, IL-8, as well as increase the levels of IL-4 compared with those in mice treated with a low dose of PLYCSB (25 mg/kg) and control mice. The somatostatin and vasoactive intestinal peptide serum levels in PLYCSB-treated mice were higher compared with those in control mice, whilst motilin and substance P serum levels were lower compared with those in control mice. The extent of the gastric injury in the mice treated with PLYCSB was lower compared with that in the control mice; however, the results obtained for mice treated with a high dose of PLYCSB were similar to those for omeprazole-treated mice. In addition, the superoxide dismutase and glutathione peroxidase activities of PLYCSB-treated mice were higher compared with those of the control mice, and similar to those observed in normal and omeprazole-treated mice. Furthermore, PLYCSB-treated mice showed levels of nitric oxide and malondialdehyde that were similar to those in the normal group. Using PCR and western blot analysis, it was demonstrated that PLYCSB significantly inhibited inflammation in the tissues of the HCl/ethanol induced gastric injury mice by downregulating the expression of inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-α and IL-1ß. These results suggest that PLYCSB has an inhibitory effect against gastric injury that is comparable to that of the gastric injury drug omeprazole. Therefore, PLYCSB has the potential to be used as a natural therapeutic drug.

Integrating Computational Modeling and Experimental Assay to Discover New Potent ACE-Inhibitory Peptides.

Human angiotensin-I-converting enzyme (ACE) is an important target of antihypertensive therapy, which possesses a bulky, hydrophobic pocket that is physicochemically compatible with a wide variety of peptide substrates and small-molecule ligands. Rational design of potent ACE inhibitors has long been an attractive topic in the chemical, biological and medical communities. In the present study, an integrative protocol is described to optimize and modify peptides bound with ACE based on their complex three-dimensional structures. The protocol combines a number of sophisticated computational methods including molecular dynamics (MD) simulation, quantitative structure-activity relationship (QSAR), molecular mechanics/Poisson-Boltzmann surface area (MM-PB/SA) and quantum mechanics/molecular mechanics (QM/MM) to discover new potent ACE-inhibitory peptides. With this strategy hundreds of potential peptides are generated virtually, from which several promising candidates are synthesized and assayed in vitro using a standard Fmoc -protected amino acid solid phase synthesis and spectrophotometric method, respectively. Six peptides are found to have potency against ACE, four of which (LVY, VLKP, MLPVY and LKIPLY) show satisfactory inhibitory capability (pIC50 =5.84, 5.27, 5.40 and 5.57, respectively). Subsequently, the complex structures of tripeptide LVY and hexapeptide LKIPLY with ACE are computationally modeled and their binding free energies are estimated as - 34.2 and - 57.8 kcal/mol, respectively, by using a rigorous QM/MM scheme. Intensive steric collisions are observed around the C-terminus of peptide ligands in the bound state. Based upon the findings the C-terminal residues of the two peptides are modified to small, hydrophobic amino acids Val and Ala, resulting in mutated peptides LVA and LKIPVA, respectively. Consequently, a significant improvement in ACE-inhibitory activity is observed for the LKIPVA mutant (pIC50 increases from 5.57 to 6.07), whereas only a modest activity change was associated with the LVA mutant (pIC50 decreases from 5.84 to 5.80).

Antitumor activities of Juemingzi (Cassia tora L.) on Balb/c sarcoma 180-injected mice.

The antitumor activity of Juemingzi (Cassia tora L.) was investigated in mice that were fed various concentrations of the compound. Although mice fed a low concentration (50 mg/kg b.w.) of Juemingzi exhibited a high tumor weight, the higher feeding concentrations (100 and 200 mg/kg b.w.) were associated with lower weight tumors. The growth rate of mouse splenocytes that were treated with 200 mg/kg b.w. Juemingzi was determined using a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide assay. This rate of proliferation was higher than that achieved with 100 and 50 mg/kg b.w. Juemingzi treatment by fetal bovine serum, lipopolysaccharide or concanavalin A. Compared with the lower concentrations of Juemingzi treatment, 200 mg/kg b.w. Juemingzi significantly (P<0.05) reduced aspartate aminotransferase, alanine transaminase and blood urea nitrogen levels. A high concentration of Juemingzi (200 mg/kg b.w.) significantly (P<0.05) increased the levels of tumor necrosis factor-α and interleukin-1ß cytokines compared with those of the mice that were treated with 100 and 50 mg/kg b.w. Juemingzi.

[Study on species and distribution of flora of national rare and endangered medicinal plant in the Three Gorges area].

According to the China Plant Red Data Book and National Key Protected Wild Plants, the distribution of the rare and endangered plants and national conservative plants in the Three Gorges area were investigated and statistically analyzed. Its floristic composition and characteristics of geographical distribution were explored. As a result, a total of 97 species of medicinal flora belonging to rare and endangered national protection plants were found in the Three Gorges area. They come from 81 genera of 46 families. Their vertical distribution is obvious and horizontal distribution has discontinuous overlap. There are many ancient relict medicinal plants in the Three Gorges area. These medicinal plants have obvious temperate characteristics, and are easily found at warm and moist ravines and hillsides; The proportion of tree is much higher than that of herb, vine, shrub and fern. Most of them belong to specific and monotypic genera.

Preparation and evaluation of a new releasable PEGylated tumor necrosis factor-α (TNF-α) conjugate for therapeutic application.

To design a releasable PEGylated TNF-α (rPEG-TNF-α), a cathepsin B-sensitive dipeptide (Val-Cit moiety) was inserted into conventional PEG-modified TNF-α (PEG-TNF-α), facilitating its clinical use for anti-tumor therapy. Comparative pharmacokinetic and pharmacodynamic studies showed that the half-lives of both PEGylated forms of TNF-α were ∼60-fold greater than that of unmodified TNF-α. In addition, the in vitro bioactivity of rPEG-TNF-α was greater than that of PEG-TNF-α with the same degree of PEG modification. Release of TNF-α from rPEG-TNF-α in vitro was dependent on the presence of cathepsin B and was inhibited by a cathepsin B inhibitor. Despite the potent cytotoxicity of unmodified TNF-α against normal cells, its PEGylated forms at higher TNF-α concentrations showed low cytotoxic activity against these cells. In contrast, both forms of PEGylated TNF-α showed potent cytotoxic activity against the B16 and L929 cell lines, with rPEG-TNF-α being 5- and 9-fold more potent, respectively, than PEG-TNF-α. Moreover, rPEG-TNF-α was a more potent in vivo antitumor agent than PEG-TNF-α.