| Partâ… Effects of Bushen Treatment and Shugan Treatment on Ovulation Number in Gonadotropin Pretreatment MiceObjective: To study the effects of BushenTiaojingâ…¡Recipe,â…¢Recipe and Xiaoyao Pill on ovulation number of Gonadotropin pretreatment mice.Methods: One hundred and two female immature mice were randomly divided into 17 groups with 6 mice each group. 1 to 4 were Bushen Treatment groups, 5 to 8 were Shugan Treatment groups, 9 to 12 were BushenTiaojingâ…¡Recipe add Xiaoyao Pill groups, 13 to 16 were Xiaoyao Pill add BushenTiaojingâ…¢Recipe groups, 17 was control group. All groups except control group were treated by BushenTiaojingâ…¡Recipe (high or low does) or Xiaoyao Pill (high or low does) 0.4 ml or during the 1-3 day to promot follicular development. All groups were injected PMSG (5IU/each) by intraperitoneal at 1pm 3rd day, and then with HCG (5 IU/each) after 48 h. All groups except control group were treated with 0.4 ml BushenTiaojingâ…¢Recipe (high or low does) or Xiaoyao Pill (high or low dose) on the 4-5 day. Mice were executed at 8am of the 6th day, and ampulla portion of uterine tubes were punctured with somatotype microscope to count the number of ovulated oocytes.Results: Compared with control group by One way ANOVA, ovulation number was more in BushenTiaojingâ…¡Recipe high-dose add BushenTiaojingâ…¢Recipe high-dose of Bushen Treatment(P=0.003) and Xiaoyao Pill high-dose group add Xiaoyao Pill low-dose group of Shugan Treatment(P=0.007). Factorial experiment design analysis in each group, treatment BushenTiaojingâ…¡Recipe high-dose and low-dose group had significant difference(P=0.004), and treatment Xiaoyao Pill high-dose group and low-dose group in induced ovulation had significant differenc(eP=0.039), and they have interaction(P=0.047, P=0.046). Compared with control group by one way ANOVA, ovulation number in Xiaoyao Pill low-does addâ…¢Recipe high-does group andâ…¡Recipe high-does add Xiaoyao Pill low-does group were significantly more(P=0.013, P=0.039), but they had no interaction by factorial design analysis(P=0.838, P=0.604).Conclusions: Ovulation number in BushenTiaojingâ…¡addâ…¢Recipe high-dose group was more, related to YinJing was magnificent to promote follicle's development and YangQi was easy to transformation between Yin and Yang. Xiaoyao Pill high-dose promoted follicular development, while low-dose induced ovulation, and ovulation number in this group was the most. Jing and Xue was magnificent to ovarian follicle's development; adjusted liver was useful to transformation between Yin and Yang. Two methods had interaction, so can improve curative effect.Partâ…¡Effects of Bushen Treatment and Shugan Treatment on COX-2,PTX3,TSG-6 in Gonadotropin Pretreatment Mice OvaryObjective: To study and compare the effects of Bushen Treatment and Shugan Treatment on COX-2,PTX3,TSG-6 of gonadotropin pretreatment mice ovary in ovulatory processes.Methods: Ninety-six female immature KM mice were randomly divided into normal group, control group, Bushen group and Shugan group of 4 mice. Bushen group and Shugan group were treated with BushenTiaojingâ…¡Recipe high-dose and Xiaoyao Pill high-dose respectively for 3 days from experiment started, while normal group and control group gave distilled water. All groups except normal group were injected intraperitoneally with PMSG (5 IU/each) at 8am on the 3rd day, and 48h later injected with hCG (5 IU), and the normal group injected with distilled water at the same time. On the 4-5 day, Bushen group and Shugan group were treated with BushenTiaojingâ…¢Recipe and Xiaoyao Pill low-does 0.4 ml respectively, and normal group and control group were treated with distilled water 0.4 ml. Mice were executed respectively at 0h,4h,8h,12h after injecting hCG, and ovaries were preserved in liquid nitrogen. Examined expression of COX-2,PTX-3 mRNA with RT-PCR and COX-2,TSG-6 protein with Immunohistochemical method.Results:1 Expression of COX-2 mRNA and protein in each group at 0h, 4h, 8h, 12 h after injecting hCG Expression of COX-2 was increased at 0h,4h,8h after injecting hCG,and decreased at 12h. Compared with control group, expression of COX-2 in Bushen group and Shugan group were significantly increased(P<0.05). Expression of COX-2 in Bushen group was decreased but had no significant difference(P>0.05)at 0h after injecting hCG. At 4 h, expression of COX-2 in Bushen group and Shugan group were significantly decreased(P<0.05)when compared with control group. Expression of COX-2 in Bushen group was decreased but had no significant difference(P>0.05)when compared with Shugan group. At 8h, compared with control group, expression of COX-2 in Shugan group were significantly increased(P<0.01), and Bushen group was decreased(P<0.05). Compared with Shugan group, expression of COX-2 in Bushen group was significantly decreased(P<0.01). At 12h, compared with control group, expression of COX-2 in Shugan group were significantly increased(P<0.05), Bushen group was decreased but had no significant difference(P>0.05). Compared with Shugan group, expression of COX-2 in Bushen group was significantly decreased(P<0.05). 2 Expression of TSG-6 protein expression in each group at 0h, 4h, 8h, 12h after injecting HCG Expression of TSG-6 protein in control group was gradually increased, but had no significant difference between 4h and 8h(P>0.05). Expression TSG-6 protein in Shugan group was gradually increased, but had no significant difference between 8h and 12h(P>0.05). At 0h, expression TSG-6 protein in Bushen group was gradually increased, and all group had significant difference(P<0.01). Expression TSG-6 protein in Bushen group and Shugan group had no significant difference(P>0.05)when compared with control group. At 4h, expression TSG-6 protein in Bushen group and Shugan group had no significant difference(P>0.05)when compared with control group, and in Bushen group was significantly increased(P<0.05)when compared with Shugan group. At 8h, expression TSG-6 protein in Bushen group and Shugan group were significantly increased(P<0.01)when compared with control group, and in Bushen group was decreased but had no significant differenc(eP>0.05)when compared with Shugan group. At 12h, expression TSG-6 protein in Bushen group and Shugan group were significantly increased(P<0.05, P<0.01)when compared with control group, and expression TSG-6 protein in Bushen group was significantly increased(P<0.01)when compared with Shugan group.3 Expression of PTX-3 mRNA in each group at 0h, 4h, 8h, 12h after injecting hCG Expression of PTX-3 mRNA in control group was gradually increased, and had significant difference among 0h,4h,8h after injecting hCG(P<0.01). Expression of PTX-3 mRNA at 12 h was significantly decreased(P<0.01)when compared with that at 8h. At 0h,4h, expression of PTX-3 mRNA in Bushen group and Shugan group were significantly increased(P<0.01 ) when compared with control group, and in Bushen group was significantly decreased(P<0.01)when compared with Shugan group. At 8h, expression of PTX-3 mRNA in Bushen group and Shugan group were significantly decreased(P<0.01)when compared with control group, and in Bushen group was significantly decreased(P<0.01)when compared with Shugan group. At 12h, expression of PTX-3 mRNA in Bushen group and Shugan group were significantly increased(P<0.01)when compared with control group, and in Bushen group had no significant difference(P>0.05)when compared with Shugan group.Conclusions: Bushen Treatment can up-regulate COX-2 expression, while Shugan Treatment reduce it. Both of Bushen Treatment and Shugan Treatment could up-regulate expression of TSG-6 protein and PTX3 mRNA. Both of Bushen Treatment and Shugan Treatment could up-regulate expression of TSG-6 protein, and this maybe one reason to influence ovulation number. Partâ…¢Effects of Bushen Treatment and Shugan Treatment on ADAMTS-1,Cat L in Gonadotropin pretreatment mice OvaryObjective: To study the effects of Bushen Treatment and Shugan Treatment on the expression of ADAMTS-1,Cat L mRNA and protein of Gonadotropin pretreatment mice. To analyze the mechanisms of those two treatments in prompting follicle break.Methods: Group and materials are the same as that in partâ…¡. Reverse transcription-polymerase chain reaction (RT-PCR) was used to semiquantitatively detect the expression of ADAMTS-1 mRNA and Cat L mRNA during ovulation, and Western blot was used to detect the expression of the protein. Immunohistochemical method was used to detect the expression of PR, ADAMTS-1 and Cat L protein.Results:1 Expression of PR protein in each group ovary at 0h, 4h, 8h, 12h after injecting hCG At 0h, 8h and 12h after injecting hCG, expression of PR in Bushen group and Shugan group ovary were significantly increased(P<0.01)when compared with control group. At 4h, expression PR protein in Bushen group and Shugan group had no significant difference(P>0.05)when compared with control group. At 0h, 4h, 8h, 12h after injecting hCG, expression of PR protein in Bushen group had no significant difference(P>0.05)when compared with Shugan group. 2 Expression of ADAMTS-1 mRNA and its protein in each group ovary at 0h, 4h, 8h, 12h after injecting hCG Expression of ovary ADAMTS-1 at these four timings was gradually increased, and was the highest at 12h. Expression of ADAMTS-1 in each group ovary was low at 0h. At 4h, expression of ADAMTS-1 in Bushen group and Shugan group was significantly increased(P<0.05 or P<0.01)when compared with control group. Expression of ADAMTS-1 in Bushen group was decreased, but had no significant difference(P>0.05)when compared with Shugan group. At 8h, compared with control group, expression of ADAMTS-1 in Shugan group was significantly increased(P<0.01), and in Bushen group had no significant difference ( P > 0.05 ) . Compared with Shugan group, expression of ADAMTS-1 in Bushen group was significantly decreased(P<0.01). At 12h, compared with control group, expression of ADAMTS-1 in Shugan group had no significant difference(P>0.05), and in Bushen group was significantly increased(P<0.01). Expression of ADAMTS-1 in Bushen group was significantly increased(P<0.01)when compared with Shugan group. 3 Expression of Cat L mRNA and protein in each group ovary a at 0h, 4h, 8h, 12h after injecting hCG Expression of Cat L in each group ovary at these four timings was gradually increased, and was the highest at 12h. Expression of Cat L in each group ovary was low. At 4h, expression of Cat L in Bushen group and Shugan group was significantly increased(P<0.01)when compared with control group. Expression of Cat L in Bushen group was decreased, had no significant difference(P>0.05)when compared with Shugan group,. At 8h, expression of Cat L in Shugan group was significantly increased(P<0.01)when compared with control group, and in Bushen group had no significant difference(P>0.05). Compared with Shugan group, expression of Cat L in Bushen group was significantly decreased(P<0.01). At 12h, compared with control group, expression of Cat L in Shugan group was no significant difference(P>0.05), and in Bushen group was significantly increased(P<0.01). Expression of Cat L in Bushen group was significantly increased(P<0.01) when compared with Shugan group.Conclusions: Ovary PR protein expression at preovulation of Gonadotropin pretreatment mice was increased with Bushen Treatment and Shugan Treatment. But expression of PR protein at 12h was higher in Bushen group than in Shugan group. Expression of ADAMTS-1 and Cat L of Gonadotropin pretreatment mice ovary was increased with Bushen Treatment and Shugan Treatment. Expression of ovary ADAMTS-1,Cat L was highest at ovulation (hCG injecting 12h )with Bushen Treatment, while expression was quickly increased at preovulation (hCG injecting 8h) then slow-moving to the highest at ovulation with Shugan Treatment. Peak of ADAMTS-1,Cat L expression in Bushen group was higher than in Shugan group. |
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