| Objective: The onset and progression of periodontitis have been shown to be associated with both quantitative and qualitative changes of Gram-negative bacteria. Bacterial endotoxins are unique glycolipids present in the outer cell membrane of all Gram-negative bacterium. Endotoxins is known to initiate the pathophysiological changes that often accompany Gram-negative bacterial infections, its cytotoxicity and immunologic competence make important roles in the progression of periodontitis. Therefore, anti-endotoxin is a key point to prevent it from continuing development. Chinese medicinal herb is paid close attention by scholars of world because of tiny ill effect, source economy and so on. So far, many researches have shown that Chinese medicinal herb can prevent and cure oral disease, and find many of them possess preventive and therapeutic effect on periodontal disease. The researches on mechanisms of cureing periodontitis with Chinese medicinal herb are occupying approach stage. Material ground of pharmacological action is chemical composition.many of traditional Chinese medicine have been found to posses effect of anti-endotoxin, but the researches on chemical composition are less. Scutellaria is an origanum,which is recorded to possess fairly wide antibacterial spectrum in 《Chinese orthodox medicine》.Among its effective ingredient, baicalin has the strong pharmacological action. We adopted limulus test in the study to observe the degrading effect of baicalin on endotoxin, to seek a medicine which prevent and cure periodontitis efficiently to provide base of theory. Methods: Baicalin 10 ㎎was weighed and put in the measuring flask of 100mL, prepared to become reserve of PH 7.60, concentration 100ug·mL -1.By adopting successive double dilution the reserve was diluted to become baicalin solution of 100ug·mL-1,50ug·mL-1,25ug·mL-1,12.5ug·mL-1. Limulus amebocyte lysate and specific limulus amebocyte lysate were dissolved by 0.15mL depyrogenation water, added 0.15mL baicalin solution of 100ug·mL-1,50ug·mL-1,25ug·mL-1 ,12.5ug·mL-1, respectively. Limulus test was applied to observe whether baicalin impact gelation of limulus amebocyte lysate. Endotoxin standard preparation(10EU) was diluted to become 1EU·mL-1. By adopting successive double dilution the reserve was diluted to become 0.5mL baicalin solution of 12.5ug·mL-1 ,6.25ug·mL-1 ,3.125ug·mL-1 ,1.562ug·mL-1 ,0.781ug·mL-1,were put in non-pyretogenous tubes,respectively.1EU·mL-1 endotoxin standard preparation 0.5mL were added in the tubes, too. The tubes were incubated at (37±1)℃for 15min,30min,60min. Limulus test was applied to test the degrading effect of baicalin solution on endotoxin. Meanwhile positive control,negative control and baicalin blank were done. Results: 1 ,Baicalin solution of concentration ≥100ug·mL-1 induced gelation of limulus amebocyte lysate.It did not induce gelation of limulus amebocyte lysate as its concentration≤50ug·mL-1 . 2,The degrading effect of 12.5ug·mL-1,6.25ug·mL-1 baicalin solution on 1EU·mL-1 endotoxin were fairly strong, endotoxin was degraded completely in 15min. 3,The degrading effect of 3.125ug·mL-1 baicalin solution on 1EU·mL-1 endotoxin:endotoxin value was (0.1555±0.0028)EU·mL-1 after the mixture had been incubated for 15min. Positive control was (1.0366±0.1779)EU·mL-1.These differences were statistically significant(P<0.01). Endotoxin was degraded completely in 30min. 4,The degrading effect of 1.562ug·mL-1 baicalin solution on 1EU·mL-1 endotoxin: after the mixture had been incubated for 15min and 30min, endotoxin values were (0.2121±0.0049)EU·mL-1 ,(0.1640±0.0025)EU·mL-1 ,respectively. Positive control was (1.0277±0.0191)EU·mL-1. These differences among them were statistically significant(P<0.01). Endotoxin was degraded completely in 60min. 5,The degrading effect of 0.781ug·mL-1 baicalin solution on1EU·mL-1 endotoxin: after the mixture had been incubated for 15min , 30m... |
PDF Full Text Request