Experimental Study Of Electroacupuncture Combined With Granulocyte Colony-stimulating Factor (G-CSF) On Thin Endometrium In Rats

Objective:To investigate the effects of electroacupuncture and granulocyte colony-stimulating factor(G-CSF)on the endometrial receptivity of rats after treatment alone or in combination.Method :In the experimental study,50 female SD rats were randomly divided into five groups: normal group(N group),model group(M group),electroacupuncture group(E group),G-CSF group(G group),combined therapy.Group(F group).Model establishment method: In addition to group N,bilateral saline was infused with normal saline(NS)for 5 minutes,and other groups were perfused with absolute ethanol for 5minutes to establish a thin endometrial model caused by severe injury.Treatment groups of each group: 1N group,M group: free intake of feed and purified water,no intervention.Group 2E: After 3 weeks of model preparation,the electroacupuncture at "Sanyinjiao Point" and "Guanyuan Point" was treated for 3 weeks.3G-group: 5 weeks after model preparation,subcutaneous injection of G-CSF for 1 week.4F group: After 3weeks of model preparation,electroacupuncture “Sanyinjiao” and “Guanyuan acupoint”+ last week,subcutaneous injection of G-CSF.After the end of the treatment,the rats were sacrificed during the estrus period,HE staining of the endometrium was performed,and the morphology of the endometrium of each group was observed by electron microscope,and the endometrial thickness was measured.Immunohistochemistry(IHC)was used to detect the expression of endometrial vimentin(vim)and cytokeratin(CK-19).PCR detection of endometrial receptivity related indicators: vascular endothelial growth factor(VEGF),integrin ???3,homeobox gene 10(HOXA10)m RNA expression;PCR detection of intimal profibrotic factors: transforming growth factor-?1m RNA expression level;PCR detection of intimal proinflammatory factors: tumor necrosis factor(TNF)-?,interferon(IFN)-?m RNA expression.Result:The results of pre-experimental endometrial pathology showed that the morphology of endometrium in the normal group was significantly better than that in the model group.The endometrial thickness of the model group was significantly lower than that of the normal group(P<0.01).Pathological results 1:1 comparison of endometrial thickness in each group:The intima thickness of group M was significantly lower than the other groups(P<0.01);in each treatment group,the average intima thickness of group E was the highest.Pathological results 2:1 Comparison of vim positive area in endometrium of each group: N group and E group were significantly higher than the other groups(P<0.05);G group was significantly smaller than M group(P<0.05);The intimal vim positive area was the lowest;2 the endometrial CK-19 positive area of each group was compared: the other groups were significantly lower than the other groups(P<0.05);the other groups were not significantly different.The difference was(P>0.05).The average expression of CK-19 in the endometrium of group E was the largest.Endometrial receptivity related indicators: 1 The expression of VEGF m RNA in the endometrium of each group: M group was significantly lower than N group,E group and F group(P<0.01);G group was significantly lower than E group(P<0.01)P<0.01);the average VEGF m RNA expression of the endometrium in group E was the highest.2 The m RNA expression levels of endometrial integrin ???3 in each group were significantly lower in group M and group F than in group N(P<0.05);group E and group G were significantly higher than group M;group E endometrial integrin The m RNA expression level of ???3 is the highest.The m RNA expression of Hox A10 in the endometrium of each group was significantly higher than that in the other groups(P<0.01).There was no significant difference between the other groups(P>0.05).The results of endometrial profibrotic factors: 1 The expression of TGF-?1m RNA in each group: Compared with the other groups,the expression of the other groups was significantly lower than that of the N group(P<0.01);The average TGF-?1 m RNA expression level was the lowest.The results of endometrial pro-inflammatory factors: 1 The expression of endometrial(TNF)-? m RNA in each group: There was no significant difference between the groups(P>0.05);the average of endometrium in group G(The m RNA expression level of TNF)-? was the lowest.2 The m RNA expression levels of each group(IFN)-? were compared: the other groups were significantly higher than the other groups(P<0.05);the M group was significantly higher than the treatment groups(P<0.05);The m RNA expression of endometrium(IFN)-? in group F was the lowest.in conclusion:A.The pathological results of endometrium in the pre-experimental model group were characterized by thin endometrium,suggesting that the rat model of injury-type thin endometrium was successfully established.B.Electroacupuncture and G-CSF alone or in combination with the thin endometrium of rats can improve the thickness and morphology of thin endometrium,but there is no significant difference between the combination therapy and the treatment alone.C.The electroacupuncture group significantly increased the thickness of the thin endometrium,which may be related to the promotion of the expression of VIM,VEGF,integrin ???3 and HOXA10 in the endometrium and the inhibition of the expression of proinflammatory cytokine(IFN)-?.D.The G-CSF group significantly increased the thickness of the thin endometrium,which may be related to the promotion of the expression of endometrial integrin???3 and the inhibition of the expression of proinflammatory factor(IFN)-?.E.The combination group significantly increased the thickness of the thin endometrium,which may be related to the promotion of endometrial VEGF expression and inhibition of proinflammatory factor(IFN)-? expression.