| ObjectiveIn this experiment,intramuscular injection of exogenous estrogen and progesterone was used to construct a rat uterine fibroids model to detect the expression level of transforming growth factors TGF-β3,BMP-2,and EMR markers leukemia inhibitory factor(LIF),homeobox gene A10(HOXA10)and homeobox gene A11(HOXA11).Explore the damage effect of UL on the EMR,and use the RCRS components compatibility to intervene in UL model rats,and study the RCRS components compatibility to improve the EMR in the UL environment.MethodsThe rat UL model was constructed by intramuscular injection of exogenous estrogen and progesterone.Forty-eight SD rats were randomly divided into 6 groups:blank control group(control group),model control group(model group),negative control group(GLX group),a combination of total flavonoids and zedoary turmeric oil group(RCRS group),total flavonoids of RC group and zedoary turmeric oil group(RS group),eight rats were arranged in each group(N=8).After a week’s adaptive feeding to rats,the model was established.The rats in the other groups were injected intramuscularly with 0.5 mg/kg body weight of estradiol benzoate injection into the inner thigh of the rat every morning,and the corpus luteum was intramuscularly injected into the other inner thigh of the rat every other day.The ketone injection is 1 mg·kg-1 body weight,0.5 mg kg-1(epinephrine hydrochloride)was injected subcutaneously into rats every 4 days.The above operation is continuous for 63 days.From the 36 th day,except for the control group and the model group,rats in the GLX control group,RCRS group,RC group and RS group were given GLX Capsules aqueous solution and RC total flavonoids-Zedoary volatile oil group every afternoon.Distribute the medicinal liquid,the medicinal liquid of total flavonoids of RS and the medicinal liquid of zedoary turmeric volatile oil,and the uninterrupted administration lasts for 28 days.(1)During the process of model building and administration of rats,record the weight and food intake of rats every week;(2)After modeling and administration,the rats are anesthetized,and blood is taken from the abdominal aorta to separate rat serum;(3)Take the rat uterus,ovary,heart,liver,spleen,lung,kidney and thymus to weigh,and calculate the organ coefficient of each main organ according to the wet weight of each organ,and then measure the rat uterus with a vernier caliper The horizontal and vertical diameters;(4)Calculate the coefficients of each major organ in the rat,perform HE staining on the uterine tissue to observe the pathological changes of the tissue,;(5)And use enzyme-linked immunosorbent assay(ELISA)to detect estrogen(E2),progesterone(P)and TGF-β3 expression level,;(6)Immunohistochemical technology(IHC)was used to detect the expression of estrogen receptor(ER),progesterone receptor(PR),BMP-2 and LIF in rat endometrial tissue;(7)And western blotting was used and fluorescence quantitative PCR were used to detect the protein expression and m RNA level of BMP-2,homeobox genes HOXA10/HOXA11 and TGF-β3.Results(1)The female rat UL model was successfully constructed: the weight of the control group rat rose steadily throughout the process,and the food intake fluctuated steadily.The color of the rat’s uterus was pale pink,the texture was soft and uniform,the uterine edge was straight,and there was no other appearance change.The pathological examination results showed that the uterine smooth muscle cells are tightly arranged,the tissue layer is uniform,and the outline is clear.The weight of the rats in the model group increased slowly,and there was no significant difference in food intake compared with the control group.The appearance of the uterus showed different degrees of swelling,dark color,deformity and edema.The horizontal and vertical diameters of the uterus increased significantly,and the uterine coefficient increased significantly.Pathological examination results showed that mucosal epithelial cell proliferation,ring necrosis of the endometrial ring,and a large number of inflammatory cell infiltrations were seen in the uterine tissue of the model group.The fibroids are staggered,and the leiomyoma cells in the lamina propria are actively proliferating and arranged disorder.The expression levels of E2,Pg and TGF-β3 in serum of model group rats were significantly higher than those of control group(p<0.05),and the difference was statistically significant,but the expression levels of estrogen receptor and progesterone receptor had no significant difference;(2)Influence of endometrial receptivity markers in rat UL model: After modeling,the test results showed that the expression level of LIF in rat endometrial tissue was down-regulated(p<0.05);the expression level of BMP-2 in endometrial tissue of model rats was significantly higher than that of control group rats(p<0.05),while the expression levels of HOXA10 and HOXA11 were significantly lower(p<0.05),with statistical significance;(3)The effect of the RC zedoary turmeric oil composition on the endometrial receptivity markers of the rat UL model: After UL rats are administered,the LIF expression level in the endometrial tissue of the RC group is higher than that of the model group(p<0.05),statistically significant;The expression level of BMP-2in the endometrial tissue of rats in the RCRS group,RC group,and RS group was significantly lower than that in the model group(p<0.05),which was statistically significant;In the endometrial tissue of the GLX group HOXA10 m RNA expression was significantly higher than that of model group rats(p<0.05),which was statistically significant.Compared with the model group,the expression of HOXA10 m RNA in rat endometrial tissues in the remaining three administration groups(RCRS group,S group,and E group)showed an overall downward trend although it was not statistically significant;The expression of HOXA11 m RNA in the endometrial tissue of rats in the RCRS,S,and E groups was significantly higher than that in the model group(p<0.05),which was statistically significant;(4)The influence of the zedoary turmeric oil composition on the TGF-β3/BMP-2 pathway of UL rat model: The level of TGF-β3 in the serum and endometrial tissue of UL rats increased significantly(p<0.05);The expression of BMP-2 in endometrium of model group was significantly higher than that of blank group(p<0.05).The expression level of TGF-β3 in rat endometrium of each administration group(RCRS group,RC group and RS group)was significantly lower than that of model group(p<0.05);and the expression level of BMP-2 in rat endometrium of RS group was significantly down-regulated(p<0.05).The levels of BMP-2 in the endometrium of rats in the other administration groups were lower than the expression of the endometrium in the model group,but there was no significant statistical significance,only showing a decreasing trend(p>0.05).Conclusion1.The method of exogenous intramuscular injection of estrogen and progesterone can build rat uterine fibroids,and can cause damage to rat EMR:markers of EMR in rat endometrial tissue The expression of HOXA10,HOXA11 and LIF increased,the expression level of BMP-2 decreased,and the expression level of TGF-β3 mediated by UL increased.2.Rhizome Sparganii total flavonoids composition can inhibit the excessive TGF-β3 level in rat endometrial tissue derived from UL,reduce the expression level of BMP-2 in rat endometrial tissue,and enhance the effect of rat endometrial on BMP on this basis,the levels of HOXA10,HOXA11 and LIF mediated by BMP-2 can be up-regulated,so as to improve the receptivity of the endometrium.3.The combination of RCRS group,Rhizome Sparganii total flavonoids and zedoary turmeric oil all have the effect of improving the endometrial receptivity of rats with uterine fibroids.In general,the role of the RCRS component compobility is due to the role of the single component. |
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