| Objective: The intrauterine inflammatory microenvironment composed of inflammatory cells and their factors is an important part of the concept of endometrial receptivity.This experiment was to observe the morphological changes of endometrium,expression levels of LXA4,IL-1α,IL-1β,IL-18,IL-4,TGF-β and LIF in the implantation window of rats with kidney deficiency and blood stasis syndrome.Preliminary this paper discusses on the mechanism of to kidney-supplementing and blood-moving prescription by rectal administration improve endometrial receptivity by regulating the uterine inflammatory microenvironment and embryo-intimal interaction in the implantation window.Methods: The adult unpregnant female SD mouse that in proestrus were randomly divided into six groups,namely blank group,model group,high dose group of kidney-supplementing and blood-moving prescription,low dose group of kidney-supplementing and blood-moving prescription,Chinese medicine control group and Aspirin control group,by vaginal smear and determination.In addition to the model group,the remaining groups lavage by hydroxyurea orally plus adrenaline hypodermic cause kidney deficiency and blood stasis composite model.At the same time modeling of kidney deficiency and blood stasis,each group was given an intervention drug.When the kidney deficiency and blood stasis model is established,the female rats in estrus in each group were screened by the vaginal secretion,male and female rats by 2: 1 ratio to be mated.Referred to as first day of pregnancy if it was found vaginal smears observed sperm the second day,and then stop treating pregnant rats administered.On the fourth day of pregnancy,rats were injected subcutaneously with 5.5 mg/kg of mifepristone solvent.On the fifth day,the abdominal aorta blood was taken,centrifuged,and the serum was separated and stored at-20 ° C for examination;After weighing the bilateral uterus,take the appropriate amount of uterine tissue on the same side to make tissue homogenate,and store at-20 °C for testing.The remaining uterus was placed in 4% formaldehyde solution for 48 h to make pathological sections.Observing uterine HE stained sections and immunohistochemical sections by optical microscope.The expressions of LXA4,IL-1α,IL-1β,IL-18,IL-4,TGF-β and LIF in uterine tissue were detected by ELISA.The content of E2 and P in serum was detected by ELISA.Results: 1.Effect of kidney-supplementing and blood-moving prescription by rectal administration on Expression of E2 and P in the implantation window of Rats in different groups The serum E2 content of the model group was lower than that of the blank group(P<0.05).The serum E2 content in the drug-high group and the Chinese medicine control group was significantly higher than that in the model group(P<0.001).Compared with the blank group,the serum P content in the model group significantly decreased(P<0.001).Compared with the model group,the serum P level in the drug high group and the Chinese medicine control group increased significantly(P< 0.05)2.Effect of kidney-supplementing and blood-moving prescription by rectal administration on morphology of uterine in the implantation window of Rats in different groups In the model group,the endometrial epithelium was very thin,the number of epithelial cells was small,the interstitial was dense,and the glands and spiral arterioles were less.The blank epithelium was thicker in the endometrium,and the number of epithelial cells was large and the vacuoles were visible under the nucleus.The interstitial was loose and uniform,the glands were visible and regular,and the spiral arterioles were abundant.The coated epithelium in the low and high doses of the drug had a different degree of growth than the model group,and the number of epithelial cells was higher(the vacuole was observed in the high drug group).The interstitial is looser and more uniform,with more glands and more spiral arterioles. The overall situation of the Chinese medicine control group and the western medicine control group is between the low drug and high drug groups.4.Effect of kidney-supplementing and blood-moving prescription by rectal administration on expression of TGF-β in Uterus Tissue of rats in different groups Under light microscopy,it was found that TGF-β protein was expressed in the cytoplasm of epithelial cells,mesenchymal cells and glandular cells of rat endometrium,and its positive characteristics were yellow particles with different shades.The TGF-β in the blank group showed a yellow-brown color,a high density,and a wide coverage area.In contrast to the model group,it appears light yellow,with low density and low coverage.The remaining four groups performed in color,density,and area between the first two groups.5.Effect of kidney-supplementing and blood-moving prescription by rectal administration on proinflammatory cytokines IL-1α,IL-β and IL-18 in uterine microenvironment of rats with kidney deficiency and blood stasis during the implantation window in different groups IL-1α expression in the blank group,the low-dose group,and the high-dose group increased compared with the model group(P<0.05);IL-1β expression in the blank group,the low-dose group,and the high-dose group was lower than that in the model group(P<0.05);the expression of IL-1β in the aspirin control group was increased compared with the high-dose group.IL-18 expression in the blank group,the high-dose group and the aspirin control group was increased compared with the model group(P<0.05).6.Effect of kidney-supplementing and blood-moving prescription by rectal administration on anti-inflammatory cytokines IL-4,TGF-βand LIF in uterine microenvironment of rats in different groups Compared with the model group,the expression of IL-4 was increased in the blank group and the Chinese medicine control group(P<0.05).The expression of IL-4 in the high-dose group was increased compared with the model group,and the difference was significant(P<0.001).The expression levels of IL-4 in the model group,the drug low group,the Chinese medicine control group and the aspirin control group were lower than those in the drug high group(P<0.05).The expression levels of TGF-β and LIF in the blank group,high-dose group and Chinese medicine control group were increased compared with the model group(P<0.05).7.Effect of kidney-supplementing and blood-moving prescription by rectal administration on balance of inflammatory cytokines in uterine microenvironment of rats in different groups The ratio of IL-1β/IL-4 in the blank group,low-dose group,high-dose group,Chinese medicine control group and aspirin group was significantly different from that in the model group(P<0.05).When the ratio of L-1α/TGF-β was compared between the model group and the high-dose group,the difference was statistically significant(P<0.05).The ratio of IL-1β/ TGF-β in the blank group,the low dose group,the high dose group and the Chinese medicine control group was statistical different from that in the model group(P<0.05).The IL-1β/ LIF ratios of the blank,low-dose,high-dose,and traditional Chinese medicine control groups were statistically different compared with the model(P<0.05).Conclusion: 1.Kidney-supplementing and blood-moving formula by rectal administration improves the endometrial receptivity by up-regulating the expression of steroid hormone estradiol and progesterone in the implantation window of rats with kidney deficiency and blood stasis,thus helping embryo implantation.2.Kidney-supplementing and blood-moving formula by rectal administration may regulate the balance of proinflammatory and anti-inflammatory factors in the microenvironment of uterine tissue at the implantation window of rats with kidney deficiency and blood stasis,which improves embryonic-endometrial dialogue exchange and endometrial receptivity,and improves the success rate of embryo implantation. |
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