The Experimental Research For Angelica-Chuanxiong In The Treatment Of Primary Dysmenorrheal Model

Objective: As one of the most common gynecology diseases, Primary Dysmenorrhea?PD? is clinically typical of periodical lower abdomen pains and heavy periods, which are the main symptoms. Females with severe dysmenorrhea have worse life quality than those without dymenorrhea. As we've known, protaglandin?PG? is tightly bonded with gynecology pains. In recent years, researches reveal that PGF₂? content variation is very crucial to the pathogenesis of PD. Meanwhile, OT, OTR, E2, P, AVP, etc. are factors that can control PGF₂? content variation. This project is supposed to research on the pathogenesis of PD by analyzing expressions of PGF₂?, PGE2, OT, OTR, E2, P, AVP in uterine tissues based on experimental animal models. Also, the experimental mouse would be cured with Angelica-Chuanxiong prescription so as to find out the best compatibility of medicines to prevent the content of PGF₂? increasing as well as the influential mechanism of Angelica-Chuanxiong on PD.Method: 70 selected female Kunming mouse at SPF level, weighing 18-22 g, are divided into different groups:namely the drug 1:1 group?3.03g/kg?,the drug 2:1 group?2.28g/kg?, the drug 3:1 group?2.02g/kg?,the drug 3:2 group?2.58g/kg?,the yimucaogao group?0.60g/kg?, the model group and the blank group. The primary dysmenorrhea model established with progynova?0.5mg/kg? and oxytocin?2IU/mouse? had been dosed for 12 days. The dosing plan was as follows: For the Medicine Group, the matched medicine was gavaged as 20ml·kg-1·d-1 in the morning, while progynova was gavaged in the afternoon; For the medicinal extract of yimucaogao group, the medicinal extract of yimucaogao was gavaged as 20ml·kg-1·d-1 in the morning, while progynova was gavaged in the afternoon; For the model group, the normal saline was gavaged as 20ml·kg-1·d-1 in the morning, while progynova was gavaged in the afternoon; For the blank group, the normal saline was gavaged as 20ml·kg-1·d-1 twice the day. One hour after the progynova was gavaged on the twelfth day, oxytocin, 2IU/per mice?0.2ml?, was injected into the enterocoelia to cause the Writhing response. Meanwhile, only normal saline was injected for the blank group at the level of 0.2ml/per mice. Writhing incubation and writhing frequency were observed 30 minutes after the injection of oxytocin. The animals were killed after the success of behavior indexes observation. Uteruses were peeled, as one part was transferred into the refrigerator at-80? once exposed into liquid nitrogen for the Real Time PCR and Western-blot detection. And the supernate of homogenate from the other part was used for ELISA detection.Result: 1 Writhing response Writhing response happened in most experimental groups except for the blank group, reaching the incidence rate of 100%.Comparing with the model group,the writhing incubation of the drug 1:1 group,drug 2:1 group, drug 3:1 group, drug 3:2 group and yimucaogao group were apparently extended,the difference is good for statistics?P < 0.01?; The writhing frequency were apparently cut down, which is also good for statistics?The drug 2:1 group: P < 0.01, the others: P < 0.05?; the difference among the treatment groups barely has anything good for statistics?P > 0.05?. 2 The protein expression of OTR 2.1 OTR Protein by Western-blot Comparing with the blank group,the expression of OTR protein was increased in the model group and five treatment groups?the model group: P < 0.01,the others: P < 0.05?.Comparing with the model group, the expression of OTR protein was decreased in five treatment groups,the difference is good for statistics?P < 0.01?.The difference among the treatment groups barely has anything good for statistics?P > 0.05?. 2.2 OTR m RNA by Real time PCR Compared with the blank group,the expression of OTR m RNA was increased in the model group and five treatment groups?the model group and the yimucaogao group: P < 0.01,the others: P < 0.05?.Compared with the model group, the expression of OTR m RNA was decreased in five treatment groups?the yimucaogao group: P < 0.05,the others: P < 0.01?.Compared with the yimucaogao group,the expression of OTR m RNA was decreased in the drug 2:1 group?P < 0.05?. The difference among the four medicine groups barely has anything good for statistics?P > 0.05?. 3 Expression of PGF₂?, PGE2, OT, E2, P, AVP by ELISA PGF₂?: Comparing with the blank group,the expression of PGF₂?wasincreased in the model group and five treatment groups?P < 0.01?.Comparing with the model group, the expression of PGF₂? was decreased in five treatment groups?P < 0.01?.Comparing with the yimucaogao group,the expression of PGF₂?was decreased in the drug 2:1 group?P < 0.05?.The difference among the four medicine groups barely has anything good for statistics?P > 0.05?. PGE2: Comparing with the blank group,the expression of PGE2 was decreased in the model group and five treatment groups?The drug 2:1 group: P < 0.05,the others: P < 0.01?.Comparing with the model group, the expression of PGE2 was increased in five treatment groups?P < 0.01?.Comparing with the yimucaogao group,the expression of PGE2 was increased in the drug 2:1 group?P < 0.05?.The difference among the four medicine groups barely has anything good for statistics?P > 0.05?. OT: Comparing with the blank group,the expression of OT was increased in the model group and five treatment groups?The drug 1:1 group,the drug 2:1 group and the drug 3:1 group: P < 0.05,the others : P <0.01?.Comparing with the model group, the expression of OT was decreased in five treatment groups?The yimucaogao group: P < 0.05, the others: P < 0.01?.The difference among the five treatment groups barely has anything good for statistics?P > 0.05?. E2: Comparing with the blank group,the expression of E2 was increased in the model group and five treatment groups?The drug 3:1 group,the drug 3:2 group and the model group: P < 0.01,the others: P < 0.05?.Comparing with the model group, the expression of E2 wasdecreased in five treatment groups?The drug 3:1 group and the drug 3:2 group: P < 0.05, the others: P < 0.01?.While there was no statistical significance among five treatment groups?P > 0.05?. P: Comparing with the blank group,the expression of P was decreased in the model group and five treatment groups?The drug 1:1 group and the drug 2:1 group: P < 0.05,the others: P < 0.01?.Comparing with the model group, the expression of P was increased in five treatment groups?The drug 1:1 group and the drug 2:1 group: P < 0.01,the others: P < 0.05?.The difference among the treatment groups barely has anything good for statistics?P > 0.05?. AVP: Comparing with the blank group,the expression of AVP was increased in the model group and five treatment groups?The drug 2:1 group: P < 0.05, the others: P < 0.01?.Comparing with the model group, the expression of AVP was decreased in five treatment groups?P < 0.01?.Comparing with the yimucaogao group,the expression of AVP was decreased in the drug 2:1 group?P < 0.05?.The difference among the four medicine groups barely has anything good for statistics?P > 0.05?.Conclusion: 1 the model of Primary Dysmenorrhea is successful confirmed by behavior, can be used in the study of clinical disease. 2 PGF₂?,OT,E2,AVP,OTR protein,OTR m RNA are overexpression in the uterus tissue of mouse primary dysmenorrhea model,but PGE2?P are lower expression in the uterus tissue of mouse primary dysmenorrhea model.It shows that PGF₂?, PGE2,OT,E2,P,AVP,OTRprotein and OTR m RNA are all involved in the primary dysmenorrhea-associated pain pathogenesis. 3 Angelica-Chuanxiong can function as abirritation. Furthermore, Angelica-Chuanxiong is able to decrease the the high level of PGF₂?, OT, E2, AVP, OTR Protein and OTR m RNA inside the uteruses of PD model,also increase the low level of PGE2 and P, which suggest that Angelica-Chuanxiong can adjust the level of PGF₂?, PGE2,OT, E2, P, AVP, OTR Protein and OTR m RNA to release primary dysmenorrhea-associated pain. The effect is as well as Yimucaogao.