Changes Of Gynecology Cold Syndrome Model Rats T-PA/PAI-1 And Modified Decoction For Warming Meridian Intervention Effect

Objective:Observe the changes of gynecological cold syndrome model rats serum and ovarian tissue in t-PA,PAI-1 and its relationship with reproductive endocrine; Study on the relationship between the occurrence of gynecological cold syndrome and fibrinolytic fuction; Using Warming meridian decoction intervention, to investigate the effect of the Modified decoction for warming meridian on reproductive function of gynecological cold syndrome model rats.Methods:Healthy clean grade female SD 60 rats, February age, weight 200±20g, adaptive feeding for 1 weeks, female SD rats were randomly divided into normal group, model group and treatment group with 20 rats in each group. Rat model of gynecology excess-cold syndrome was made by ice water immersion. From the first day, those rats of the treatment group were intragastric administration in dose of 3.51g/200g/day seperatively, qd for 2 weeks. The other two groups were given the same voluminal distilled water. After 2 weeks, weighed and sacrificed the rats by decapitation during anestrum. Get blood by decapitation, centrifugalization, separating serum,placed them respectively. dissected the level of E2, P, T, FSH and LH by means of radio immunoassay. dissected the plasma t-PA, PAI-1 level by the means of ABC-ELISA. Dissected and got rats ovary quickly. Fixed right ovary in 4% Polyoxymethylene, dehydrated with alcohol, transparented with dimethyl benzene, embedding, slice in 5 μm, HE dyeing, observed the construre of ovarian histomorpholory under light microscope. The other ovary were placed in labeled plastic freezer tube, dissected the expression of t-PA, PAI-1 protein by the means of Western Blood, dissected the t-PAm RNA、PAI-1m RNA by the means of RT-PCR.Results:1 General state of health: After being frozen, the model rats appeared such symptoms as chilling, im in clothing hari, preference for herd, twisting and less-movement, slow reaction, with dark purple in claws and tail and rotten stool. In treatment group, the above-mentioned symptoms improved to some different degree.2 Smearing of vagina cells:The normal group: The duration of estrous cycle is 100±6.64 h and the diestrus 31.2±11.2 h. With clear regularity and periodicity change.The model group: The duration of estrous cycle is 115±8.29 h and the diestrus 50.4 ± 8.64 h. Compared with normal rats, the estrous cycle lengthened in model group, there were significance difference(P<0.01), he diestrus lengthened, there were significance difference(P<0.01)The treatment group: The duration of estrous cycle is 107±8.18 h and the diestrus 39.6±8.16 h. Compared with model rats, the time of estrous cycle shorter in model group, there were significance difference(P<0.01), and the time of diestrus shorter also, there were significance difference(P<0.01)3 Change of contents of serum E2, P, T, FSH, and LH: Compared with normal group, in model group,the serum E2, P, T, FSH, and LH levels decreased, differences were significant(P < 0.01). Compared with model group, in treatment group, the serum E2, P, T, FSH, and LH levels rose, differences were significant(P < 0.05). Compared with normal group, in treatment group, the serum E2, P, T, FSH, and LH levels decreased slightly, differences were not significant(P>0.05).4 Histomorphology changes in ovary: the appearance of ovary is normal, grades of follicles can be seen on it in normal group. While in model group, there can be seen less grades of follicles and the mature follicle rare. In treatment group, the above-mentioned symptoms improved to some different degree.5 Determination of plasma t-PA, PAI-1 contentCompared with normal group, in model group, the plasma t-PA content decreased, differences were significant(P<0.01). Compared with model group, in treatment group, the plasma t-PA content rose, differences were significant(P<0.05). Compared with normal group, in treatment group, the plasma t-PA content decreased slightly, differences were significant(P<0.01). Compared with normal group, in model group, the plasma PAI-1 content rose, differences were significant(P<0.01). Compared with model group, in treatment group, the plasma PAI-1 content decreased, differences were significant(P<0.05). Compared with normal group, in treatment group, the plasma PAI-1 content rose slightly, differences were significant(P<0.05).6 Expression of t-PA、PAI-1 proteins in ovarian tissueCompared with normal group, in model group, the expression of t-PA proteins in ovarian tissue decreased, differences were significant(P<0.01). Compared with model group, in treatment group, the expression of t-PA proteins in ovarian tissue rose, differences were significant(P < 0.01). Compared with normal group, in treatment group, the expression of t-PA proteins in ovarian tissue decreased, differences were not significant(P>0.05).Compared with normal group, in model group, the expression of PAI-1 proteins in ovarian tissue rose, differences were significant(P < 0.01). Compared with model group, in treatment group, the expression of t-PA proteins decreased, differences were significant(P<0.01). Compared with normal group, in treatment group, the expression of t-PA proteins rose, differences were significant(P<0.05).7 Changes of t-PA m RNA、PAI-1 m RNA in ovarian tissueCompared with normal group, in model group, the expression of t-PA m RNA in ovarian tissue decreased, differences were significant(P<0.01). Compared with model group, in treatment group, the expression of t-PA m RNA in ovarian tissue rose, differences were significant(P < 0.01).Compared with normal group, in treatment group, the expression of t-PA m RNA in ovarian tissue decreased, differences were significant(P<0.01).Compared with normal group, in model group, the expression of PAI-1 m RNA in ovarian tissue rose, differences were significant(P < 0.01). Compared with model group, in treatment group, the expression of PAI-1 m RNA in ovarian tissue decreased, differences were significant(P<0.01). Compared with normal group,in treatment group, the expression of PAI-1 m RNA in ovarian tissue rose, differences were significant(P<0.01).Conclusions:1 Genecological cold syndrome model rats have the reproductive endocrine disorders and ovarian dysfunction.2 Gynecology cold syndrome model rats appeared fibrinoly dysfunction in the whole body and local ovarian.3 Warming meridian decoction can improve the fiber of rat model of cold syndrome, recover the normal reproductive function of ovary and maintain the reproductive endocrine homeostasis.