| Objective: To observe the effect of moxibustion at different times on the levels of uterine tissue microvessel density,prostaglandin F2?(PGF2?),prostaglandin E2(PGE2),vasopressin(AVP),nitric oxide(NO),endothelin(ET-1)and plasma Thromboxane B2(TXB2)and 6-keto-prostaglandin F1?(6-keto-PGF1?)in rat with cold-damp coagulation type dysmenorrhea.To explore its difference in the analgesic effect of the model of primary dysmenorrheal and ite possible mechanism.In order to provide the best time for the clinical choice of moxibustion interventional treatment of cold-damp coagulation type of primary dysmenorrhea and to provide a reliable experimental basis.Methods:In this study,60 Wistar female rats were fed for 7 days by vaginal smear method.45 rats were randomly divided into control group,model group,pre-moxibustion group(Pre-Mox),immediate moxibustion group(Imm-Mox)and pre-immediately moxibustion group(P&I-Mox)(9 rats in eatch group).The rats in the control group were given subcutaneous injection of saline,1times/d,for 10 days(the 1th day and 10 th day injection of 0.5 mg each,2~9 d0.2 mg each),the other 4 groups of cold-damp stagnation type PD rats were treated with(0 ± 1)? ice water bath combined with estradiol benzoate injection method to prepare the model of dyspepsia dysmenorrheal,(0 ± 1)?ice water mixture(room temperature 23 ± 2?)for cold stimulation,1 times/d,20 min/times,while the rats were given subcutaneous injection of estradiol benzoate,1 times/d,for 10 days(the 1th day and 10 th day injection of 0.5 mg each,2~9 d 0.2 mg each).The rats in the control group were given subcutaneous injection of the same dose of saline.Pre-moxibustion groupbegan treatment at the 8th day of the model,the end of treatment on the 10 th day,a total of 3 d.Immediate moxibustion group in the 11 th after the injection of oxytocin(dysmenorrheal model after the success)to start treatment,a total of 1 day.Pre-immediate moxibustion group began treatment at the 8th day of the model,the 11 th day end of treatment,a total of 4 day.Three moxibustion groups were used 7 mm thickness of the moxa,in the Shenque,Guanyuan at the same time temperature and moxibustion,10 min/hole,1 times/d.Control group,model group were used with the same position of the moxibustion group fixed,10 min/times,1 times/d.In addition to the control group,the other 4 groups of cold-damp stagnation type PD rats were injected with oxytocin to observe the behavioral response,to be completed after the behavioral test,the rats were decapitated,quickly separated from the ice tray“Y” shape of the uterus left uterine tissue,washed with cold saline dry wipe dry weighing,adding saline grinding,made of 30% of the uterine tissue homogenate,centrifuge at 4 ? 3000 rpm for 15 min,take the supernatant.The contents of PGE2,AVP and ET-1 in the uterus were measured by radioimmunoassay.The levels of PGF2? in the uterus were measured by enzyme-linked immunosorbent assay(ELISA).The contene of NO in the uterus was measured by biochemical method.The uterine tissue of the right side was fixed with 4% paraformaldehyde solution,paraffinembedded,sliced,part of the uterine tissue MVD was detected by immunohistochemistry,and the remaining part was used for HE staining.Morphological changes of uterine tissue MVD and uterine tissue cells.Results:1 The effect of moxibustion at different times on the behavior of cold-damp coagulation type of PD rats.The results of writhing behavior show: after moxibustion intervention,the latency of pre-moxibustion group,immediate moxibustion group and pre-immediate moxibustion group were significantly prolonged,the number of writhing decreased and the total score of writhing decreased,and the difference was statistically signinficant(P < 0.01).Compared with the other 3moxibustion groups,the number of writhing and the total score of writhing were significantly decreased in the immediate moxibustion group and pre-moxibustion group(P < 0.01);the latency of pre-immediate moxibustion group was significantly longer than that of pre-moxibustion group(P < 0.01).Compared with the immediate moxibustion group,the latency of the rats in the pre-immediate moxibustion group was significantly longer,and the number of writhing and the total score of writhing were significantly reduced(P < 0.05 or P < 0.01).2 The effect of moxibustion at different times on the pathomorphology of uterine with cold-damp coagulation type of PD rats.Uterine tissue HE staining,placed under the lighe microscope,the general morphological observation shows: the control group of rats with morphological rules,no edema,a large number of neutrophils;the model group,the endometrium of the model group was thin,the tissue were seriously edema,congested,a large number of neutrophils and lymphocytes.The pre-moxibustion group,immediate moxibustion group and pre-immediate moxibustion group uterine tissue morphology rules,mild tissue edema,no congestion,a large number of neutrophils,a small amount of lymphocytes.3 The effects of moxibustion at different times on MVD of cold-damp coagulation type of PD rats.MVD of uterine tissue in model group was significantly decreased than that in control group,the difference was statistically significant(P < 0.05).Compared with the model group,the MVD of the uterine tissue of the three moxibustion groups were significantly increased,the difference was statistically significant(P < 0.01).Compared with the pre-moxibustion group,the MVD of the pre-immediate moxibustion group was significantly higher than that of the pre-moxibustion group(P < 0.05).Compared with the pre-moxibustion group,the MVD of the immediate moxibustion group was significantly higher than that of the pre-moxibustion group,but there was no significant difference(P > 0.05).Compared with the immediate moxibustion group,there was no significant difference in the pre-immediate moxibustiongroup,but there was no significant difference(P > 0.05).4 The effects of moxibustion at different times on the levels of TXB2 and6-keto-PGF1? in plasma of cold-damp coagulation type of PD rats.The levels of TXB2 and TXB2/6-keto-PGF1? in the model group were significantly increased,and the content of 6-keto-PGF1? was significantly decreased in the model group(P < 0.01),and the difference was statistically significant(P < 0.01).Compared with the model group,the levels of TXB2 and TXB2/6-keto-PGF1? in the three moxibustion groups were significantly lower than those in the moxibustion group,and the content of 6-keto-PGF1?was significantly increased,the difference was statistically significant(P <0.01).Compared with the pre-moxibustion group,the content of TXB2 in the plasma and the ratio of TXB2/6-keto-PGF1? of the pre-immediate moxibustion group was significantly decreased,the content of 6-keto-PGF1? was significantly increased(P < 0.05 or P < 0.01).Compared with the pre-moxibustion group,the levels of plasma TXB2,6-keto-PGF1? and TXB2/6-keto-PGF1? in the immediate moxibustion group were numerical differences than those in the pre-moxibustion group,but there was no significant difference(P > 0.05).Compared with the immediate moxibustion group,the content of TXB2 and the ratio of TXB2/6-keto-PGF1? in the pre-immediate moxibustion group were significantly decreased(P < 0.01),and the content of 6-keto-PGF1? was significantly higher(P < 0.01).5 The effects of moxibustion at different times on the contents of PGF2?,PGE2,and PGF2?/PGE2 in uterine tissue of cold-damp coagulation type of PD rats.The levels of PGF2? and PGF2?/PGE2 in the uterus were significantly higher than those in the control group,and the contents of PGE2 was significantly decreased in the model group.Compared with the control group,the differences were statistically significant(P < 0.01).Compared with the model group,the levels of PGF2? and PGF2?/PGE2 in the uterine tissue of the three moxibustion groups were significantly lower,the levels of PGE2 was significantly increased high(P < 0.01 or P < 0.05).Compared with thepre-moxibustion group,the levels of PGF2? and PGF2?/PGE2 in the pre-immediate moxibustion group were significantly lower(P < 0.01),and the contents of PGE2 was significantly increased(P < 0.01).Compared with the pre-moxibustion group,the levels of PGF2? and PGF2?/ PGE2 in the immediate moxibustion group were significantly lower(P < 0.05),and the contents of PGE2 was significantly increased(P < 0.01).Compared with the immediate moxibustion group,the levels of PGF2? and PGF2?/PGE2 in the uterus were significantly lower(P < 0.01),and the contents of PGE2 was significantly increased(P < 0.01).6 The effects of moxibustion at different times on the contents of AVP in uterine tissue of cold-damp coagulation type of PD rats.The content of AVP in the uterine tissue of the rats in the model group was significantly higher than that in the control group(P < 0.01).Compared with the model group,the AVP content of uterine tissue in the three moxibustion groups were significantly lower than that in the model group,the difference was statistically significant(P < 0.05 or P < 0.01).Althoug there were numerical differences in the AVP contents between the three groups,but no statistically significant(P > 0.05).7 The effects of moxibustion at different times on the contents of ET-1and NO in uterine tissue of cold-damp coagulation type of PD rats.Compared with the control group,the content of ET-1 in uterine tissue was significantly increased and the content of NO was significantly decreased in the model group,and the difference was statistically significant(P < 0.01).Compared with the model group,the content of ET-1 in the uterus tissue of the three moxibustion groups were significantly lower and the content of NO was significantly increased,the difference was statistically significant(P < 0.01).Compared with the pre-moxibustion group,the content of ET-1 in the uterine tissue of the pre-immediate moxibustion group was significantly lower and the content of NO was significantly increased(P < 0.01).Compared with the pre-moxibustion group,the content of ET-1 in the immediate moxibustion group was significantly lower and the content of NO was significantlyincreased(P < 0.05).Compared with the immediate moxibustion group,the content of ET-1 in the uterine tissue of the rats was significantly lower and the content of NO was significantly increased(P < 0.05).Conclusions:1 Pre-moxibustion,immediate moxibustion and pre-immediate moxibustion at different times can significantly alleviate the cold-damp stagnation type of PD rats abdominal contraction and writhing reaction,significantly prolong the incubation period,but its effect is particularly in front of the most obvious moxibustion,pre-immediate moxibustion.The results showed that the moxibustion had a good analgesic effect on the model of cold-damp stagnation type PD model,but the effect of pre-immediate moxibustion group was more significant.2 Three different time moxibustion can improve the cold-damp stagnation of PD model rats uterine tissue congestion,edema.However,there was no significant difference in histopathological changes between the three groups between pre-moxibustion,immediate moxibustion and pre-immediate moxibustion.3 Three different time moxibustion can increase the cold-damp stagnation type PD model rats uterine tissue MVD,effectively improve uterine local microcirculation.But in particular,in advance to the best effect of moxibustion group,was better than pre-moxibustion and immediate moxibustion.Its role may be through the warmth of the fire,the meridian heat transfer and nerve transmission of the uterus to produce thermal effects to increase blood flow,change the local blood circulation,and thus achieve the purpose of alleviating pain.4 Three different time moxibustion can reduce the plasma TXB2 content and the ratio of TXB2/6-keto-PGF1? in the model of PD model rats,and increase the content of 6-keto-PGF1?.Especially in the most pre-immediate moxibustion.Indicating that the different time moxibustion on the above indicators have a good level of regulation,but the effect of pre-immediate moxibustion regulation is better.Its mechanism may be through the regulationof platelet activity and vascular tension,improve blood vessel contraction,the uterus local blood flow increased,and to achieve the purpose of pain.5 Three different time moxibustion can reduce the levels of PGF2? and PGF2?/PGE2 in the uterine tissue of the model rats of cold-damp stagnation type PD model,and increase the level of PGE2.It was suggested that moxibustion had a good effect on PGF2? and PGE2,and the regulation of moxibustion was better.It was possible to restore the balance of PGF2? and PGE2,improve the endocrine environment and alleviate uterine ischemia and hypoxia state,play a role in analgesia.6 Three different time moxibustion can reduce the AVP content of uterine tissue in the model of cold-damp stagnation type PD model.The mechanism may be to reduce the PG synthesis and release,improve local blood supply,improve myometrial spasmodic contraction state,and thus ease the spastic pain state.7 Three different time moxibustion can adjust the ET-1 and NO of uterine tissue imbalance in the model of cold-damp stagnation type PD model.And the effect of pre-immediate moxibustion is more obvious.The mechanism may be to decrease the content of ET-1 and increase the content of NO,relaxation of vascular smooth muscle,increased uterine blood flow,and then adjust the uterine contraction,relieve pain. |
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