Based On “Accumulation Due To Asthenic Disease” To Explore The Mechanism Of Yiqi Xiaozheng Recipe In Rats With Uterine Leiomyoma

Objective:To observe the effect of Yiqi Xiaozheng recipe on uterine pathology,T lymphocyte subsets,hemorheology,apoptosis and cycle distribution of myoma cells,ER,PR and Bcl-2,Bax,Caspase-3 protein in uterine tissue of rats with uterine fibroids,and to explore the mechanism of Yiqi Xiaozheng recipe in the treatment of uterine fibroids,so as to provide new ideas and theoretical basis for clinical treatment of uterine fibroids.Method:1.Fifty female SPF Wistar rats were randomly divided into blank group(K,n=8)and model group(n=42)after one week of adaptive feeding.The model group was intraperitoneally injected with estradiol benzoate injection 0.5mg/kg,once a day+progesterone injection 4mg/ kg,once a week.After 25 days,the two hormones were used for 5 days,and the dose remained unchanged;Rats in the blank group were intraperitoneally injected with the same volume of normal saline.After successful modeling,the rats were randomly divided into model group(M),Guizhi Fuling Capsule group(F),Yiqi Xiaozheng recipe high-dose group,medium dose group and low-dose group(G,Z,D).2.After 2 days of successful modeling,rats in each group were gavaged once a day for 4 weeks.During the experiment,the general state of rats was observed and weighed before modeling,2 and 4 weeks after modeling,and 2 and 4 weeks after drug intervention.3.After 4 weeks of administration,blood samples were collected from the heart of rats in each group to detect the indexes related to hemorheology and immune function;the uterus was removed,the weight of uterus was measured,and the uterine coefficient was calculated.Part of the tissues were fixed in 4% tissue cell fixed solution,and tissue sections were made for HE staining and immunofluorescence staining respectively;Some tissues were stored in 0.9% normal saline at room temperature,and the apoptosis and cycle distribution of cells were detected by flow cytometry within 2 hours;some tissues were stored in-80℃ refrigerator,and the protein expression levels of Bcl-2,Bax and caspase-3 were detected by Western blot.Result:1.General state of existence: there was no abnormality in the blank group.In the model group,there were anorexia,pale yellow and lusterless hair,depilation,irritability,mutual biting,etc.in the process of modeling,some rats had breast swelling and necrosis.In the process of drug intervention,the above symptoms in the model group were gradually aggravated;compared with the model group,the mental state of the rats in each treatment group was improved,their temperament tended to be calm,their hair color gradually became glossy,their hair removal alleviated,and their breast redness and swelling subsided,which was positively correlated with the administration time and concentration.2.Body weight and uterine coefficient: estrogen and progesterone load model,the weight of rats decreased significantly,uterine coefficient increased,compared with the blank group,there was a significant difference(P<0.01);drug intervention made the weight and uterine coefficient of rats improved significantly,the difference was statistically significant(P<0.05).However,Guizhi Fuling Capsule and Yiqi Xiaozheng prescription had the same effect on increasing body weight and decreasing uterine coefficient,and the difference was not statistically significant.3.Pathological changes of uterus: under the light microscope,the morphology of uterine smooth muscle cells in the blank group was normal,arranged orderly,and there was no hyperplasia and inflammatory cell infiltration in the myometrium;in the model group,the uterine smooth muscle was obviously thickened,the myometrium was arranged disorderly,with vacuoles,different cell sizes,and the glands were increased,and there were abundant capillaries and inflammatory cell infiltration in the thickened myometrium;in each treatment group,there were no significant differences The pathological changes of uterus were between the blank group and the model group.4.Hemorheological changes: the whole blood viscosity,plasma viscosity and hematocrit of the model group were significantly higher than those of the blank group(P<0.01).After 4 weeks of treatment,the blood viscosity and hematocrit of the treatment groups were significantly lower than those of the model group(P<0.01);the effect of group G was more significant than that of groups Z and D(P<0.05);the effect of group G and group F in improving blood stasis rheology was similar,and the difference was not statistically significant(P>0.05).5.Changes of immune function: The level of CD3+,especially CD4+,was significantly lower in the model group,while the level of CD8+ was significantly higher than that in the normal group(P<0.01).Each treatment group can significantly improve the immune function of rats,and can effectively regulate the changes of T cell subsets,with significant difference compared with the model group(P<0.05);The effect of Yiqi Xiaozheng recipe high dose group was better.6.Changes of estrogen and progesterone receptor expression in uterine tissue: compared with the blank group,the relative expression of ER and PR in uterine tissue of model group was significantly increased(P<0.05);in each treatment group,the inhibition of ER and PR expression in group G was more significant(P<0.05),but the effect of group D was not obvious.7.Apoptosis and cycle change of myoma cells:compared with the model,the apoptosis rate of myoma cells in each treatment group was significantly increased(P<0.01),but the effect of G group was more significant(P<0.05).Guizhi Fuling Capsule and Yiqi Xiaozheng prescription could inhibit the cells from resting phase to active phase,and significantly regulated the proportion of cells in G1/0 phase,S phase and G2 M phase,the difference was statistically significant(P<0.05).8.Changes of Bcl-2,Bax and caspase-3 protein expression in uterine tissue: In the model group,the expression of Bcl-2 protein was higher than that of Bax and caspase-3 protein(P<0.01).The expression levels of Bcl-2,Bax and caspase-3 were improved in all treatment groups,but the regulation effect of Yiqi Xiaozheng recipe was better than Guizhi Fuling Capsule(P<0.05).Conclusion:1.Yiqi Xiaozheng recipe can improve the general survival of rats with uterine fibroids.2.Yiqi Xiaozheng decoction can improve the pathological state of uterus in rats with uterine fibroids.3.Yiqi Xiaozheng recipe can regulate the immune function,promote blood circulation,reduce the high level of estrogen and progesterone in uterine tissue,inhibit the response of target cells to sex hormones,block cell cycle,promote the apoptosis of uterine smooth muscle cells,and maintain the dynamic balance of proliferation and apoptosis of smooth muscle cells,which may be one of the mechanisms of its treatment of uterine leiomyoma.