| Object To study on extraction and priliminary test of chemicalconstituents from Ajuga decumbens, then to screen of the active part of Ajugadecumbens on anti-inflammatory; To study on dose-effect relation and the effectof the active part of Ajuga decumbens on TGF-βexpressions in lung tissues andthe levels of serum interleukin-4 in bleomycin-induced pulmonary fibrosismodel in mice: To investigate acute toxicity of the active part of Ajuga decumbenson mice for offering evidence of clinical safe dose.Method①Water-decocted liquid of Ajuga decumbens was isolated withdistilled water and different concentrations of alcohol by using the way ofmacroporous resin column chromatography respectively. Some researches were madeof the extracted mixture for Ajuga decumbensfrom water-decocted liquid, watereluant and alcohol eluant by using the law of test tube and TLC.②50 mice wererandomly divided into control group, the positive drug group and the experimentalgroup(water-decocted liquid, water eluant and alcohol eluant group). A acuteinflammatory effects of different extracts were observed on the models ofswelling of the earflaps of mice induced by xylene. A chronic inflammatoryeffects of different extracts were observed on the models of placing agar intoabdominal cavity of mice to induce glaucoma. After continuously filled to thestomach of the mice to the dosage of 6g/kg/d, for 3 days of acute inflammatoryexperiment and 14 days of chronic inflammatory experiment respectively. Miceear swelling degree and glaucoma weight was recorded to evaluate theanti-inflammatory effect of the extraction from Ajuga decumbens.③Thesensitivity of alcohol eluant from Ajuga decumbenst against Staphylococcusaureus, Escherichia, Hemolyticstre ptococcus were tested by using punchingand continuously diluting of curette technology. An acute inflammatory wereobserved on the models of swelling of the earflaps of mice induced by xylem. A chronic inflammatory effects were observed on the models of placing agarinto abdominal cavity of mice to induce glaucoma. Then to evaluate thebacteriostasis and anti-inflammatory of alcohol eluant from Ajuga decambens.50 mice were randomly divided into control group, the positive drug group andthe alcohol eluant group. After alcohol extract of Ajuga decumbent wascontinuously filled to the stomach of the mice to the dosage of 3g/kg/d, 6g/kg/dand 12g/kg/d for 3 days of acute inflammatory experiment o and 14 days of chronicinflammatory experiment respectively. Mice ear swelling degree and glaucomaweight was recorded.④72 mice were randomly divided into six groups (12 micein each group). mice in sham-model control group, the model control group,positive medicine group and high, moderate, low dosage of alcohol eluant ofAjuga decumbens groups were injected with bleomycin by trachea. From the nextday, alcohol eluant of Ajuga decumbens solution of differentdosage(3, 6, 12g/kg)was respectively given to the high, moderate and low dosageof Ajuga decumbens alcohol eluant group by ig, while equal volume of normal salinewas given to those in the sham-model control group and model control group andan equal volume of prednisone(30mg/kg)was given to those in positive medicinecontrol group. The mice were killed in turn at 7 and 28 day, lung tissue andblood serum were collected. the right lung was incised to make pathologicalsections which were stained with Hematoxylin—Eosin(HE) for pathologicaldiagnosis, the expression of TGF-βin the lung tissues were analyzed byimmunohistochemistry. Enzyme-linked immunosorbent assay (ELISA) were used todetect the levels of serum IL-4.⑤alcohol eluant of Ajuga decumbent in thehighest concentration and largest amount was given to the mice by stomachfeeding, then behavioral indexes and toxicity degree were continuously observed.In the end of the test, the mice were executed in order to get primary toxicitymaterials.Results①The chemical constituents of Ajuga decumbens was primarilydemonstrated according to precipitation reaction or color reaction with various indicator.②Alcohol eluant of Ajuga decumbent can significantly inhabit miceear swelling and granuloma compared with control(P＜0.01). It also shown to havemore inhibition on acute and chronic inflammation than other fractions.③Alcoholeluant of Ajuga decumbent exerted bacteriostatic action on Staphylococcusaureus,Escherichia and Hemolyticstre ptococcus in vitro, MIC was125mg/ml, 250mg/ml and 500 mg/ml respectively and indicated that both large andmiddle dose of it can significantly inhabit mice ear swelling and granulomacompared with control(P＜0.01).④the high, moderate dosage of Ajuga decumbensalcohol eluant obviously alleviated mice alveoli and fibrosis. TGF-βof themodel group were significantly increased as compared the normal control andthe high, moderate dosage of treatment group (P＜0.01). The levels of serum IL-4of the model group were significantly higher than those of the control groupand the high, moderate dosage of treatment group (P＜0.01), but no effect wasseen in the low dosage of treatment group.⑤all mice healthily survived duringthe period of test and no obvious symptoms were observed. The maximum tolerancedose was as 1440 times as that used in clinic. Observed by naked eyes, therewere no abnormal changes on surfaces and transects of organs.Conclusion①Ajuga decumbens may contain constituents, alkaloids,flavones and glycosides, ploysugars and orgainc acid etc.②Preliminaryclarification of the active part from Ajuga decumbens is alcohol extract.③Both large and middle dosage of alcohol eluant of Ajuga decumbent has distinctlyanti-inflammatory and bacteriostasis therapeutic actions, with thedose-dependence of the effect.④It indicates that the high, moderate treatmentgroup can treat pulmonary fibrosis through reducing the expression of TGF-βinlung tissues and the levels of serum interleukin-4.⑤It indicates no acutetoxicity is found with alcohol eluant of Ajuga decumbent and it is safe to beused in clinic.
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