|
Endometriosis (Endometriosis, EMS, referred to endometriosis) is a chronic gynecological etiology is complex and is closely related with cancer, the World Health Organization (WHO) as one of incurable diseases. Current treatment of this disease with hormone drugs, based on surgery. Since long-term hormone therapy accompanying false pregnancy, false menopause symptoms, and indications for surgical treatment limitations and complications arising, physical and mental health to patients caused many adverse effects. According to Chinese medicine dialectical theory of governance, in relieving symptoms, such as menstruation progesterone has its unique effects and toxicity, long-term use. So look for reliable efficacy of traditional Chinese medicine preparations, and to explore its mechanism of action has important research value. Tri pill was first seen in \Ectopic endometrial cells grow outside the uterine cavity and survived, with its spontaneous apoptosis weakened or not sensitive to apoptotic signals related to changes in other characteristics. This topic from the perspective of apoptosis observed blood circulation triangular square flavored pill main ingredient (referred SLW) endometriosis treatment efficacy and to explore its mechanism. Objective To observe the Tri-flavored pill main ingredient for endometriosis rat ectopic endometrial development and apoptosis; observation flavored pill main ingredient against Tri-apoptotic gene Bcl-2 mRNA and induces cell wither the death of the role of Fas mRNA genes. Explore the main ingredient Modified triangular pill endometriosis treatment efficacy and induction of ectopic endometrial cell apoptosis. Method: 1, animal model selected 50 healthy female Wistar rats, weighing 180 ~ 220g, regular daily observation of vaginal smears, select the estrous cycle of 4 to 5 days and there are two consecutive estrous cycles of female rats at estrus (vaginal smears of keratinized epithelial cells) line modeling surgery. 5% chloral hydrate (0.7ml/100g) anesthetized by intraperitoneal injection, routine disinfection, under sterile conditions laparotomy, ligation of the right uterine ends after clipping about 1.5 cm from the distal segment of the uterus, removing fat tissue, cut 5mm × 5mm uterine tissue 3. One sent for pathological examination (confirmed as uterine tissue) and another two near the peritoneal vessels were sewn in the abdominal wall to the mucosal surface facing the peritoneum, serosal side facing the abdominal cavity, the abdomen was closed. Each mouse by intraperitoneal injection of gentamicin 0.1ml × 3 days. Normal cleaning feeding. 2, the models incorporate standard modeling after four weeks, and vaginal cytology smears, the second laparotomy observe the growth of endometriosis and transplanted with a vernier caliper measuring tumor size. If the migration stove volume increases, the volume ≥ 8mm3, were uplifted translucent nodular, cystic, or omentum covered by connective tissue and blood vessels to form, there are light yellow transparent cyst fluid accumulation. Cut transplant stove inspected, confirmed transplantation stove in endometrial epithelial cells, glands and stroma growth. 3, the test group and medication in addition to the normal control group, and the other will be successful model rats were randomly divided into SLW 0.11mg.kg-1.d-1 group, 0.34mg.kg-1.d-1 group, 1.02mg . kg-1.d-1 group, 0.5mg.kg-1.d-1 Gestrinone group, model control group, administered orally once a day for 4 weeks. 4, four weeks after the end of the detection index treatment, observation of vaginal cytology smears rats were killed after the third laparotomy observed ectopic endometrial growth, and to measure the transplanted tumor volume. Cut eutopic and ectopic endometrium, HE staining morphology pathology prosecution. TUNEL assay ectopic endometrial cells, in situ hybridization apoptosis genes Bcl-2 and Fas mRNA expression. Results: 1, SLW rat ectopic endometrial tissue growth medication for 4 weeks after transplantation tumor model control group size slightly larger than four weeks ago, was a small bulge translucent bubble, the surface blood vessels clearly visible, filled with fluid inside . And Gestrinone group and SLW 0.11mg.kg-1.d-1 group, 0.34mg.kg-1.d-1 group, 1.02mg.kg-1.d-1 xenograft tumor volume was significantly reduced, cystic bubble surface color bleak, individual transplant stove color yellow, showing atrophy shape. 2, SLW rat ectopic endometrial tissue microstructure HE staining, light microscope. Survival of transplanted endometrial tissue, and the resulting respectively from inside to outside wall epithelial layer, stromal cell layer and a small amount of fibrous connective tissue capsule a small amount of white blood cells and mucus floating. Model group ectopic endometrial glandular epithelial cells showed a high or low columnar columnar, arranged in a vesicle lumen surface, closer connections between cells, the nucleus large, oval, located at the base, abundant cytoplasm. Stroma is also rich in individual parts of which form the epithelial invagination false glands. SLW1.02mg.kg-1.d-1 group, 0.34mg.kg-1.d-1 group and Gestrinone group, endometrial glandular epithelium was thinner, even short columnar cells were flat, loosely arranged , irregular nuclei, location uncertain, basophilic cytoplasm increases. Stroma was significantly reduced or absent fewer glands. 3, SLW rat ectopic endometrial tissue volume effects before treatment, the rats endometriotic lesion volume no statistical significant difference (P gt; 0.05). After 4 weeks, Gestrinone group and SLW1.02mg.kg-1.d-1 group, 0.34mg.kg-1.d-1 group, 0.11mg.kg-1.d-1 group within ectopic membrane volume was 17.01mm3, 18.42mm3, 25.90mm3, 58.34mm3, volume was significantly reduced, compared with the model group 102.17mm3 were significant differences (P lt; 0.05). The model control group ectopic endometrium than four weeks before the volume increased slightly, but no significant difference (P gt; 0.05). The results showed: Modified triangular pills can significantly reduce the volume of ectopic endometrium. 4, SLW rat ectopic endometrial cell apoptosis after 4 weeks of treatment, ectopic endometrial tissue sections under a fluorescence microscope to 520 ± 20nm wavelength observation shows that the marked green fluorescent apoptotic cells. Confocal laser scanning, display apoptotic nuclei lysis, apoptotic bodies (apoptotic cells characteristic morphological changes) form. Model control group, the average fluorescence intensity than the normal control group 0.1892 0.1457 significantly lower (P lt; 0.05), model group decreased ectopic endometrial cells, growth imbalance. And Gestrinone group and SLW1.02mg.kg-1.d-1 group, 0.34mg.kg-1.d-1 group, 0.11mg.kg-1.d-1 group, the average fluorescence intensity values ??were 0.2315 , 0.2246,0.2209,0.1955, 0.1457, compared with the model group, the mean fluorescence intensity values ??were significantly increased (P lt; 0.05). The results showed: Modified triangular pill can promote ectopic endometrial cells. 5, SLW ectopic endometrial tissue of rat Bcl-2 mRNA expression after 4 weeks of treatment, in situ hybridization showed that anti-apoptotic gene Bcl-2 mRNA positive cells stained brown, mainly in the gland epithelial cells and stromal cell cytoplasm, membrane and nuclear membrane. Model control group ectopic endometrial tissue Bcl-2 mRNA expression was positive, and the majority of strong expression, the average optical density value 0.1728 0.1491 compared with the control group there was significant difference (P lt; 0.05). Gestrinone group and SLW1.02mg.kg-1.d-1 group, 0.34mg.kg-1.d-1 group, 0.11mg.kg-1.d-1 ectopic endometrial tissue Bcl-2 mRNA expression compared with the model group was significantly reduced, the average optical density values ??were 0.1014,0.1055,0.1102,0.1370, 0.1728 compared with the model group were significantly different (P lt; 0.05). The results showed: Modified triangular pill can reduce the ectopic endometrial tissue anti-apoptotic gene Bcl-2 mRNA expression, thus promoting ectopic endometrial cells. 6, SLW rat Fas mRNA expression in ectopic endometrial tissue after 4 weeks of treatment, in situ hybridization showed that apoptosis gene Fas mRNA positive cells stained brown, mainly in endometrial stromal cells cytoplasm and cell membrane. Model control group ectopic endometrial tissue expression of Fas mRNA only a small amount, and the lighter staining, the mean optical density value 0.0883 0.0912 compared with normal control group had no significant difference (P gt; 0.05). And each treatment group ectopic endometrial tissue expression of Fas mRNA compared with the model group increased in varying degrees, Gestrinone group and SLW1.02mg.kg-1.d-1 group, 0.34mg.kg-1. d-1 group, 0.11mg.kg-1.d-1 group, the average optical density (0.1242,0.1206,0.1164,0.0966) with the model group (0.0883), the difference was statistically significant (P lt; 0.05). The results showed: Modified triangular pill can increase the ectopic endometrial tissue apoptosis gene Fas mRNA expression, thereby inducing ectopic endometrial cells. Conclusions: 1, flavored triangular pill inhibits rat endometriosis development, reduction of ectopic endometrial volume and improve the microstructure ectopic endometrial pathology, and promote ectopic endometrial cells. 2, flavored triangular pill rat endometriosis development mechanism, the inhibition of ectopic endometrial tissue anti-apoptotic gene Bcl-2 mRNA expression increased apoptosis in Fas mRNA gene expression, thereby inducing ectopic endometrium is closely related to apoptosis.
|