Diabetic nephropathy (DN), a common complication associated with both type I and type II diabetes mellitus (DM), is a major cause of chronic nephropathy and a common cause of end-stage renal diseases (ESRD) throughout the world. This study is aimed to determine whether dencichine (De) can ameliorate renal damage in high-glucose-and-fat diet combined STZ (streptozocin) induced DN in type II DM rats and to investigate the potential underlying mechanisms. Markers of metabolism, diabetes, and renal function, and levels of extracellular matrix (ECM) collagen I (Col I), collagen IV (Col IV), fibronectin (FN) and laminin (LN), and of proteins in the TGF-β/Smad pathway were analysed through RT-PCR, western blot, immunofluorescence and immunohistochemistry. The results show that De significantly alleviates metabolism disorder, improved renal function, relieved pathological alterations in the glomerulus of DN rats, decreased ECM deposition and increased the ratio of matrix metalloproteinase (MMP)-9 to tissue inhibitor of metalloproteinase (TIMP)-1 both in vivo and in vitro. Moreover, De negatively regulated TGF-β/Smad signalling pathway and increased the expression of Smad7, an endogenic inhibitory Smad located downstream of the signalling pathway. In conclusion, we provide experimental evidence indicating that the renoprotective effect of De could significantly prevent the progression of DN possibly attribute to down-regulation of the TGF-β/Smad pathway and rebalance the deposition and degradation of ECM proteins.Download high-res image (227KB)Download full-size image

The purpose of our research is to find a new lipid emulsion to deliver a low water-soluble compound, cinnamaldehyde (CA). Its characteristics, pharmacokinetics, antitumor efficacy, and toxicity were evaluated. The mean particle size, zeta potential, and encapsulation efficiency of the submicromemter emulsion of CA (SME-CA) were 130 ± 5.92 nm, −25.7 ± 6.00 mV, and 99.5 ± 0.25%, respectively. The area under the curve from 0 h to termination time (AUC0–t) of SME-CA showed a significantly higher value than that of CA (589 ± 59.2 vs 375 ± 83.5 ng h/L, P < 0.01). Tissue distribution study showed various changes; among them, a 27% higher concentration was found in brain tissue when using SME-CA at 15 min after administration. For the efficacy evaluation, SME-CA exhibited 8- and 11-fold antitumor activity in the depression of HeLa and A549 cell lines with the IC50 decreasing to 0.003 and 0.001 mmol/L, respectively. The LD50 values of CA and SME-CA in mice were 74.8 and 125 mg/kg, suggesting increased safety from the new formulation. The new formulation exhibited lower toxicity, higher antitumor activity, and a more satisfactory pharmacokinetic property, which displayed great potential for future pharmacological application.

In this study, a novel, convenient, accurate, and valid method was developed by using high-performance liquid chromatography-photodiode array detection to obtain a chromatographic fingerprint of Antike capsule (AC). Using computer aided similarity evaluation software, 28 characteristic peaks in chromatograms of 10 batches of analyzed samples were screened out and traced to the source of original materials, toad skin and angelica, in which 16 of the peaks were identified as gamabufotalin, arenobufagin, telocinobufagin, desacetylcinobufotalin, bufotalin, cinobufotalin, bufalin, cinobufagin, resibufogenin, ferulic acid, n-butylidenephthalide, senkyunolide A, senkyunolide I, senkyunolide H, ligustilide, and coniferylferulate. At the same time, the fingerprint similarity was calculated and the contents of known ingredients were also determined simultaneously. This method demonstrated good precision, reproducibility, and stability (relative standard deviation [RSD] of relative retention time [RRT] < 2.0% and RSD of relative peak area [RPA] < 5.0%). Good linear behaviors over the investigated concentration ranges were observed for all the analytes (r2 > 0.9994), and the recoveries and RSD varied from 96.35% to 102.43% and 0.48% to 1.98%, respectively. The proposed method enabled fingerprint analysis and simultaneous identification and determination of 16 constituents in a single run. In addition, it provides a significant reference for the quality control of AC.

•A new method is described to simultaneously quantify cinnamaldehyde and its metabolites.•The pharmacokinetic parameters of cinnamaldehyde and its metabolites are determined.•A new metabolite methyl cinnamate was determined.•Pharmacokinetic study showed that a small portion of cinnamaldehyde sustained a low concentration for a long time.A selective and sensitive method utilizing gas chromatography–mass spectrometry was developed for simultaneous determination of cinnamaldehyde, cinnamyl alcohol, and methyl cinnamate in rat plasma. Cinnamaldehyde and cinnamyl alcohol can inter-convert to one another in rats, thus simultaneous quantifying both analytes provided a reliable and accurate method of assessment. Three qualifying ions (131 m/z, 105 m/z and 92 m/z) were chosen for simultaneous quantification of cinnamaldehyde and its metabolites. In this study, the calibration curves demonstrated a good linearity and reproducibility over the range of 20–2000 ng/ml (r2 ≥ 0.999) for all analytes. Furthermore, the sensitivity of gas chromatography–mass spectrometry revealed sufficient lower limit of quantitation and detection of 20 ng/ml and 5 ng/ml, respectively, in the pharmacokinetic analysis. The intra- and inter-day precision variations were less than 10.4% and 12.2%, respectively, whilst accuracy values ranged from −8.6% to 14.8%. All analytes were stable in plasma and in processed samples at room temperature for 24 h with no significant degradation after three freeze/thaw cycles. A small amount of the administered cinnamaldehyde had long half-life of 6.7 ± 1.5 h. In this study, gas chromatography–mass spectrometry was demonstrated to be a powerful tool for the pharmacokinetic studies of rats after intravenous and oral administration of cinnamaldehyde.

•A pectin-coated liposomal formulations was prepared by forming an ion-complex.•The pectin-coated liposome has an excellent stability and mucoadhesiveness properties.•The pectin-coated liposome would be a promising drug carrier system for colon cancer.The aim of this study was to investigate the potential effect of pectin for liposomal drug delivery systems. An orthogonal L9 (33) test was designed to optimize the preparation condition of cationic bufalin liposomes coated with commercially available citrus pectin (CPL). The change in particle size, zeta potential, entrapment efficiency, stability, mucoadhesion and anticancer effect were evaluated. The results showed that CPL had an excellent stability and mucoadhesive properties, and the drug release in vitro was modest prolonged and sustained. Furthermore, the inhibition effect of liposomes on SW480 colon cancer cells was dramatic enhanced due to a block of cell cycle at G0/G1 phase, and CPL had a higher inhibition rate than bufalin liposomes (BFL) because of the anticancer effect of citrus pectin. It is concluded that CPL is a potentially promising drug carrier system treatment for colon cancer.

Lung cancer as a malignance has been killing numerous patients around the world annually, and small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) are the two major types, the later accounting for nearly 80 % of lung cancer. There are multiple causes for lung cancer, and more researches have been carried out to prevent, anticipate, and diagnose the cancer. MicroRNAs (miRNAs) are small non-coding RNA molecules capable of regulating expression of over 50 % of protein-coding genes. The RNA molecules are stable in tissues and blood, so it can tend to be a biomarker in anti-lung cancer. Here, this is a review on the roles of miRNAs for possible ways to prevent lung cancer in clinical trials.

•Tetrahydroxystilbene glucoside (TSG) improved the memory ability of mice.•TSG reduced the serum levels of ROS, NO and IGF-1 and increased the levels of SOD, Ca2+ and Klotho protein.•TSG up-regulated the expression of Klotho protein in cerebrum, heart, kidney, testis and epididymis tissues.Tetrahydroxystilbene glucoside (TSG) is a strong antioxidant and free radical scavenger derived from Polygonum multiflorum Thunb. The present study aims to evaluate the protective effect of TSG against d-galactose induced aging process in mice and its possible mechanisms of action. Our study revealed that administration of TSG improved the memory ability and regulated the body weight of mice. TSG also reduced the levels of ROS, NO and IGF-1 and increased the levels of SOD, Ca2+ and Klotho protein in the serum. Furthermore, TSG up-regulated the expression of Klotho protein in cerebrum, heart, kidney, testis and epididymis tissues of d-galactose induced aging mice. These results suggested that TSG had a promising anti-aging effect by regulating Klotho gene.Tetrahydroxystilbene glucoside (TSG) exhibited significant protective effect against d-galactose induced aging process in mice by regulating Klotho gene.

A simple RP-LC-UV method was established for the determination of tryptanthrin in plasma and different tissues of rats. The separation was achieved by HPLC on a C18 column with a mobile phases composed of acetonitrile–water (47:53, v/v), UV detection was used at 251 nm. Good linearity was found between 0.0183–1.1712 μg mL−1 (r2 = 0.999) for plasma and 0.0937–1.7568 μg mL−1 for the tissue samples, respectively (r2 ≥ 0.9932). The intra- and inter-day precisions expressed as the relative standard deviation for the method were 0.92–6.01 and 1.06–9.11 %, respectively. The relative recoveries of tryptanthrin ranged from 95.26 to 97.89 % for plasma and 82.55 to 114.99 % for tissue homogenates (except heart). The developed method was successfully applied to the pharmacokinetics and tissue distribution research after orally administration of a 56-mg kg−1 dose of tryptanthrin to healthy SD rats. The main pharmacokinetics distribution results showed that liver, lung, small intestine, and large intestine were the major distribution tissues of tryptanthrin in rats, and that tryptanthrin had difficulty in crossing the blood–brain barrier.

Imperatorin (IMP) is a biologically active ingredient isolated from a traditional Chinese medicine (TCM), Angelica dahurica. To obtain the brain distribution data of IMP in rats, the concentrations of IMP in cortex, cerebellum, diencephalon, brain stem, striatum and hippocampus were measured by a simple and sensitive HPLC–UV method. The analytes were prepared by a liquid–liquid extraction method and the separation of IMP was performed on a Hypersil BDS C18 column (250 mm × 4.6 mm i.d., 5 μm) using acetonitrile–water (60:40, v/v) as mobile phase which was delivered at 1.0 mL min−1. Ultraviolet detection was performed at 300 nm. Using a weighted (1/c2) least square method, linear calibration curves for the six regions were obtained (r ≥ 0.9990) with a lower limit of quantification (LLOQ) of 0.075 μg g−1 for cortex, cerebellum, diencephalon and brain stem or 0.15 μg g−1 for striatum and hippocampus, and the recovery was greater than 90% for each tissue sample. The within- and between-day precisions (expressed as the relative standard deviation, RSD) were less than 10%. The validated method has been successfully applied to the brain distribution study in rats. The results showed that IMP could pass through the blood–brain barrier easily. And the higher concentration in striatum and hippocampus compared with the others might indicate that they were the target regions of IMP in rat brain.

•TMP inhibits tumor cell proliferation in HCC.•TMP induces apoptosis and autophagy in HCC.•ROS is involved in TMP-induced anticancer effects in HCC cells.Hepatocellular carcinoma (HCC) is one of the most common types of liver cancers with high recurrence rate and mortality rate. Recent studies have indicated that tetramethylpyrazine (TMP), a purified chemical extracted from Ligusticum wallichii Franchat (ChuanXiong), possessed antitumor effects on HCC, but detailed mechanism remains unclear. Our study aims at investigating the antitumor effect of TMP on HCC and its underlying mechanism. We found that TMP inhibited cell proliferation of HepG2 cells in a dose-dependent way, and xenograft tumor models also indicated that high concentrations of TMP administration inhibited tumor growth. Next, flow cytometric analysis and transmission electron microscope images showed that TMP enhanced cell apoptosis in HepG2 cells, and western blot results showed that TMP promoted cleavage of caspase-3 and PARP in vitro and in vivo. We also found that TMP caused autophagy in HCC in vitro and in vivo. In order to examine the role of autophagy in TMP-induced apoptosis, 3-methyladenine (3-MA) was used to block the action of autophagy. Our data showed TMP-induced autophagy might be a pro-apoptosis process in HCC. Furthermore, the results of anti-oxidative enzymes and oxidation-sensitive fluorescent probe 2, 7-dichlorofluorescein diacetate (DCFH-DA) indicated that TMP induced ROS generation and inhibition of ROS diminished the anticancer function of TMP. In conclusion, our studies provide new insights into the mechanisms underlying the antitumor effect of TMP and suggest that TMP can be a novel therapeutic regimen for HCC.

•CA attenuate ROS production and autophagy which result in cardiac protection and cell survival.•NOX4 but not NOX2 was significantly enhanced after LPS stimulation and suppressed by CA.•CA exhibit anti-inflammation activity by directly suppress TLR4 and NOX4.•Homology modeling and molecular docking was used to imitate the conjunction of CA with both TLR4 and NOX4.Cinnamaldehyde (CA), a major bioactive compound extracted from the essential oil of Cortex Cinnamomi, exhibits anti-inflammatory activity on endotoxemia. Accumulating evidence indicates reactive oxygen species (ROS) and autophagy play a vital role in the cardiac dysfunction during endotoxemia. The aim of this study was to unveil the mechanism of CA on ROS production and autophagy during endotoxemia. Male Sprague-Dawley rats were stimulated by LPS (20 mg/kg i.v.) with or without treatment of CA. Cardiac function and histopathological staining were preformed 4 h after LPS stimulation. The levels of TNF-α, IL-1β and IL-6 were detected by ELISA. The expression of p-JNK, p-ERK, p-p38, TLR4, NOX4, NOX2, ATG5 and LC3 proteins were determined by Western blot. The results showed that CA inhibited cardiac dysfunction, inflammatory infiltration and the levels of TNF-α, IL-1β and IL-6 in LPS stimulated rats by blocking the TLR4, NOX4, MAPK and autophagy signalings. In order to obtain further confirmation of the mechanism of CA on endotoxemia in vitro, a limited time-course study was firstly performed by Western blot. TLR4, NOX4 and LC3 were significantly increased after 4 h LPS stimulation. CA reversed the intracellular ROS production and MAPK signaling activation induced by LPS. Electron microscopy, mRFP-GFP-LC3 transfection and western blot results revealed autophagic flux were attenuated after CA treatment. The siRNA and molecular docking results suggest that CA can suppress both TLR4 and NOX4 during endotoxemia. Our data revealed that CA ameliorated LPS-induced cardiac dysfunction by inhibiting ROS production and autophagy through TLR4-NOX4 pathway.Download high-res image (143KB)Download full-size image

In Europe, swainsonine has been studied widely for prevention of metastasis and cancer therapy. In order to investigate the effects and mechanisms of swainsonine on the human gastric carcinoma SGC-7901 cell, we carried out in vivo and in vitro experiments.After treatment with swainsonine, an effective dose and IC50 value of swainsonine for SGC-7901 cells were examined by MTT assay. Cell-cycle distribution and apoptotic rates were analyzed using FCM, and [Ca2+]i was measured using LSCM. The expression of p53, c-myc and Bcl-2 were determined using an immunocytochemical method. Simultaneously, 50 mice were divided randomly into five groups. Three groups were administrated swainsonine at dose of 3, 6 and 12 mg/kg body wt., two control groups were administrated N.S. 20 ml/kg body wt. and 5-Fu 20 mg/kg body wt., respectively, by intraperitoneal injection. The inhibition rate was calculated and pathological sections were observed.The growth of SGC-7901 cell is inhibited by swainsonine in vitro, with an IC50 value at 24 h of 0.84 μg/ml, and complete inhibition concentration is 6.2 μg/ml. After treatment with swainsonine at the concentrations of 0.5, 1.5 and 4.5 μg/ml for 24 h, the expression of apoptosis inhibiting gene p53 and bcl-2 decreases, and the apoptotic trigger gene c-myc increases markedly (p<0.05), as well as [Ca2+]i overloading, SGC-7901 cell is induced to apoptosis in the end. It is also found that the percentages of S phase are 38.8%, 39.7% and 29.6%, respectively (20.0% in control group and 23.2% in 5-Fu group). The rates of inhibition were 13.2%, 28.9%, 27.3%, respectively, when the nude mice were administered swainsonine (p<0.05 or 0.01). The structure of the tumor showed hemorrhage, necrosis and inflammatory cell infiltration. We therefore conclude that swainsonine could inhibit cell proliferation in vitro and the growth of human gastric carcinoma in vivo. The mechanisms of swainsonine-induced apoptosis may relate to [Ca2+]i overloading and the expression of apoptosis-related genes.

AimTo examine the efficacy of YuanHu painkillers (YHP) as a treatment for primary dysmenorrhea and to reveal YHP's principle formula.MethodsA Wistar rat uterine contraction model was utilized in this study. Rats were given 0.698 g/kg YHP, 0.07 g/kg tetrahydropalmatine (THP; YHP's main component), 0.02 g/kg imperatorin (IMP), or THP + IMP (0.07 + 0.02 g/kg) as polypharmacy (PG) by gavage. H&E staining and histopathological examination of the uteri tissue samples were performed. We then detected superoxide dismutase (SOD) and malondialdehyde (MDA), nitric oxide (NO), as well as inducible nitric oxide synthase (iNOS), i-κB, nuclear factor-κB (NF-κB), and cyclooxygenase-2 (COX-2) indices.ResultsPG significantly inhibited the uterine contraction of the primary dysmenorrhea rat model (p < 0.05), and was significantly different than single-agent therapy (p < 0.05). Histopathological examination showed inflammation in the uteri of the control group which YHP and its main constitutes alleviated. THP significantly inhibited the contraction of isolated uteri caused by Ach, PGF2α and oxytocin in a concentration-dependent fashion. THP and IMP both significantly affected the levels of NO, activation of NF-κB, up-regulated the expression of i-κB and down-regulated the expression of both iNOS and COX-2. IMP obviously decreased the level of MDA and increased the activation of SOD (p < 0.05). PG obviously improved all the parameters mentioned above (p < 0.05).ConclusionsYHP exerted protective effects on primary dysmenorrhea in rats and remarkably alleviated the severity of experimental primary dysmenorrhea. The combined strategy proved to be more effective than either THP or IMP alone and may have synergistic effects in combination in primary dysmenorrhea. Mechanisms that might account for the beneficial effects include abating oxidative stress, inhibiting over-inflammatory reaction, and alleviating the contraction of isolated rat uteri by inhibiting the influx of extracellular Ca2+. Broad potential for future clinical practice is foreseeable.

Polydatin preconditioning (PPC) has been reported to be protective against brain and intestine ischemia/reperfusion injury (I/R injury), but whether polydatin exerts cardioprotective effect against myocardial ischemia/reperfusion and the underlying mechanisms remain unclear. Previous studies have demonstrated that oxidative stress plays an important role in the process of I/R. Elevation of oxidative agents and decline in anti-oxidant substance would promote I/R. Meanwhile, the activation of PKC signaling seems to mediate the cardioprotective effects of many drugs by alleviating Ca2+ influx. In the present study, we reported for the first time that intravenous administration of polydatin before I/R significantly limited the infarct size, creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) leakage from the damaged myocardium after I/R. The activity of SOD and the content of MDA remarkably changed in the presence of polydatin as well. However, the cardiac function-preserving and myocardial enzymes leakage-limiting effects of polydatin vanished in the presence of PKC inhibitors and mito KATP channel blockers. But there was not a significant change in the activity of SOD and MDA content. We therefore conclude that PPC exerts cardioprotective effect by the activation of PKC-KATP-dependent signaling and the direct anti-oxidative stress mechanisms.

AimTo study the effects of 2, 3, 5, 4′-tetrahydroxystilbene-2-O-β-d-glucoside (THSG) on proliferation of rat cardiac stem cells (CSCs) in vitro.Materials and methodsC-kit+ cells were isolated from neonatal (1 day old) Sprague–Dawley rats by using flow cytometry. Optimal THSG treatment times and doses for growth of CSCs were analyzed. CSCs were treated with various THSG doses (0, 1, 10, and 100 μM) for 12 h.ResultsSorted c-kit+ cells exhibited self-renewing and clonogenic capabilities. Cell Counting Kit (CCK-8) and Proliferating Cell Nuclear Antigen (PCNA) ELISA test positive cells were significantly increased in THSG-treated groups compared with untreated controls. The percentage of S-phase cells also increased after THSG treatment. Moreover, we show that some c-kit+ cells spontaneously express vascular endothelial growth factor (VEGF), T-box transcription factor (Tbx5), hyperpolarization-activated cyclic nucleotide-gated 2 (HCN2), hyperpolarization-activated cyclic nucleotide gated 4 (HCN4), alpha myosin heavy chain (αMHC), and beta myosin heavy chain (βMHC) mRNA, and stem cell antigen 1 (Sca-1), cardiac troponin-I, GATA-4, Nkx2.5, and connexin 43 protein were also assessed in CSCs. However, their expression was significantly increased with THSG treatment when compared to untreated controls.ConclusionTHSG can increase proliferation of rat CSCs in vitro and thus, shows promise as a potential treatment strategy for stimulating endogenous stem cells to help repair the injured heart after myocardial infarction in patients.

Headings aimsCardiac stem cells (CSCs)-transplanted therapy provides a promising therapy for the ischemic heart disease (IHD), especially in the epidemic of myocardial infarction (MI). The compound 2,3,5,4′-tetrahydroxystilbene-2-O-β-d-glucoside (THSG) can induce CSC proliferation in vitro based on our previous study, so we aimed to study the induce effect of THSG on CSCs-transplanted MI rat in vivo.Materials and methodsUsing a murine model of MI, this study was designed to evaluate the impact of THSG (30, 60, 120 mg/kg) on CSCs-based therapy for MI and the underlying mechanism in this process.Key findingThe results showed that THSG on CSCs-transplanted therapy groups (THSG + CSCs groups) can significantly reduce S-T segment elevation, and increase heart rate compared with MI group. The left ventricular ejection fraction (LVEF) and the left ventricular fractional shortening (LVFS) were significantly reduced in THSG + CSCs groups compared to the MI group. The levels of enzyme expression (CK-MB, LDH), the heart weight index (HWI) and myocardial infarct size (IS) were all reduced in THSG + CSCs groups. Moreover, other changes noted during these 28 days post-MI, included pathologic changes, as well as increased stem cell antigen-1 (Sca-1) expression, or expression of Nkx2.5, GATA-4, and Connexin 43 in myocardial tissue, and reduced the Caspase-3 expression.SignificanceOur findings indicated that THSG facilitated CSCs-transplanted therapy in MI. These observations may be associated with the inducted of THSG on the proliferation of CSCs in vivo and also, with the subsequent differentiation of additional intrinsic neonatal cardiomyocytes to replace damaged heart tissue.

In this study, a novel, convenient, accurate, and valid method was developed by using high-performance liquid chromatography-photodiode array detection to obtain a chromatographic fingerprint of Antike capsule (AC). Using computer aided similarity evaluation software, 28 characteristic peaks in chromatograms of 10 batches of analyzed samples were screened out and traced to the source of original materials, toad skin and angelica, in which 16 of the peaks were identified as gamabufotalin, arenobufagin, telocinobufagin, desacetylcinobufotalin, bufotalin, cinobufotalin, bufalin, cinobufagin, resibufogenin, ferulic acid, n-butylidenephthalide, senkyunolide A, senkyunolide I, senkyunolide H, ligustilide, and coniferylferulate. At the same time, the fingerprint similarity was calculated and the contents of known ingredients were also determined simultaneously. This method demonstrated good precision, reproducibility, and stability (relative standard deviation [RSD] of relative retention time [RRT] < 2.0% and RSD of relative peak area [RPA] < 5.0%). Good linear behaviors over the investigated concentration ranges were observed for all the analytes (r2 > 0.9994), and the recoveries and RSD varied from 96.35% to 102.43% and 0.48% to 1.98%, respectively. The proposed method enabled fingerprint analysis and simultaneous identification and determination of 16 constituents in a single run. In addition, it provides a significant reference for the quality control of AC.