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OBJECTIVE@#To investigate the effects of matrine on antigen presentation of dendritic cells (DCs), and to explore the pharmacological mechanism of matrine on anti-tumor effect.@*METHODS@#Different concentrations (0, 1, 2, 4, 8 and 16 µ g/mL) of matrine were co-cultured with DCs, the harvested DCs were co-cultured with antigens of Lewis lung cancer (LLC) cells, and then DCs and T cells were co-cultured to produce DCs-activated killer (DAK) cells, which have significant tumor-killing activity. The expression of cytokines, mRNA and protein of toll-like receptors (TLRs) in DCs were detected by enzyme linked immunosobent assay, polymerase chain reaction and Western blot, respectively. And the killing effect of DAK were measured by MTT assay.@*RESULTS@#Matrine significantly increased the mRNA expression of TLR7, TLR8, myeloid differentiation factor 88 (MyD88), tumor necrosis factor receptor-associated factor 6 (TRAF-6) and I κ B kinase (IKK), as well as the protein expression of TLR7 and TLR8, and up-regulated the levels of interleukin-12 (IL-12), IL-6 and tumor necrosis factor-α (TNF-α), meanwhile, it also increased the expressions of MHC-II, CD54, CD80 and CD86 in DCs. DCs-activated effector T cells had significant tumor-killing activity. When the concentration of matrine was more than 4 µg/mL, all indices had significant difference (P<0.01 or P<0.05).@*CONCLUSION@#Matrine plays an anti-tumor role by regulating TLRs signal transduction pathway, promoting the secretion of inflammatory cytokines and enhancing immune function.
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Objective:To observe the effect of forsythiaside A on gastrointestinal motility disorder induced by chemotherapy in mice， and explore the mechanism of forsythiaside A regulating gastrointestinal motility. Method:The 60 KM mice were randomly divided into normal group， model group， metoclopramide group （5 mg·kg-1） and forsythiaside A low， medium and high-dose groups （30， 60， 120 mg·kg-1）， 10 for each group， which include half male and half female. The above dose was given once a day for 4 consecutive days， which the intragastric volume was 10 mL·kg-1. One hour after 1rd day administration， equal volume of saline was intraperitoneally injected to the normal group， 2 mg·kg-1 cisplatin was intraperitoneally injected to the other groups with daily for 4 consecutive days. Observing the effects of forsythiaside A on gastric emptying and small intestinal propulsion on mice models， serum gastrin （GAS） and somatostatin （SS）， motilin （MTL）， vasoactive intestinal peptide （VIP） levels were examined by enzyme-linked immunosorbent assay （ELISA）. Activities of acetylcholinesterase （AChE） and total nitric oxide synthase （tNOS） in gastric antrum and ileum were detected by ELISA. The expression of AChE and inducible nitric oxide synthase （iNOS） in gastric antrum and ileum were detected by Western blot. Result:Compared with normal group， the gastric retention rate and small intestinal propulsion rate of the model group were significantly increased （P<0.01）， serum levels of MTL， GAS， SS and VIP， the AChE activity in the homogenate of ileum in the model group were significantly reduced （P<0.05，P<0.01）， while the tNOS activities in gastric antrum and ileum were significantly increased （P<0.05，P<0.01）. Protein expression of AChE in gastric antrum and ileum were significantly decreased （P<0.05）， and the expression level of iNOS protein was significantly increased in the model group （P<0.05）. Compared with model group， different doses of forsythiaside A can reduce the gastric residual rate and small intestinal propulsion rate of mice to varying degrees. Meanwhile forsythiaside A can increase the serum levels of MTL， GAS， SS， and VIP， and the AChE activity and protein expression levels in gastric antrum and ileum tissues were also increased， while tNOS activity and iNOS protein expression were decreased in gastric antrum and ileum （P<0.05，P<0.01）. Conclusion:Forsythiaside A can significantly ameliorate the delayed gastric emptying and small intestine hyperfunction induced by cisplatin in mice. Its mechanism to ameliorate gastrointestinal dysfunction caused by chemotherapy is related to the regulation of gastrointestinal AChE and NOS activity in gastric antrum and ileum and the regulation of gastrointestinal hormone levels.
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Objective: To establish HPLC fingerprint of Notopterygii Rhizoma et Radix with different commercial specifications and to provide the basis for the division of commercial grades and the quality control of Notopterygii Rhizoma et Radix on the market. Method: The market investigation and literature research were used to understand the existing situation of Notopterygii Rhizoma et Radix goods.Notopterygii Rhizoma et Radix goods were divided into three commercial specifications according to the source and appearance,such as Canqiang,Tiaoqiang and Datouqiang.Fingerprint of Notopterygii Rhizoma et Radix with different commercial specifications was established by HPLC-PDA,the mobile phase was consisted of acetonitrile-0.3% acetic acid in a gradient elution mode.Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine(version of 2004A) was used to confirm the common peaks and evaluate the similarity.SPSS 19.0 statistical software was used to make principal component analysis(PCA) for HPLC fingerprint pattern. Result: The common mode of fingerprint for Canqiang,Tiaoqiang and Datouqiang were established separately.A total of 22 common peaks were marked in Canqiang,23 common peaks were marked in Tiaoqiang,29 common peaks were marked in Datouqiang.The result of similarity evaluation and PCA showed that the quality of Notopterygii Rhizoma et Radix with the same commercial specification was stable.There were great differences in chemical compositions and their contents among Notopterygii Rhizoma et Radix with different commercial specifications. Conclusion: The fingerprint method can well distinguish commercial specifications of Notopterygii Rhizoma et Radix,and it can provide the basis for the division of commercial grades and the quality control of Notopterygii Rhizoma et Radix.
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Objective: To observe effect of Mori Folium-containing serum on glucose consumption and cell activity of fat cell line 3T3-L1 insulin resistance (IR) model, in order to screen out the optimal concentration of drug-containing serum, detect effect of Mori Folium on the content of inflammatory factors, and explore the possible mechanism. Method: 3T3-L1 preadipocytes in logarithmic growth phase were selected, and induced with 10 mg·L-1 insulin (Ins), 0.25 mmol·L-1 dexamethasone (DEX) and 0.5 mmol·L-1 3-isobutyl-methylxanthine(IBMX) for 48 h and then with 10 mg·L-1 Ins for 48 h. After the cells were differentiated into mature adipocytes, they were induced with 1 μmol·L-1 DEX for 96 h to establish IR model. Glucose content in the supernatant of cells was detected by glucose oxidase after serum containing Mori Folium cultured for 12,24,36, 72 h. Methyl-thiazdyl-tetrazolium(MTT) was used to detect the effect of serum containing Mori Folium on IR cells activity. The content of tumor necrosis factor-α(TNF-α) was determined by enzyme-linked immunosorbent assay (ELISA). Meanwhile, the effects of inflammatory factors on the expressions of insulin signaling pathway proteins insulin receptor (InsR), insulin receptor substrate (IRS), p-IRS1 and glucose transporter 4 (GLUT4) were determined by Western blot. Result: Serum containing Mori Folium could significantly increase the glucose consumption rate and cell activity of IR cells (Pα (PPPConclusion: Mori Folium can significantly improve IR status of 3T3-L1 cells, and its mechanism may be related to inhibiting TNF-α and promoting the expressions of insulin signaling pathway proteins.
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This study is to establish a qualitative method for rapid identification of bile acids in Suis Fellis Pulvis based on UHPLC-LTQ-Orbitrap-MS technology,and an HPLC-ELSD internal standard method for the quantitative determination of two glycine-conjugated BAs in Suis Fellis Pulvis.The chromatographic separation of the UHPLC-LTQ-Orbitrap-MS qualitative analysis was achieved on a Waters Acquity UPLC HSS T_3column(2.1 mm×100 mm,1.8μm),with 0.2%formic acid aqueous solution(A)-acetonitrile(B)as mobile phase ingradient elution.Electrospray ionization(ESI)source was applied and operated in negative ion mode.Quantitative analysis was performed at 30℃on a Diamonsil-C_(18)column(4.6 mm×250 mm,5μm).The mobile phase consisted of 0.2%formic acid solution and acetonitrile with gradient elution and the flow rate was 1.0 m L·min~(-1).An ELSD was used with a nitrogen flow-rate of1.4 L·min~(-1)at a drift tube temperature of 60℃and the gain was 1.A total of 14 bile acids in Suis Fellis Pulvis were characterized based on the accurate mass measurements,fragmentation patterns,chromatographic retention times,and reference materials.For the quantitative analysis method,the glycohyodeoxycholic acid and glycochenodeoxycholic acid had good linear relationship in the range of26.52-265.20 mg·L~(-1)(r=0.999 8)and 19.84-198.40 mg·L~(-1)(r=0.999 1),respectively.The average recoveries(n=6)were104.1%and 103.1%,and the RSD were 2.0%and 2.4%.The UHPLC-LTQ-Orbitrap-MS technology provides a fast and efficient qualitative analysis method for identification of bile acids in Suis Fellis Pulvis.The HPLC-ELSD internal standard method is accurate and reliable,which has reference value for the quality control of Suis Fellis Pulvis.
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Animals , Cholic Acids , Chromatography, High Pressure Liquid , Quality Control , SwineABSTRACT
Objective To investigate the effects of Jinqi Zhizhu Decoction (JQZZD) on gastric motility of GK rats with diabetic gastroparesis (DGP); To discuss its mechanism of action. Methods 12 out of 80 male GK rats were randomly selected as normal group, and others were induced to be DGP model by irregular feeding with high sugar and high fat diet for 8 weeks. The model rats were randomly divided into model group, positive medicine group, JQZZD low-, medium-, and high-dose groups. All rats were given continuous intragastric administration for 12 weeks. The residual rate of pigment in stomach was detected and calculated. The concentration of DA, ACh and 5-HT in plasma were detected by ELISA. The morphological changes of gastric antrum were observed by HE staining. The ultrastructure of mucous layer of gastric antrum was observed by transmission electron microscope. The protein expressions of Rho A, ROCK1 and ROCK2 were detected by Western blot. Results Compared with normal group, the residual rate of pigment in model group increased, while DA, ACh and 5-HT, relative amount of protein expressions of Rho A, ROCK1 and ROCK2 decreased (P＜0.01). Compared with model group, the residual rate of pigment in positive medicine group and JQZZD high-dose group decreased, while DA, ACh and 5-HT, relative amount of protein expressions of Rho A, ROCK1 and ROCK2 increased (P＜0.05, P＜0.01), and morphological changes and ultrastructure of mucous layer obviously improved. There was no significant differences in the residual rate of pigment, DA content and protein expressions of Rho A, ROCK1 and ROCK2 between positive medicine group and JQZZD high-dose group (P＞0.05). Compared with JQZZD low-dose group, the residual rate of pigment in JQZZD high-dose group decreased, while DA, ACh and 5-HT, relative amount of protein expressions of Rho A, ROCK1 and ROCK2 increased (P＜0.05, P＜0.01). Conclusion JQZZD may improve gastric motility, promote gastric emptying, and improve and repair damage of mucous membrane in the gastric antrum. All of these may be related to regulating protein expressions of Rho A/ROCK signal pathway.