Main components in butyl alcohol extract of Baitouweng Decoction inhibited neutrophil chemotaxis / 中国中药杂志

The present study aims to investigate the effects of the main components(aesculin, berberine hydrochloride, and anemoside B4) in the butyl alcohol extract of Baitouweng Decoction(BAEB) on the chemotaxis of neutrophils induced by dimethyl sulfoxide(DMSO). HL60 cells were cultivated in RPMI-1640 complete medium, and transferred into a 6-well plate(2 × 10~5 per mL) with 4 mL in each well, followed by incubation with DMSO at 1.3% for five days. The morphologic changes of cells were observed under an inverted microscope. The CD11 b expression after DMSO induction was analyzed by flow cytometry. The effects of aesculin, berberine hydrochloride, and anemoside B4 on the cell proliferation and migration were detected by CCK8 assay and Transwell assay, respectively. The effects of the main components on the production and polarization of F-actin protein were also examined by flow cytometry and laser confocal microscopy. PI3 K/Akt signaling pathway was checked by Western blot. As revealed by the results, neutrophil-like HL60 cells were observed after DMSO induction. The CD11 b expression in these cells increased significantly as indicated by the flow cytometry. Additionally, 100 μg·mL~(-1) aesculin, 8 μg·mL~(-1) berberine hydrochloride, and 80 μg·mL~(-1) anemoside B4 were potent in inhibiting the migration of neutrophils and reducing F-actin expression. Berberine hydrochloride was verified to be capable of diminishing phosphorylated PI3 K/Akt protein expression. The findings indicate that aesculin, anemoside B4, and especially berberine hydrochloride in the BAEB can inhibit the chemotaxis of neutrophils, which is possibly achieved by the inhibition of F-actin and PI3 K/Akt signaling pathway.

[Effect of butyl alcohol extract of Baitouweng Decoction on epithelial barrier of vaginal mucosa in mice with vulvovaginal candidiasis].

To investigate the effect of butyl alcohol extract of Baitouweng Decoction(BAEB) on the epithelial barrier of vaginal mucosa in mice with vulvovaginal candidiasis(VVC). Seventy-two female SPF Kunming mice were randomly divided into blank group, VVC model group, fluconazole group, and BAEB treatment groups(high, middle and low dose groups). Estradiol benzoate was injected subcutaneously qd alt, and Candida albicans(2×10~6 CFU·mL~(-1)) was inoculated into the vagina of mice during the pseudo estrus period for 7 days to construct a VVC model, followed by drug treatment for 7 days. Gram staining was used to observe the morphology of C. albicans in the vaginal secretions of mice; the amount of fungal load on the vaginal mucosa of mice was detected on agar plate; the pathological status of murine vaginal mucosa was observed by hematoxylin-eosin staining(HE); the integrity of mice vaginal mucosal epithelial barrier was observed by Masson's trichrome staining(MT), HE and periodic acid-schiff staining(PAS). Mucin-1 and mucin-4 protein expression levels of vaginal mucosal epithelial cells in mice were detected by immunohistochemistry; mucin-1 and mucin-4 protein expression levels on mucosal epithelial cells at 0 d, 3 d, and 7 d were determined by Western blot. The results showed that, in VVC model group, there were a large number of C. albicans hyphae and higher fungal load in vagina, within complete mucosal structure, cornified layer shed off, and the protein expression levels of mucin-1 and mucin-4 were significantly increased. After BAEB treatment, the hyphae in the vagina decreased; the fungal load decreased; the vaginal mucosal tissue damages were improved; the epithelial barrier was repaired, and mucin-1 and mucin-4 protein expression levels were down-regulated. The above results indicated that BAEB may play a role in the treatment of VVC by remodeling the integrity of the vaginal mucosal epithelial barrier.