| Objects: In order to clarify that deficiency of kidney and real of liver is thebasic pathogenesis of immunity infertility theoretically, this paper establishedimmunity infertility disease models with the positive male mice, active immunizationof homologous modified character method was used, studied the effect ofTiaomianyulitang on parameters of semen, testis constitution structure, acidphosphatase, apoptosis of spermatogenic cells, mouse testis bcl-2 & bax expressions,anti-sperm antibody reversing ratio, and pregnancy rate of matching female mice inmodel mice, revealed the pathological mechanism of immunity infertility and Therapymechanism of Tiaomianyulintang.Methods:①Totally 120 male mice at the age of three months, ten normal micewere randomly selected as normal control group, another sixty mice were separatelyinjected antigen to get immune around subcutaneous lymph nodes of bilateral inguinaland both sides of neck and back spine subcutaneous, together six parts respectively.Booster immunizations were employed at ten-day interval upon such parts above. Tenml blood was adopted from tails before the first immunization and after every timeintensify immune, then separated serum and detected anti-sperm antibody of serum byenzyme linked immune sorbet assay. After immunizations for four times, fifty-sevenrats' serum was positive and these mice were randomly divided into model group byrandom number table method, prednisone acetae group, Tiaomianyulintang low dosegroup, Tiaomianyulintang middle dose group, Tiaomianyulintang high dose group,which were separately given oral medicine. Normal group and model group wereadministered with physiologic saline orally, each group of Tiaomianyulintang wasgiven decoction of Tiaomianyulintang, prednisone acetae group was given solution ofprednisone acetae, once a day. The procedures lasted for 30 days.②Detecting theeffect of Tiaomianyulintang on parameters of semen of mice, and detecting thehistological changes of testes.③Detecting activity of rat's semen with acidphosphatase, and determining the effect of Tiaomianyulinrang on activity ofimmuneosuppressivein testis of model mice.④Detecting apoptosis ofspermatogenic cells of mice in each group by flow cytometer.⑤Detectingexpressions of bcl-2, bax gene combine with testis tissue apoptosis of male mice ineach group by immunohistochemistry.⑥To observe the effect of Tiaomianyulintangon anti-sperm antibody reversing ratio and to collect group statistics of the number ofembryos and pregnancy rate after stopping drug when female mice were added to themale in each group.Results:①Changes of parameters of semen in model mice. Parameters ofsemen such as sperm density, sperm motility rate, speed of sperm motion straightlineand so on decreased rapidly in model mice, significantly different from normal controlgroup and other therapy groups. Parameters of semen of male mice in prednisoneacetae group differed greatly from those of Tiaomianyulintang middle dose group and Tiaomianyulintang high dose group.②Micro and ultramicroformation of testistissue. In model group, atrophy of the spermatogenic epithelium in mice was observedunder microscope, epithelium inside lumen became thinner, some spermatogenicnucleus pyknotic degeneration, interstitial edama and some stromal cells vacuolardegeneration were found in it. In each Tiaomianyulintang group and prednisoneacetae group, spermatogenic cells at all levels of testis issues increased, structural ofseminiferous epithelium became more complete, and the lesion of testis issues wererepaired in various degree, among which the effects of Tiaomianyulintang middledose group and high group were better than that of prednisone acetae group. Throughtransmission electron microscope, the apoptosis of rat stertoli cells and sperm cells atall levels in model group were servely injured. There were a great deal of vacuoles incytoplasm of stertoli cells, gaps of sperm cells at all levels broadening, mitochondrialswelling and spermatid uncommon. Therapy groups had renovate effect on thedamage of the apoptosis of mouse stertoli cells and sperm cells at all levels,Tiaomianyulintang middle andhigh groups alleviated the damaged degree noticeably.③Changes of acid phosphatase activity of testis immunosuppressive in model malemice. Acid phosphatase activity in testicular tissue homogenate of mice in modelgroup went down notably, significantly different from the normal control group. Theacid phosphatase activity of prednisone acetae group had great difference toTiaomianyulinrang middle and high dose groups. There was no noticeable differencebetween Tiaomianyulintang middle and high groups.④The apoptosis number andmouse of testicular spermatogenic cells in mice of model group. There were manyspermatogenic cell apoptosis in model group, and the apoptosis number and rate oftesticular spermatogenic cells in prednisone acetae group were much lower than thosein model group. There was no great difference between Tiaomianyulinrang low groupand prednisone acetae group. There was significant difference betweenTiaomianyulintang middle and high dose groups and prednisone acetae group.⑤Expression of gene correlated with testicular tissue apoptosis of mice in model group.Compared with the normal group, the expression of bcl-2 in testicular tissue of micein model group decreased, and bax's expression increased. Compared with modelgroup, the expression of bcl-2 improved obviously and that of bax decreased notablyin Tiaomianyulintang middle and high dose groups.⑥Statistics of anti-spermantibody reversing ratio in each group for vaginal plug and pregnancy rate of femalemice when they were mated with male mice. After therapy for thirty days, there wasobviously difference between Tiaomianyulinrang groups and prednisone acetae group.Anti-sperm antibody reversing ratio in Tiaomianyulintang groups were very close.The result showed that the statistics of pregnancy rate and embryos number of eachpregnancy of Tiaomianyulintang groups were obviously difference to model group,and also to prednisone acetae group.Conclusion: This paper brought forward a point of view that deficiency ofkidney and real of liver is the basic pathogenesis of immunity infertility, and clarifypathogenesis of immunity infertility based on traditional chinese medicine basic theory. Through these experiments, we found that Tiaomianyulintang could enhancesperm density, sperm motility rate, speed of sperm motion straightline in model micesignificantly and repair the lesion of testis issues successfully. High dose groupincreased obviously acid phosphatase activity in testicular tissue homogenate of micein model group and immune function of male mice were ameliorated markedly.Middle and high dose groups decreased the apoptosis rate of testicular spermatogeniccells in mice of model group, and increased the number of spermatogenic cell at alllevels. In addition, it could upregulate the expression of bcl-2 which gene inhibitingapoptosis and downregulate the expression of bax which gene accelerating apoptosis,and it could made anti-sperm antibody in model mice turned negative. Besides, thepregnancy rate of female mice and the number of embryos in Tiaomianyulintanggroup were both higher than prednisone acetae group and model group.The basic pathogenesis of immunity infertility lies in deficiency of kidney andreal of liver, and deficiencyexcess miscellaneous is the character. Based on theprinciple of Neijing that reinforcing deficiency and reducing excess.TheStandardization is Replenishing Qi and Nourishing Kidney Deficiency, Clearing Heatand Removing Damplaw, Blood-activatin and Stasis-eliminating, Treating liver andkidney and using Reinforcing-Reducing Method at the same time all these has a bettereffect on treatment of immune system confusion. Tiaomianyulitang turned anti-spermantibody negative in model immune infertility mice, and enhanced obviouslyreproductive capability of these mice. These effects revealed the scientific of thetheory that deficiency of kidney and real of liver is the basic pathogenesis ofimmunity infertility.
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