| Background:Primary dysmenorrhea(PD),also known as functional dysmenorrhea,refers to menstrual pain with insignificant pathological findings of reproductive organs found through gynaecological examination.While the incidence of PD is increasing gradually,it has negative impacts on women’s physical and mental health.The pathogenesis of PD is relatively complicated and closely associated with prostaglandins,vasopressin,oxytocin,β-endorphin,estradiol,progesterone,calcium ion,magnesium ion,nerve and neurotransmitter,endothelin and nitric oxide.At present,the more commonly accepted theory of pathogenesis involves changes in contents of prostaglandin E2 and prostaglandin F2a.The common treatment options in western medicine include oral non-steroid anti-inflammatory medicines and contraceptives,which are known to have multiple side-effects and found to have insignificant effect in the long run.While treatment options such as oral calcium channel blocker,vitamins,transcutaneous electric nerve stimulation,psychological therapy and aerobic exercise are proven effective in the short-term,their long-term effects remain limited.Surgical intervention though uncommon,is usually recommended for recurrent PD patients who are unresponsive to oral medications,PD fits into the category of "dysmenorrhea during menstruation" in Traditional Chinese Medicine(TCM),and its pathogenesis in TCM can be summarized as "blockage causes pain" and"unnourishment causes pain".While TCM treatment of PD is known to be curative,different treatment options such as herbal medications,acupuncture,massage,point-embedding therapy,point-application etc have varying effects.Abdominal vibration tuina,which is a form of massage mainly performed on the abdomen,uses "song-zhen manipulation" as its basic technique to give rise to abdominal vibrational form of zang-fu massage therapy.It is known to help regulate Yin and Yang,tonify to Innate qi,warm Yang and disperse cold etc and its effect on PD has known to be curative.However,the underlying mechanism for its effect on PD is yet to be elucidated.Thus we aim to do a thorough research on its underlying mechanism through this project.Purpose:This research by establishment of PD murine model,used enzyme-linked immunosorbent assay and immunohistochemical methods etc to evaluate responses to abdominal vibration tuina between model group and drug group murine models.We then analyzed the treatment’s effects on the levels of PGF2a and PGE2 and the activity change of PGF2a receptors in the PD model.Lastly,we investigated the underlying mechanisms of abdominal vibration tuina in treatment of PD based on the levels of PGE2,PGF2a.Methods:Forty healthy female SD mice at the age of 8 weeks underwent a week of adaptive feeding before a vaginal smear was done to confirm status of 10-day female estrous cycle via cell morphology,species and quantity.The mice with confirmed status of 10-day female estrous cycle continued to have vaginal smear in order to choose anestric female mice.32 mice were eventually chosen and evenly randomized into blank,model,positive drug and tuina groups;with 8 mice in each group.PD murine models were prepared using a combination of estradiol benzoate and oxytocin.In the model,positive drug and tuina groups,the estradiol was injected into the abdomen of each mouse for 10 consecutive days.The dose was 0.5mg/d in the first and tenth days,and 0.2 mg/d in the second to ninth days.On the 10th day,intraperitoneal injection of oxytocin at 2U was given to all mice.Torsional reaction was observed and recorded.After the establishment of PD murine models,mice from positive drug group were given celecoxib at 30mg/1000g/d by gavage.While mice from tuina group were treated with abdominal vibration tuina for 15min per day for 10 consecutive days.Observation indexes:general behavior and writhing response of mice;levels of PGE2,PGF2a in serum and uterine tissue;PGF2a receptor(PTGFR)level.Results:1.Comparing with mice in other groups,PD mice in model group showed a dimmer shade of withered-yellow hair,slower in action and behavior,lower in metabolism and weaker mental status.These symptoms showed improvement after treatment in mice of positive drug and tuina group.2.Comparing with mice in blank group,the latent time of torsion body for the mice in the other three groups is shorter,and the number of torsion is increased(P<0.01),indicating that the successful establishment of PD mouse models.The latency of the torsional body of mice in positive drug group and tuina groups are significantly longer than that of the model group,and the number of torsion are significantly reduced(P<0.05).Comparing with mice in tuina group,the latency of torsion body of mice in positive drug group may have increased and the number of torsion decreased,but the differences between the two groups was found to be statistically insignificant.3.Comparing with mice in blank group,the levels of PGF2a and PGE2 in serum of mice in model group were proven to be statistic-significantly different(P<0.05).Comparing with mice in model group,the serum PGF2a levels in mice from positive drug and tuina groups decreased,and the serum PGE2 levels of tuina group increased;these differences were proven to be statistically significant(P<0.05).4.Comparing with mice in blank group,the levels of PGF2a in the endometrium of mice from the other three groups showed an increase,while the levels of PGE2 in the endometrium of the mice from model and positive drug groups showed a decrease in comparision;differences were found to be statistically significant(P<0.05).Comparing with mice from model group,the levels of PGF2a in mice from positive drug and tuina groups showed a decrease,while the PGE2 level in mice from tuina group showed an increase;with differences proven to be statistically significant(P<0.05).Furthermore,there is a statistically significant(P<0.05)increase in levels of PGE2 in mice from tuina group than that in positive drug group.5.Comparing with blank group,the expression of PTGFR in model group showed more significant increase(P<0.05).Comparing model group with positive drug group and the tuina group,mice in the later groups had statistically-significant decrease in the expression of PTGFR(P<0.05).Conclusion:1.Abdominal vibration tuina has obvious improvements on the general condition of PD mice models such as a prolonged latent time of torsion body,reduction in the number of times of torsion,demonstrating an improvement on the symptoms of dysmenorrhea.2.Abdominal vibration tuina can reduce the levels of PGF2a in serum and endometrium,while increasing the levels of PGE2,thus inhibiting the contraction of smooth muscle of uterus,promoting the relaxation of uterine smooth muscles and giving relief to dysmenorrhea symptoms.3.Abdominal vibration tuina plays a role in the treatment of dysmenorrhea by inhibiting the expression of PTGFR in the endometrium reduces its combination with PGF2a,thus reducing spastic contraction of uterine smooth muscle.In summary,we found the effects of abdominal vibration tuina therapy on PD to be significant and its underlying mechanism involves a decrease in levels of PGF2a in serum and endometrium,increase in levels of PGE2,inhibition of PTGFR expression in the endometrium,thus reducing uterine smooth muscle contraction,contributing to its treatment for PD. |
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