Deaggregators inhibit TNF-α-induced leukocyte adhesion in vitro by breaking up hydrophobic lipophilic interactions.

AIM: Deaggregators (deAgrs) are nontoxic organic molecules that possess the ability to deaggregate simple aggregates formed by hydrophobic lipophilic interactions (HLI). Since HLI-driven organic molecule aggregates may induce leukocyte adhesion, we investigated the influence of deAgrs on TNF-α-mediated leukocyte adhesion in vitro. METHODS: For adhesion studies, vascular endothelial cells or smooth muscle cells monolayers were treated with TNF-α (10 µg/L) and deAgrs for 24 h, followed by addition of monocytes or neutrophils suspension. The non-adherent leukocytes were rinsed, and the number of attached leukocytes was measured using an ELISA plate reader. Simultaneously, fluorescence probes Np-12 and Np-Ch were used to measure the deaggregating efficiencies of these deAgrs. RESULTS: Among the nine deAgrs tested,eight significantly reduced the cell adhesion rates with the order of efficiencies: 260 > 160 > 568 > ZPMOP > R68 > 640 > TB6PMOP > CNS, but TBHQ had no effect. The deAgrs for deaggregating an aggregated probe (Np-12 or Np-Ch) exhibited a similar order of efficiencies: 260 > 160 > 568 > ZPMOP > R68 > 640 > TB6PMOP > CNS > 12-AA > 11-AA > TBHQ. Spearman correlation coefficient analyses indicated that the adherent rates of leukocytes to endothelial cells or smooth muscle cells treated with deAgrs had significantly negative correlation to their deaggregating abilities. CONCLUSION: DeAgrs effectively inhibit TNF-α-mediated leukocyte adhesion in vitro by breaking up hydrophobic lipophilic interactions, thus may be further tested for blocking atherogenesis.

[Studies on anti-tumor and enhancing immunity activity of toad coat].

OBJECTIVE: To study the anti-tumor and immunity activity of toad coat (Chantui), which is a new officinal part of Bufo bufo gargarizans. METHOD: The tumor weight of S180, H22, Lewis lung cancer cell inoculated in mice were compared between the groups of mice, fed with toad coat, and those which were not (control group). The average longevity of the mice with HCA fed with toad coat was also compared with the control group. The T lymphocyte transformation and NK cell killing activity were tested and compared with the control group. The condition of the mice which were fed with great dosage of Chantui (16 g x kg(-1) x d(-1)) was observed. RESULT: The tumor weight was remarkably reduced in the groups which were fed with toad coat compared with the control group. Tests show that toad coat can raise the activity of both T lymphocyte and NK cell. There was no obvious side-effect when the mice were fed with great dosage of toad coat. CONCLUSION: The results show that toad coat has a strong inhibitory activity against tumors inoculated in mice and a strong enhancement of immune activity, so it could be viewed as a new valuable safe medicinal source.

[Structural characterization of a glucan isolated from Gastrodia elata].

OBJECTIVE: To study a glucan (GB II) isolated from Gastrodia elata. METHODS: The glucan was obtained with water extraction, alcohol precipitate, DEAE-Sepharose Fast Flow column and Sepharose CL-6B column chromatography; sugar composition analysis, IR and NMR were used to determine the structural feature. RESULTS: The molecular weight of the glucan was 4 300 dalton estimated by HPGPC; it contained 27 glucose residues, which mainly composed of alpha-D-(1-->4)-glucose with little glucuronic acid and branch O-6 points. CONCLUSION: The glucan was a new glucan for the first report from Gastrodia elata.

Optimization of extraction and purification of active fractions from Schisandra chinensis (Turcz.) and its osteoblastic proliferation stimulating activity.

Extraction and purification conditions of lignans from the fruits and seeds of Schisandra chinensis (Turcz.) were investigated through an orthogonal design of L(9)(3(4)) assay and macroporous resin technology. The extraction was optimized using 95% ethanol. For purification, the extract was dissolved in 30% ethanol, then adsorbed on a AB-8 macroporous resin and eluted with 30% ethanol and 70% ethanol successively, the latter resulting in a residue containing 65.2% of lignans. By HPLC analysis schisandrin, deoxyschisandrin and gamma-schisandrin were quantitatively determined. UMR 106 cells were used to examine the stimulatory activity of the lignans on osteoblasts in vitro. The lignans stimulated the proliferation of and the activity of alkaline phosphatase in the osteoblasts indicating their potential activity against osteoporosis.

[Determination of stachydrine and leonurine in Herba Leonuri by ion- pair reversed-phase high-performance liquid chromatography].

OBJECTIVE: To establish a high-performance liquid chromatography (HPLC) for determining stachydrine and leonurine contents in the crude drug of Yimu Cao, a Chinese motherwort herb(Herba Leonuri, the aerial part of Leonurus japonicus). METHODS: The sample was obtained from the crude drug by ultrasonic extraction with ethanol after decolorization with ethyl acetate. YMC-Park CN column (250.0 mm x 4.6 mm) was used with the mobile phase for stachydrine of 0.00125 mol/L SDS:C2H5CN (90:10) at the flow rate of 1.5 ml/min with the wavelength for detection of 201.7 nm, and that for leonurine of 0.00125 mol/L SDS (containing 0.05% HClO4):CH4OH(90:10) at the flow rate of 1 ml/min with the wavelength for detection of 282 nm. The sensitivity was 0.1 AUFs and the column temperature was 20 degrees Celsius;. RESULTS: With the injection amount (mg) as the abscissa and the peak area as the ordinate, the regression equation of the calibration curve for stachydrine was Y=1,187.542 3X-168.9822, and that for leonurine was Y=5,202.654X-221.141 (r=0.9998) with the linearity scope of 2.5-12.5 mg, detective limit of 0.15 mg, and recovery of (99.03+/-2.744) %. The analysis of extracted drug samples from 18 regions indicated that the alkaloids contents in Yimu Cao varied significantly, i.e. stachydrine within a range of 0.1%-0.2% while leonurine content, which was much lower, within 0.01%-0.05%. In general, the alkaloid contents were higher in the drug produced in northern China than in those produced in southern China. L. japonicus and the variant L. japonicus albiflorus had total alkaloid contents around 0.3%, which was higher than the contents of other species. CONCLUSION: As a convenient and feasible means for determining the alkaloid contents in Yimu Cao, HPLC produces accurate and reliable results and can be more effective than the formerly used methods.

Deaggregation behavior of the phenol gemini derivatives.

The phenol gemini compounds were synthesized by rebuilding the substituents and lateral chain on the phenyl ring. Two parts of the molecules were connected by a spacer of hydrocarbon chain via ester groups. The deaggregation behaviors of target molecules were investigated by studying coaggregation behaviors, evaluating the deaggregating abilities in terms of fluorescence measurements, the measurements of their aggregation number and the effects on restraining the cell adhesion. In present cases, all of these experiments give the same results; that is, in these molecules, the longer branched methyl hydrocarbon chain and the combination of methoxy with hydroxyl groups on the phenyl ring possess the higher deaggregating abilities in the studied situation. The molecules with two hydrocarbon chains, i.e., the phenol gemini compounds, are more effective to deaggregate the aggregated probes than the molecules with one hydrocarbon chain. To find an effective deaggregator might be helpful to design the medicine for the cure of arteriosclerosis.