Effects of aqueous extracts of Taraxacum Officinale on expression of tumor necrosis factor-alpha and intracellular adhesion molecule 1 in LPS-stimulated RMMVECs.

BACKGROUND: Mastitis gives rise to big financial burden to farm industry (mainly dairy production) and public health. Its incidence is currently high and therefore, highly effective treatments for therapy, especially with natural products are required. Taraxacum officinale has been reported to use for anti-inflammation. However, its effect on endothelium during mastitis has not been reported. METHODS: We firstly established inflammation experimental model of rat mammary microvascular endothelial cells (RMMVECs). We evaluated the effects of dandelion leaf aqueous extracts (DAE) on LPS-induced production of inflammatory mediators in RMMVECs by enzyme-linked immunosorbent assay and Western blot. We treated RMMVECs with 1 µg/ml LPS for 4 h and then incubated with 10, 100 and 200 µg/mL DAE for 4, 8, 12 and 24 h. The expression (mRNA and protein level) of targets (tumor necrosis factor-alpha (TNF- α) and Intracellular Adhesion Molecule 1 (ICAM1) was analyzed by employing real-time PCR and Western blots. The in vivo anti-inflammatory effect of DAE on mastitis within an Staphylococcus aureus-induced mouse model was also determined. RESULTS: The obtained results showed that dandelion extracts at the concentration of 100 and 200 µg/mL could significantly inhibit both TNF-α and ICAM-1 expression in all time points checked while 10 µg/mL of dandelion only suppress both expression at 8 and 12 h post-treatment. The in vivo tests showed that the DAE inhibited the expression of TNF-α and ICAM-1 in a time-dependent manner. CONCLUSIONS: All results suggest that the endothelium may use as as a possible target of dandelion for anti-inflammation.

Chinese herbal medicinal ingredients inhibit secretion of IL-6, IL-8, E-selectin and TXB2 in LPS-induced rat intestinal microvascular endothelial cells.

The aim of the research was to investigate the anti-inflammatory mechanism of Pulsatillae Decoction (PD), the levels of interleukin (IL)-6, IL-8, E-selectin, and thromboxane B(2) (TXB(2)) secreted by cultured rat intestinal microvascular endothelial cells (RIMECs) were determined after treatment with its active ingredients, namely anemoside B4, anemonin, berberine, jatrorrhizine, palmatine, aesculin, and esculetin. RIMECs were challenged with 1 microg/mL lipopolysaccharide (LPS) for 3 h, and then treated with each of the seven ingredients at three concentrations (1, 5 and 10 microg/mL) for 24 h. The results revealed that anemonin, aesculin and esculetin inhibited the production of IL-6, aesculin and esculetin inhibited the secretion of IL-8, anemoside B4, berberine and jatrorrhizine downregulated E-selectin expression, anemonin, berberine, jatrorrhizine and palmatine decreased the content of TXB(2). All these changes were significant. Taken together, the data suggest that all seven active ingredients of PD can effectively reduce inflammatory response, thus relieving intestinal dysfunction via multiple pathways.

Pulsatilla decoction and its active ingredients inhibit secretion of NO, ET-1, TNF-alpha, and IL-1 alpha in LPS-induced rat intestinal microvascular endothelial cells.

To investigate the pharmacological mechanism of the traditional Chinese medicine, Pulsatilla decoction (PD), the levels of nitric oxide (NO), endothelin-1 (ET-1), tumor necrosis factor-alpha (TNF-alpha), and interleukin-1 alpha (IL-1 alpha) secreted by cultured rat intestinal microvascular endothelial cells (RIMECs) were determined after treatment with PD and its seven active ingredients, namely anemoside B(4), anemonin, berberine, jatrorrhizine, palmatine, aesculin, and esculetin. RIMECs were challenged with lipopolysaccharide (LPS) at 1 microg ml(-1) for 3 h and then treated with PD at 1, 5, and 10 mg ml(-1) and its seven ingredients at 1, 5, and 10 microg ml(-1) for 21 h, respectively. The results revealed that PD, anemonin, berberine, and esculetin inhibited the production of NO; PD, anemonin, and esculetin inhibited the secretion of ET-1; PD, anemoside B(4), berberine, jatrorrhizine, and aesculin downregulated TNF-alpha expression; PD, anemoside B(4), berberine, and palmatine decreased the content of IL-1 alpha. It showed that PD and its active ingredients could significantly inhibit the secretion of NO, ET-1, TNF-alpha, and IL-1 alpha in LPS-induced RIMECs and suggested they would reduce inflammatory response via these cytokines.

[Effect of berberine on nitric oxide release of rat intestinal mucous microvascular endothelial cells].

OBJECTIVE: To study the effect of berberine on the release of nitric oxide (NO) by rat intestinal mucous microvascular endothelial cells (RIMECs) cultured in vitro for exploring the pharmacological mechanism of berberine in treating intestinal disease. METHODS: Cultured RIMECs in vitro were identified adopting immunofluorescent stain with factor VIII-related antigen. NO level in the supernatant of normal cell culture or cell culture treated with different concentrations of berberine was detected with colorimetry at the 3rd, 6th, 9th and 12th h and compared. RESULTS: Compared with that in the normal control group, NO level increased more obviously in the berberine treated groups, and the best effect was shown in the 5 microg/mL berberine treated group. CONCLUSION: One of the important pharmacological mechanisms of berberine might be through promoting the endogenous NO release to induce endothelium-dependent dilatation of microvascular in intestinal mucosa, thus to improve the local microcirculation of intestine.

Effect of anemonin on NO, ET-1 and ICAM-1 production in rat intestinal microvascular endothelial cells.

Anemonin (the dilactone of cyclobutane-1, 2-diol-1, 2-diacrylic acid) was isolated from the root of Pulsatilla chinensis Regel. Pulsatilla chinensis Regel has been used in the treatment of enteritis in China for years. However, only little was known about the mechanism underlying its anti-inflammatory effects. We investigated the effect of anemonin on the release of nitric oxide (NO), endothelin-1 (ET-1) and soluble intercellular adhesion molecule-1 (sICAM-1) induced by lipopolysaccharide (LPS) in primary cultures of rat intestinal microvascular endothelial cells (RIMECs). RIMECs were challenged with 1 microg/ml LPS with or without the presence of various concentrations of anemonin (1, 5 and 10 microg/ml). Anemonin significantly inhibited the production of NO and ET-1 induced by LPS at a concentration of 5 microg/ml and at 10 microg/ml anemonin down-regulated LPS-induced sICAM-1 expression. Anemonin itself had no effect on either factor. These findings suggest that anemonin may exert some beneficial therapeutic action in intestinal inflammation, at least in part by inhibiting the production of NO, ET-1 and ICAM-1 in RIMECs and thus preventing intestinal microvascular dysfunction.

[Effect of electroacupuncture on expression of IFN-gamma-like immunoreactive substance in spinal cord of the rat ].

OBJECTIVE: To observe effect of electroacupuncture at "Zusanli" (ST 36) on expression of interferon-gamma (IFN-gamma)-like immunoreactive substance in spinal cord of the rat and to probe the mechanism. METHODS: The IFN-gamma-like immunoreactive positive cell number in spinal cord of the rat was investigated with immunohistochemical SP method and microscopy. RESULTS: After electroacupuncture at "Zusanli" (ST 36), IFN-gamma-like immunoreactive positive cell number in the spinal cord of the rat with electroacupuncture plus immunosuppression did not significantly change (P > 0.05). The number of positive cells in the dorsal horn in the rats with immunosuppression was significantly less than that in the normal control group (P < 0.05) with no significant change in other parts and with no significant difference between the electroacupuncture group and the control group (P > 0.05). CONCLUSION: Electroacupuncture at "Zusanli" (ST 36) can increase expression of IFN-gamma-like immunoreactive substance in spinal cord of the rat, and acupuncture activates the nerve-immunoregulative network possibly by IFN-gamma as medium.