ObjectiveThis study aims to investigate the therapeutic effects of Wenweishu granule （WWSG） on functional dyspepsia （FD） with spleen-stomach deficiency cold syndrome in rats by integrating network pharmacology， molecular docking， and animal experiments. MethodsActive components and corresponding targets of WWSG were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and the Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine （BATMAN-TCM）. Disease-related targets for FD with spleen-stomach deficiency cold syndrome were screened using GeneCards and the Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP）. Core therapeutic targets were identified via Cytoscape and validated by molecular docking. A rat model of FD with spleen-stomach deficiency cold syndrome was established using vinegar gavage combined with tail-clamping. The rats were randomly divided into a model group， low-， medium-， and high-dose WWSG groups （2.0， 4.0， 8.0 g·kg^-1）， a domperidone group （3.0 mg·kg^-1）， a Fuzi Lizhong pillwan （0.8 g·kg^-1）， and a normal control group （n=10 per group）. Drugs were administered once daily by gavage for 14 consecutive days. After treatment， body weight， symptom scores， and gastrointestinal motility indices were recorded. Gastric and duodenal pathologies changes were observed via hematoxylin-eosin （HE） staining. Brain-gut peptides were measured in serum and tissue using enzyme-linked immunosorbent assay （ELISA）. Immunohistochemistry and Western blot were performed to assess stem cell factor （SCF） and receptor tyrosine kinase （c-Kit） protein expression in gastric tissues. ResultsA total of 305 drug targets， 1 140 disease targets， and 116 overlapping targets were identified. Cytoscape analysis revealed 104 core targets. Enrichment analysis indicated that the SCF/c-Kit signaling pathway was the key mechanism. Molecular docking confirmed a strong binding affinity between active components of WWSG and SCF/c-Kit proteins （binding energy<-5.1 kcal·mol^-1）. Compared with the normal group， model rats exhibited slower weight gain （P<0.05）， reduced gastric emptying and intestinal propulsion （P<0.01）， mild gastric mucosal shedding， duodenal inflammatory cell infiltration， decreased levels of gastrin （GAS）， 5-hydroxytryptamine （5-HT）， and vasoactive intestinal peptide （VIP） （P<0.05， P<0.01）， and elevated somatostatin （SS） expression （P<0.05， P<0.01）. WWSG treatment ameliorated weight gain， symptom scores， and low-grade inflammation in gastric/duodenal tissues. High-dose WWSG significantly improved gastric emptying and intestinal propulsion， upregulated GAS， 5-HT， and VIP， and downregulated SS expression in serum and tissues （P<0.05， P<0.01）. Immunohistochemistry and Western blot demonstrated that SCF and c-Kit protein expression was decreased in the model group （P<0.05， P<0.01）， which was reversed by WWSG intervention （P<0.05）. ConclusionWWSG exerts therapeutic effects on FD with spleen-stomach deficiency cold syndrome in rats， potentially by regulating the SCF/c-Kit signaling pathway to enhance gastrointestinal motility.

Cyclodextrin metal-organic framework (CD-MOF) as a highly porous supramolecular carrier could be one of the solutions to the insolubility of isosteviol (STV). The solubility of STV was lower than 20.00 ng/mL at pH 1.0 and pH 4.5, whilst its solubility increased to 20,074.30 ng/mL at pH 6.8 and 129.58 ng/mL in water with a significant pH-dependence. The

Phytantriol (PT), ethanol (ET) and water were used to prepare in situ cubic liquid crystal (ISV2). The pseudo-ternary phase diagram of PT-ET-water was constructed and isotropic solution formulations were chosen for further optimization. The physicochemical properties of isotropic solution formulations were evaluated to optimize the composition of ISV2. In situ hexagonal liquid crystals (ISH2) were prepared based on the composition of ISV2 with the addition of vitamin E acetate (VitEA) and the amount of VitEA was optimized by in vitro release behavior. The phase structures of liquid crystalline gels formed by ISV2 and ISH2 in excess water were confirmed by crossed polarized light microscopy and small angle X-ray scattering, respectively. Rheological properties of ISV2 and ISH2 were studied by a DHR-2 rheometer. In vitro drug release studies were conducted by using a dialysis membrane diffusion method. Pharmacokinetics was investigated by determination of sinomenine hydrochloride (SMH) concentration in synovial membrane after intra-articular injection of SMH-loaded ISH2 in adjuvant-induced arthritis rats. The optimal ISV2 (PT/ET/water, 64 : 16 : 20, w/w/w) loaded with 6 mg x g(-1) of SMH showed a suitable pH, injectable and formed a cubic liquid crystalline gel in situ with minimum water absorption in the shortest time. The optimal ISV2 was able to sustain the drug release for 144 h. The optimal ISH2 system was prepared by addition of 5% VitEA into PT in the optimal ISV2 system. This ISH2 (PT/VitEA/ET/water, 60.8 : 3.2 : 16 : 20, w/w/w/w) was an injectable isotropic solution with suitable pH. The new ISH2 was able to sustain the drug release for more than 240 h. Local pharmacokinetics study indicated that the retention time and AUC(0-∞) of ISH2 group were increased significantly compared with that of SMH solution group and the AUC(0-∞) of ISH2 group was 6.01 times higher than that of SMH solution group. The developed ISH2 was suitable for intra-articular injection that may apply to patients in the treatment of rheumatoid arthritis.

The secretion of melatonin (MT) is obviously different in the younger and the senior sectors of the population, and the maximum plasma concentration of seniors is only half of that in the younger population group. If exogenous MT can be supplied to senior citizens based on the secretion rate and amount of endogenous MT in the younger population by a bio-mimetic drug delivery system (DDS), an improved therapeutic effect and reduced side effects can be expected. Based upon this hypothesis, the pharmacokinetic parameters of MT, namely, the absorption rate constant (k a), the elimination rate constant (k e), and the ratio of absorption rate (F) to the apparent volume of distribution (V) were obtained by a residual method depending on the plasma concentration curve of immediate release preparations in the healthy younger population. The dose-division method was applied to calculate the cumulative release profiles of MT achieved by oral administration of a controlled release drug delivery system (DDS) to generate plasma MT profiles similar to the physiological level-time profiles. The in vivo release of MT deduced from the healthy younger population physiological MT profiles as the pharmacokinetic output of the bio-mimetic DDS showed a two-phase profile with two different zero order release rates, namely, 4.919 μg/h during 0-4 h (r=0.9992), and 11.097 μg/h during 4-12 h (r=0.9886), respectively. Since the osmotic pump type of DDS generally exhibits a good correlation between in vivo and in vitro release behaviors, an osmotic pump controlled delivery system was designed in combination with dry coating technology targeting on the cumulative release characteristics to mimic the physiological MT profiles in the healthy younger population. The high similarity between the experimental drug release profiles and the theoretical profiles (similarity factor f 2>50) and the high correlation between the predicted plasma concentration profiles and the theoretical plasma concentration profiles (r=0.9366, 0.9163, 0.9264) indicated that a prototype bio-mimetic drug delivery system of MT was established. The similarity factors between the experimental drug release profiles and the theoretical release profile were all larger than 50 both in periods of 0-4 h and 4-12 h, namely, 68.8 and 57.3 for the first batch (Batch No. 20131031), 76.7 and 50.2 for the second batch (Batch No. 20131101), and 73.7 and 51.1 for the third batch (Batch No. 20131126), respectively. The correlation coefficients between the predicted plasma concentration profiles based on the release profiles of the bio-mimetic DDS and physiological profiles were 0.9366 (Batch No. 20131031), 0.9163 (Batch No. 20131101), 0.9264 (Batch No. 20131126), respectively. Since the pharmacokinetic profile of MT in any kind of animal differs markedly from that of human beings, it is impossible to test the bio-mimetic DDS in animals directly. Therefore, the predicted pharmacokinetic profile based upon the in vitro release kinetics is an acceptable surrogate for the conventional animal test. In this research, a bio-mimetic DDS for replacement of MT was designed with in silico evaluation.

OBJECTIVETo investigate the mechanisms of the oil-in-oil nanoemulsions transport through the gastrointestinal tract and the transport efficiency changed with its different particle size in the lymphatic channels.

METHODThe behavior of nanoemulsions in vivo and their absorption via lymph after oral administration was investigated, with the transport efficiency and absorption pathway of nanoemulsions clarified by lymph duct cannulation in rats.

RESULTIt suggested about 36.8% of puerarin nanoemulsions was transported into systematic circulation via lymph. Nanoparticles with different size absorbed by the lymphatic channels varied as the degree of transportion.

CONCLUSIONThe degree of absorption and particle transport is inversely proportional to the size.

Absorption ; Animals ; Biological Transport ; Emulsions ; Female ; Isoflavones ; administration & dosage ; pharmacokinetics ; Lymph ; metabolism ; Male ; Nanoparticles ; Particle Size ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of Shaoqiduogan (SQDG) on the expression of matrix metalloproteinase 13 (MMP-13) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in carbon tetrachloride (CCl4) induced hepatic fibrosis rats and transforming growth factor beta1 (TGF-beta1) irritated hepatic stellate cells (HSC), and to explore its possible mechanisms.

METHODThe model of chemical hepatic fibrosis induced by CCl4 was prepared. The rats were randomly divided into 5 groups, including normal control group, liver fibrosis model group and SQDG (42. 5, 85, 170 mg x kg(-1)) treated groups. The level of collagen type 1 (C-1) in serum was determined by radioimmunoassay. Masson stain was used to examine the histopathological change. MMP-13 and TIMP-1 ex-pression in liver tissues were assayed by immunohistochemistry. In vitro, effects of SQDG on the expression of MMP-13, TIMP-1 and C-1 in HSC-T6 stimulated by TGF-beta were measured by Western-blot.

RESULTThe results showed that SQDG significantly decreased the elevated level of C-1 in serum of hepatic fibrosis rats induced by CCl4. Pathological examination showed that SQDG could remarkably alleviate the degree of liver fibrogenesis and formation of pseudolobulus. The results of immunohistochemistry demonstrated that SQDG significantly increased MMP-13 expression and decreased TIMP-1 expression in liver tissues. Furthermore, SQDG (20-160 mg x L(-1)) could facilitate MMP-13 expression, inhibit TIMP-1 expression and significantly inhibit the C-I production of HSC stimulated with TGF-beta1 in vitro.

CONCLUSIONThe anti-fibrotic effects of SQDG may be associated with its action of promoting collagen degradation via controlling the levels of MMP-13 and TIMP-1 in liver.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Hepatic Stellate Cells ; drug effects ; enzymology ; metabolism ; Humans ; Liver Cirrhosis ; drug therapy ; enzymology ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism

Aim To investigate the effects of Shaoqiduogan(SQDG)on immunological hepatic fibrosis induced by human albumin in rats as well as its possible mechanisms.Methods The model of immunological hepatic fibrosis induced by human albumin was prepared.The rats were randomly divided into 6 groups,namely normal control group,liver fibrosis model group,SQDG(42.5,85,170 mg·kg~(-1))treated groups and colchicine(0.1 mg·kg~(-1)) treated group.HE staining was used to examine the histopathological change.The activities of transaminase in serum,malondiadehyde(MDA)content,superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)activities,hydroxyproline(Hyp)content in liver homogenate were assayed byspectrophotometry.The levels of hyaluronic acid(HA)and procollagen Ⅲ (PCⅢ)in serum were determined by radioimmunoassay.In vitro,the collagen production of hepatic stellate cell(HSC)-T6 stimulated with transforming growth factor beta1(TGF-β1)was measured with 3H-Proline uptake.Results SQDG had obvious protective effects on human albumin induced hepatic fibrosis in rats.The results showed that the serum ALT and AST decreased by SQDG treatment,but had no significant difference compared with model group.Pathological examination showed that SQDG could remarkably alleviate the hepatic fibrosis.SQDG not only decreased the Hyp content in liver homogenates,but also the elevated level of HA,PCⅢ in serum.SQDG also ameliorated the oxidative stress state of hepatic fibrosis rats,decreased the production of MDA and enhanced the activities of antioxidative enzyme including SOD and GSH-Px.Furthermore,SQDG(20～160 mg·L~(-1))inhibited the collagen production of HSC stimulated with TGF-β1 in vitro.Conclusion sSQDG has protective effect on liver fibrosis rats induced by human albumin.The mechanisms of its anti-fibrotic effects may be associated with its action of ameliorating the oxidative stress in liver,and inhibiting the production of collagen in HSC.

OBJECTIVETo prepare berberine microemulsion, and to investigate its properities and the absorption character in rat intestine in situ.

METHODThe optimum formulation of the blank microemulsion selected by pseudo tertiary phase diagrams and the berberine microemulsion was prepared based on the blank microemulsion. The viscosity, conductance, refraction rate and particle size of berberine microemulsion were surveyed. An in situ rat perfusion method was used to investigate the intestinal absorption of berberine microemulsion. A UV method for determination of berberine in the intestinal flux was established.

RESULTThe viscosity, conductance, refraction rate and particle size of berberine microemulsion were 2.11 cPas, 125.5 microomega, 1.363 and 24.0 nm, respectively. The absorption rate of berberine at the ileum was the best. The absorption of berberine microemulsion at the ileum was significantly higher than that of raw medicine (P < 0.01).

CONCLUSIONThe microemulsion system might improve the absorption of berberine in the intestinal tract.

Absorption ; Administration, Cutaneous ; Animals ; Berberine ; administration & dosage ; pharmacokinetics ; Drug Delivery Systems ; Drug Stability ; Female ; Ileum ; metabolism ; Intestinal Absorption ; drug effects ; Intestines ; metabolism ; Male ; Particle Size ; Rats ; Rats, Wistar ; Skin ; metabolism ; Skin Absorption ; drug effects ; physiology ; Solubility ; Technology, Pharmaceutical ; methods

Objective To optimize the extraction process of total flavonoids in Hypericum perforatum L.. Methods The concentration and volume of alcohol, extraction time and frequency were systematically studied by the total flavonoids as the marker. Results The more effective way to extract total flavones was extraction with 8 times amount of 70% alcohol under reflux for 3 times and 2 h for each time. Conclusion This method obtains more effective components. It is stable and reliable.

Hepatic fibrosis is the hallmark of most chronic liver diseases. Its essense is excessive deposition of extracellular matrix components(ECM) in liver, which occurs due to an imbalance between the production and degradation of matrix. Collagen is the most important part of ECM. Therefore,one important antifibrotic pathway is to effectively inhibit the synthesis of collagen or increase its degradation. The review describes the current progress in collagen metabolism and antifibrotic therapy related to hepatic fibrosis.