Identification of chemical components and components migrating to the blood of Jianpi huazhuo tiaozhi granules / 中国药房

OBJECTIVE To analyze the chemical components and components migrating to the blood of Jianpi huazhuo tiaozhi granules （JHTG）. METHODS SD rats were divided into a control group and a medication group， with 6 rats in each group. The medication group was given JHTG 3 mL. Sixty minutes after medication， the serum samples of the 2 groups were collected， and the chemical components and components migrating to the blood of JHTG were separated by ultra-high performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） and mass spectrometry data were collected. Combined with the overall scheme of UNIFI natural products， based on the 6400 natural product theory mass spectrometry database， the structure was analyzed and confirmed by literature review and reference substance comparison. RESULTS & CONCLUSIONS A total of 130 components were identified from JHTG， including 3 in Codonopsis Radix， 13 in Nelumbinis Folium， 15 in Poria， 5 in Atractylodis Macrocephalae Rhizoma， 9 in Citri Reticulatae Pericarpium， 1 in Coicis Semen， 19 in Alisma Rhizoma， 24 in Salviae Miltiorrhizae Radix et Rhizoma， 7 in Hordei Fructus Germinatus， 24 in Crataegi Fructus， 2 in Amomi Fructus， and 3 in Aucklandiae Radix. In addition， quercetin and quercetin-3-O-β-D-glucopyranoside， kaempferol and citric acid may originate from Nelumbinis Folium or Crataegi Fructus， while oleanolic acid may originate from Poria or Crataegi Fructus. By comparing the reference substances， 8 components were finally determined （pachymic acid， atractylenolide Ⅱ， alisol A， alisol B， alantolactone， bornyl acetate， salvianolic acid A， salvianolic acid C）. A total of 72 prototype components such as quercetin and kaempferol were identified， mainly including flavonoids， terpenoids， lignans and phenolic acids. A total of 11 metabolites such as （NATCM’s Project of High- dehydroanonaine and 16-O-acetylpachymic acid were level Construction of Key TCM Disciplines）identified， mainly terpenoids. Metabolic pathways include phase Ⅰ metabolic reactions such as dehydrogenation and dehydroxylation， and phase Ⅱ metabolic reactions such as methylation and acetylation.

Study on Inhibitory Effects of Berberine on Vascular Endothelial Cells Injury Induced by PM2.5 and Its p38 MAPK Signal Pathway Mechanism / 中国药房

OBJECTIVE:To study the inhibitory effects of berberine on EA.hy926 human umbilical vein endothelial cells (EA. hy926 cells) injury induced by particulates with no more than 2.5 μm air aerodynamic diameter in atmospheric (PM2.5),and its p38 mitogen-activated protein kinase(MAPK)signal pathway mechanism. METHODS:PM2.5 samples were collected and hatched EA.hy926 cells with concentrations of 0(blank control),20,200 and 400 mg/L for 24 h. The survival rate and apoptosis rate of cells,contents of IL-6,TNF-α and MDA,activities of SOD and LDH,protein levels of p-p38 MAPK,Bcl-2 and Bcl-2 associated X protein (Bax) were detected. The above indexes of EA.hy926 cells in blank control group,PM2.5 group (200 mg/L PM2.5), p38 MAPK pathway-specific blocker SB203580 group (20 μmol/L SB203580+200 mg/L PM2.5),berberine low-,medium- and high-concentrations groups(5,10,20 μmol/L berberine+200 mg/L PM2.5)were also determined. RESULTS:Compared with blank control,survival rate of cells,SOD activity and Bcl protein decreased after 200,400 mg/L PM2.5 hatched;apoptosis rate of cells, contents of IL-6,TNF-α and MDA,LDH activity,protein levels of p-p38 MAPK and Bax increased (P<0.05),in concentra-tion-dependent manner. Compared with PM2.5 group,survival rate of cells,SOD activity and Bcl-2 protein increased in berberine medium-,high-concentrations groups and SB203580 group;apoptosis rate of cells,contents of IL-6,TNF-α and MDA,LDH ac-tivity,protein levels of p-p38 MAPK and Bax decreased (P<0.05). CONCLUSIONS:Berberine attenuates PM2.5-induced EA. hy926 cells injury via the inhibition of p38 MAPK pathway.

Tanshinone ⅡA attenuates PM2 .5-induced vascular endothelial cell injury via p38 MAPK signal pathway / 中国病理生理杂志

AIM:To investigate the effect and the mechanism of tanshinone ⅡA in attenuating PM2.5-induced human umbilical vein endothelial EA.hy926 cell injury.METHODS:The samples of fine particulate matter (PM2.5) were collected in Guangzhou and made into suspension.Different concentrations (0, 20, 200 and 400 mg/L) of PM2.5 were added to EA.hy926 cells.The viability and apoptosis of EA.hy926 cells, the protein levels of p-p38 MAPK, Bax and Bcl-2 in the EA.hy926 cells, the contents of interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α) and malonaldehyde ( MDA) , and the activity of superoxide dismutase ( SOD) and lactic dehydrogenase ( LDH) in the EA.hy926 cell culture supernatant were measured by MTT assay, flow cytometry, Western blot, ELISA and colorimetry, respectively.Tanshinone ⅡA at different concentrations (5, 10 and 20 μmol/L) or a specific inhibitor of p38 MAPK pathway, SB203580 (20μmol/L) , was added into the EA.hy926 cells to observe the effect of tanshinone ⅡA.RESULTS:Compared with control group, PM2.5 significantly increased the apoptosis, the contents of IL-6, TNF-αand MDA, the activity of LDH, and the protein levels of p-p38 MAPK and Bax/Bcl-2 ratio, but decreased the viability and SOD activity in the EA.hy926 cells (P<0.05).Compared with PM2.5 group, tanshinone IIA significantly decreased the apoptosis, the contents of IL-6, TNF-αand MDA, the activity of LDH, and the protein levels of p-38 MAPK and Bax/Bcl-2 ratio, but increased the viabil-ity and SOD activity in the EA.hy926 cells (P<0.05).CONCLUSION:Tanshinone ⅡA attenuates PM2.5-induced EA. hy926 cell injury via the inhibition of p38 MAPK pathway.

Allicin prevents EA. hy926 endothelial cell injury induced by PM2. 5 via inhibiting ERK1/2 pathway / 中国药理学通报

Aim To investigate the protective effect of allicin against EA. hy926 endothelial cell injury in-duced by PM2. 5 and the possible mechanism. Meth-ods The samples of fine particulate matter ( PM2. 5 ) were collected and made into suspension. Different concentrations of PM2. 5 ( 20 , 200 , 400 mg · L-1 ) were added to EA. hy926 cell. The viability and apop-tosis of EA. hy926 cell, the protein levels of p-ERK1/2, Bax and Bcl-2 in the EA. hy926 cell, the contents of tumor necrosis factor-α( TNF-α) , interleukin-6 ( IL-6 ) , and malonaldehyde ( MDA ) , the activities of su-peroxide dismutase ( SOD ) and lactic dehydrogenase ( LDH) in the EA. hy926 cell culture supernatant were measured by MTT assay, flow cytometry, Western blot, enzyme-linked immunosorbent assay ( ELISA ) and colorimetry, respectively. Allicin at different con-centrations(5,20,40 mg·L-1 ) or a specific inhibitor of ERK1/2 signaling pathway PD98059 ( 20 μmol · L-1 ) was added into the EA. hy926 cell to observe the effect of allicin. Results Compared with control group, PM2. 5 significantly increased the apoptosis, the contents of TNF-α, IL-6 and MDA, the activity of LDH, the protein levels of p-ERK1/2 and Bax/Bcl-2 ratio, but decreased the viability and SOD activity in the EA. hy926 cell(P<0. 05). Compared with PM2. 5 group, allicin significantly decreased the apoptosis, the contents of TNF-α, IL-6 and MDA, the activity of LDH, the protein levels of p-ERK1/2 and Bax/Bcl-2 ratio, but increased the viability and SOD activity in the EA. hy926 cell ( P <0. 05 ) . Conclusion Allicin displays a significant protective effect against EA. hy926 endothelial cell injury induced by PM2 . 5 and its mechanism may be related to the attenuations of in-flammation and oxidative stress via the inhibition of ERK1/2 pathway.

Effect of Chinese herbal drug-containing serum of Huoxue Bushen prescription on the proliferation of human umbilical vein endothelial cells and on expression of endothelial nitric oxide synthase through p42/44MAPK pathway / 中华老年医学杂志

Objective To observe the drug-containing Huoxue Bushen (HXBS) serum on the proliferation of human umbilical vein endothelial cells (HUVECs) and p42/44MAPK pathways by using human umbilical vein endothelial cells (HUVECs).Methods The proliterative effects of 20％,10％,5％ drug-containing HXBS serum on HUVECs were observed by MTT.Nitric oxide synthase (NOS) activity was detected by endothelial nitric oxide synthase (eNOS) activity kit.The content of nitric oxide (NO) was determined by NO kit.The mRNA expression of eNOS was detected by real-time reverse transcription polymerase chain reaction (RT-PCR).The protein expression of pP42/44 was detected by Western blotting.Results HUVECs proliferation was effectively promoted by drug-containing HXBS serum.The proliferation rate of HUVECs was increased along with the drug-containing HXBS serum concentration increasing after 24 h treatment (10.8 ％,14.7 ％,33.3 ％,t=6.82,6.25,12.16,respectively,P＜0.05 or 0.01).eNOS activity was significantly increased in the drug-containing HXBS serum-treated group as compared with the control group.There was a significant difference in NO content between 20％ drug-containing HXBS serum treated group and control group (P＜0.05).The expression of eNOs mRNA was significantly promoted in 20％ and 10％ drug-containing HXBS serum groups (P ＜ 0.05).P-p42/44 protein level was significantly increased by drug-containing HXBS serum (P ＜ 0.01).Conclusions Conclusions HXBS can promote endothelial cell proliferation and improve endothelial cell NO secretion by increasing the activity and mRNA expression of eNOS through p42/44 signaling pathway,which provides a theoretical basis for improving cardiovascular disease in postmenopausal women.

C3d-M28 enhanced DNA vaccination induced humoral immune response to glycoprotein C of pseudorabies virus / 生物工程学报

We studied the immunogenicity of pseudorabies virus gC DNA vaccination by fusing the murine complement C3d receptor binding domain. First, pseudorabies virus gC gene was linked to four copies of C3d receptor binding domain (M284), and then cloned into the vector pcDNA3.1 to construct the recombinant plasmid sgC-M284. Through the experiment of immunized BALB/c mice, we found that the enzyme linked immunosorbent assay (ELISA) antibody titer for sgC-M284 was 17-fold higher than that for sgC alone, and protective rate of mice was augmented from 25% to 88% after lethal dose PrV (316 LD50) challenge. In addition, the IL-4 levels for sgC-M284 immunization approached that for the pseudorabies virus inactivated vaccine. In conclusion, we demonstrated murine C3d receptor binding domain fusion significantly increased Th2-biased immune response by inducing IL-4 production.