Effect of pre-acupuncture intervention on serum IgE and cutaneous phosphorylated tyrosine-protein kinase expression in rats with urticaria / 针刺研究

OBJECTIVE: To observe the effect of acupuncture on serum IgE level, the degranulation of mast cells, the release of histamine and serotonin, and the expressions of phosphorylated tyrosine-protein kinase Lyn and Syk (p-Lyn, p-Syk) in skin tissue in rats with urticaria, as well as analyze the mechanism of acupuncture in the prevention and the treatment of urticaria. METHODS: SD male rats were randomly divided into normal control, model control, medication and acupuncture groups (n＝10 in each group). The anti-ovalbumin serum was used to establish urticaria model. Rats of the medication group received gastric lavage of Loratadine (0.1 mg/100 g). In the acupuncture group, bilateral "Xuehai" (SP10) and "Quchi" (LI11) were punctured perpendicularly, about 2 to 4 mm in depth, and the needles were retained for 30 min. The treatment was given consecutively for 14 days in the two treatment groups. H．E. staining was adopted to observe the morphological changes of skin tissue, ELISA to determine the total IgE level in serum, the toluidine blue staining to observe the degranulation of mast cells in local skin tissue and the immunohistochemistry to determine the expressions of histamine and serotonin as well as the the expressions of p-Lyn and p-Syk. RESULTS: Compared with the normal control group, the epidermis of the model control group was significantly thickened, the dermis was swollen, the inflammatory infiltration of small vessels was serious and the mast cells were swollen and deformed, with blurred edge and exfoliated granules. Additionally, in the model control group, the serum IgE level was significantly higher (P0.05). CONCLUSION: Acupuncture at LI11 and SP10 is applicable in the treatment of urticaria. This therapy inhibits the type Ⅰ hypersensitivity and the mast cell degranulation, which may be related to the regulation of p-Lyn and p-Syk protein expressions in the locus coeruleus skin tissue.

In vitro metabolism and drug-drug interaction potential of IG-105, a novel antimicrotubule agent / 药学学报

IG-105, N-(2,6-dimethoxypyridine-3-yl)-9-methylcarbazole-3-sulfonamide, a novel antimicrotubule agent, showed potent anticancer activity in a variety of human tumor cells in vitro and in vivo. In order to characterize the metabolism and the possible drug-drug interaction of IG-105, we carried out a series of experiments. Drug metabolizing enzymes involved in IG-105 metabolism were investigated by using pooled human liver microsomes (HLMs) and recombinant cytochrome P450 isoforms (rP450s) respectively. The possible metabolites were analyzed by liquid chromatography-orbitrap-mass spectrometry (LC-Orbitrap-MS). The inhibitory effect of IG-105 on main P450 enzymes was also evaluated. The results showed that IG-105 can be metabolized by a series of rP450s, including CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4 and CYP3A5, with the major contribution enzymes being CYP1A2, CYP2B6, CYP2C19, and CYP3A. Three metabolites (M1-M3) were identified and demethylation was the major phase I metabolic reaction for IG-105. IG-105 moderately inhibited CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A enzyme activities with IC50 values of 6.42, 23.64, 0.39, 1.4, and 3.14 μmol·L-1, respectively. Since the biotransformation of IG-105 involves multiple enzymatic pathways, the compound is less likely to be a victim of a concomitantly used medicine which inhibits activity of one of the CYPs. However, as IG-105 showed medium to strong inhibition on CYP1A2, CYP2D6, CYP3A, and CYP2C19, caution is particularly needed when IG-105 is co-administrated with other anticancer drugs which are mainly metabolized by the above enzymes.

Overexpression of synuclein-gamma confers resistance to antimicrotubule drugs against human hepatoma cells / 药学学报

Liver cancer is one of the most common neoplastic diseases with high mortality in China. Currently, antimicrotubule drugs such as paclitaxel (PTX) and vincristine (VCR), are used as the common agents in the clinical chemotherapy for liver cancer. However, the responses of patients to these drugs vary markedly. Successful identification of intracellular factors influencing liver cancer's sensitivity to antimicrotubule drugs would be of great clinical importance. In this study, by engineering human hepatoma cell HepG2 to overexpress synuclein-gamma (SNCG), we investigated if SNCG is a molecular factor associated with the sensitivity to antimicrotubule drug treatment. Real-time RT-PCR and Western blotting assays showed SNCG was successfully overexpressed in HepG2/ SNCG cells compared with HepG2/Neo cells. The overexpressed SNCG altered the proliferation activity in HepG2 cells, which was 66% higher than that of HepG2/Neo cells through MTT method. The overexpressed SNCG also reduced sensitivity of HepG2 cells to antimicrotubule drugs: after PTX or VCR treatment, the proportion of HepG2/SNCG cells in G2/M arrest was significantly lower than that in HepG2/Neo cells. Correspondingly, HepG2/SNCG cells showed significantly lower mitotic index than HepG2/Neo cells. Meanwhile, HepG2/SNCG cells showed higher resistance to PTX and VCR than HepG2/Neo cells, with resistance index 21 and 15 respectively. Our studies suggested that the overexpression of SNCG could confer resistance to antimicrotubule drugs in hepatoma cells; and it indicated that SNCG may be as a potential response marker for antimicrotubule drugs in liver cancer chemotherapy.