Study On The Screening Of Anti - Inflammatory And Analgesic Active Sites And Its Mechanism

Cowherb Seeds are the dried ripe seeds of Vaccaria segetalis (Neck.) Garcke, which is belonging to the family of Caryophyllaceae. Bitter in taste and mild in medicine property, it is often used in the treatment of with the diseases associated with liver and stomach, it also has the capacity to promote milk secretion and activate blood flow, and it is often used in the treatment of breast infections and amenorrhea. In resent years, it has been used for amenorrhea, dysmenorrhes and so on in clinic. Cowherb Seeds contain a variety of ring glycosides, cyclic peptide and flavonoid glycosides, such as apigenin-6-C-arabinose-glucoside, Cowherb Seeds cyclic peptide A and Cowherb Seeds flavonoid glycosides. Modern pharmacological studies show that Cowherb Seeds can not only promote the lactation and inhibite the angiogenesis and has anti-oxidation activity; its decoction also has good anti-inflammatory and analgesic effects. However, the mechanism of Cowherb Seeds’ anti-inflammatory and analgesic effection is unclear. In this study Cowherb Seeds extract was separated into different polar fractions using the method of plant chemistry, to screen active fractions from Cowherb Seeds. The anti-inflammatory and analgesic effects of the active fractions were tested by using pharmacological methods in vivo. The molecular mechanisms of the active fractions were investigated by using biochemical and molecular biology methods. The experimental researches were performanced as following:1The anti-inflammatory effects of the Cowherb Seeds and fried Cowherb Seeds extracts were detected using the xylene-induced ear swelling model in mice. Except the aether extracts from the Cowherb Seeds and fried Cowherb Seeds, the acetoacetate, n-Butanol and water extracts from the Cowherb Seeds and fried Cowherb Seeds significantly reduced the xylene-induced ear swelling in mice in300mg/kg dose. The fried Cowherb Seeds extracts showed more potent anti-inflammatory effects than the Cowherb Seeds extracts in acute inflammation model in mice. The acetoacetate extracts showed the best anti-inflammatory effects in different Cowherb Seeds extracts. So, the anti-inflammatory components should be different in Cowherb Seeds and fried Cowherb Seeds.2The anti-inflammatory effects of the fried Cowherb Seeds extracts were detected using the cotton-induced granuloma model in mice. The results show that the acetoacetate, n-Butanol and water extracts from the fried Cowherb Seeds significantly reduced the cotton-induced granuloma in a dose-dependent manner. So, oral administration of the three extracts also produced significant anti-inflammatory effects in chronic inflammation model in mice. And the anti-inflammatory effect of the acetoacetate extract was much better than the other two extracts.3The analgesic effects of the fried Cowherb Seeds extracts were detected using the acetic acid induced abdominal writhing model in mice. The results show that the three extracts from the fried Cowherb Seeds significantly reduced the acid-induced writhing number in mice. The analgesic effect of n-butanol extract was much better than the other two extracts.4The analgesic effects of the fried Cowherb Seeds extracts were detected using the hot-plate test model in mice. The three extracts from the fried Cowherb Seeds significantly reduced the heat-induced licking numbers in mice. The analgesic effect of n-butanol extract was much better than the other two extracts. The results above were consistent with the results of the acetic acid induced abdominal writhing model in mice.5In order to clarify the mechanism involved in the anti-inflammatory activity of the fried Cowherb Seeds extracts, their effects on malondialdehyde (MDA) and tumor necrosis factor-a (TNF-a) in mice serum and nitric oxide (NO) production in liver tissue were examined. The results showed that the acetoacetate, n-Butanol and water extracts from the fried Cowherb Seeds significantly decreased the MDA and TNF-a level in serum of cotton-induced granuloma mice in dose-dependent manner. Furthermore, they inhibited NO, TNOS and iNOS production in liver tissue of cotton-induced granuloma mice in a dose-dependent manner. These results imply that the anti-inflammatory mechanism of the fried Cowherb Seeds extracts may be explained at least in part, by the inhibition of MDA and TNF-a and NO production.6The conditions for the extraction of the total flavonoids from the fried Cowherb Seeds were optimized using ultrasonic extraction method. The results showed that the better conditions were3times extraction with10times of50%ethanol at60℃,30min for every time.