The Study Of Antifertility And Anti-inflammatory Effects Of Compound Kucen Gel

Part Ⅰ Study on the spermicidal effect of alcohol extracts with different ratios of Sophora flauescens Ait and Chinese Bulbul in vitroObjective:To observe the spermicidal effect and mechanism of alcohol extracts with different ratios of Sophora flauescens Ait and Chinese Bulbul in vitro.Methods:1. Sperm aseptically obtained by masturbation and prepared by density gradient centrifugation from15healthy men were incubated in alcohol extracts with nine different ratios of Sophora flauescens Ait and Chinese Bulbul respectively. Measurements of sperm motility and motility parameters were carried out at20s,2min and4min in all specimens by computer-assisted sperm analysis (CASA).2. Minimal effective concentrations of the spermicidal effect of alcohol extracts with the nine ratios were also studied using the above semen samples at20s.3. Transmission electronic microscope was used to observe the morphological changes of sperms incubated by alcohol extracts with the best ratio of Sophora flauescens Ait and Chinese Bulbul.Results:1. At ratio3:1of Sophora flauescens Ait and Chinese Bulbul, the alcohol extract can significantly inhibit the sperm motility (grade A and B) at the concentration of0.5mg/mL. And the inhibition is more obvious with increasing time. Meanwhile, at ratio3:1, it also can significantly inhibit human sperm motility parameters including curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity (VAP), amplitude of lateral head displacement (ALH), wobble (WOB) and mean angular displacement (MAD), compared with the control group.2. At ratio3:1, the alcohol extract has the lowest minimal effective concentration of the spermicidal effect with the concentration of3.5mg/mL.3. After the incubation with alcohol extracts at3:1ratio of Sophora flauescens Ait and Chinese Bulbul, sperm cell membranes and sperm acrosome of the semen samples were destroyed through the observation of transmission electronic microscope.Conclusions:At ratio3:1of Sophora flauescens Ait and Chinese Bulbul, the alcohol extracts have the best spermicidal effect in vitro. And the mechanism may be related to the destruction of the sperm cell membranes and sperm acrosome. Part Ⅱ In vivo study of antifertility effect of compound Kucen gelObjective:To study the antifertility effect and vaginal irritation test of compound Kucen gel in vivo.Methods:1. Sixty female Sprague Dawley (SD) rats of8weeks old were randomly divided into six groups:saline group, blank gel group, compound Kucen gel low (0.05g crude drug/g), medium (0.10g crude drug/g), high dosage (0.15g crude drug/g) groups as well as nonoxinol group (positive control group). Then these drugs (200μL) were separately injected intravaginally to nonpregnant rats before copulation with male rats at the ratio1:1. After successfully mating with male rats, the female rats were dissected to calculate embryos and contraception rates on day12.2. Fifteen female SD rats of8weeks old were randomly divided into three groups (saline group, blank gel group and compound Kucen gel high dosage group). The above drugs (200μL) were injected intravaginally to the rats respectively, once every day for two weeks. After the last administration, rats were sacrificed to examine the vagina, and also do the further pathological examinations.Results:1. The compound Kucen gel high dosage group (0.15g crude drug/g) can significantly inhibit the number of embryos of female rats when compared with the saline group,(0.00±0.00vs11.00±2.00, P<0.05), and statistically insignificant with the positive control group (0.00±0.00vs0.00±0.00, P>0.05). The contraception rate of high gel dosage group is100%.2. There were neither vagina congestion nor swelling found in all threes groups after2weeks’ administration. The further pathological examinations demonstrated that the vaginal tissues were normal without inflammatory cell infiltration. Conclusions:Compound Kucen gel had strong antifertility effect and no toxic effect was found by the vaginal toxicity test. Part ⅢIn vivo study of anti-inflammatory effect of compound Kucen gelObjective:To study the anti-inflammatory effect of compound Kucen gel in vivo.Methods:The anti-inflammatory activity was evaluated by animal model using xylene-induced ear edema in mice. Sixty Kunming mice were randomly divided into6groups:saline group, blank gel group, compound Kucen gel low (0.05g crude drug/g), medium (0.10g crude drug/g), high dosage groups (0.15g crude drug/g) as well as positive control group (clobetasol propionate cream group). The above drugs (50μL) were applied to the posterior and anterior surfaces of the right ear of mice, respectively, once every day for four consecutive days. The left ear was considerd as a control. After the last administration,50μL of xylene was also applied to the posterior and anterior surfaces of the right ear of each mouse. One hour after xylene application, the mice were sacrificed to access the ear swelling degree and the average inhibition rate.Results:The high dosage group of compound Kucen gel can significantly inhibit the swelling degree when compared with the saline group (10.17±2.56vs21.32±3.17, P<0.01), and statistically insignificant with the positive control group (10.17±2.56vs8.53±1.89, P>0.05). And the inhibit rate is52.3%.Conclusions:Compound Kucen gel has strong anti-inflammatory effect.