加味三棱丸对大鼠异位内膜细胞凋亡及其机制的研究
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摘要
子宫内膜异位症(Endometriosis,EMS,简称内异症)是妇科顽症,病因复杂,且与恶性肿瘤密切相关,被世界卫生组织(WHO)列为疑难病症之一。目前治疗本病多以激素类药物、手术为主。由于长期激素治疗伴随的假孕、假绝经现象,以及手术治疗适应症的限制和并发症的产生,给患者身体和心理健康造成了许多不良影响。中医药根据辩证论治,在缓解临床症状、调经助孕等方面有其独特疗效,且毒副作用小,可长期服用。因此寻找疗效可靠的中药制剂,并探讨其作用机理具有重要的研究价值。三棱丸最早见于《经验良方》,是由三棱、莪术两味活血化瘀中药组成,用于瘀血经闭,行经腹痛,癥瘕积聚,食积腹痛。异位子宫内膜细胞在宫腔外种植并存活,与其自发性凋亡减弱或对凋亡信号不敏感等特性改变有关。本课题从细胞凋亡的角度观察活血化瘀方加味三棱丸主要成份(简称SLW)治疗子宫内膜异位症的疗效并探讨其作用机制。
目的观察加味三棱丸主要成分对子宫内膜异位症大鼠异位内膜的发展及其细胞凋亡的影响;观察加味三棱丸主要成分对抗凋亡基因Bcl-2 mRNA和诱导细胞凋亡的基因Fas mRNA的作用。探讨加味三棱丸主要成分治疗子宫内膜异位症的疗效及诱导异位内膜细胞凋亡的机制。
方法:
1、动物模型的制作
选取健康雌性Wistar大鼠50只,体重180～220g,每天定时观察阴道细胞涂片,选择动情周期4～5天并连续有2个动情周期的雌性大鼠,于动情期(阴道细胞涂片为角化上皮细胞时)行造模手术。以5%水合氯醛(0.7ml/100g)腹腔注射麻醉,常规消毒,在无菌条件下开腹,结扎右侧子宫两端后,从远端剪取长约1.5㎝的子宫段,剔除脂肪组织,剪成5mm×5mm的子宫组织3块。一块送病理检验(确证为子宫组织),另外两块靠近腹膜血管处,缝于腹壁,以粘膜面朝向腹膜,浆膜面朝向腹腔,关腹。每鼠腹腔注射庆大霉素0.1ml×3天。正常清洁饲养。
2、模型纳入的标准
建模后4周,观察阴道细胞学涂片,第二次剖腹观察异位子宫内膜的生长情况,并用游标卡尺测量移植灶大小。如果移植灶体积增大,体积≥8mm3,呈隆起透亮的结节状、囊状,被结缔组织或大网膜覆盖并有血管形成,内有淡黄色透明囊液积聚。切取移植灶送检,证实移植灶中有子宫内膜上皮细胞、腺体及间质的生长。
3、试验分组与用药
除正常对照组外,另将造模成功的大鼠随机分为SLW 0.11mg.kg-1.d-1组、0.34mg.kg-1.d-1组、1.02mg.kg-1.d-1组、0.5mg.kg-1.d-1孕三烯酮组、模型对照组,灌胃给药,每天1次,连续4周。
4、检测指标
4周后结束治疗,观察阴道细胞学涂片后处死大鼠,第三次剖腹观察异位内膜生长情况,并测量移植灶体积。切取在位子宫与异位子宫内膜, HE染色进行形态学病理检察。TUNEL法检测异位内膜细胞凋亡,原位杂交检测凋亡调控基因Bcl-2和Fas mRNA的表达。
结果:
1、SLW对大鼠异位内膜组织生长的影响
用药4周后,模型对照组移植灶体积比4周前略大,呈隆起透亮的小泡状,表面血管清晰可见,内部充满积液。而孕三烯酮组和SLW 0.11mg.kg-1.d-1组、0.34mg.kg-1.d-1组、1.02mg.kg-1.d-1组移植灶体积明显缩小,囊泡表面颜色黯淡,个别移植灶颜色泛黄,呈现萎缩状。
2、SLW对大鼠异位内膜组织显微结构的影响
HE染色,光镜下观察。移植的子宫内膜组织成活,形成的囊壁由内向外分别为上皮细胞层、间质细胞层及少量纤维结缔组织,囊内为少量漂浮的白细胞和黏液。模型对照组异位内膜腺上皮细胞呈高柱状或矮柱状,排列在囊泡内腔面,细胞间连接较紧密,细胞核大,呈椭圆形,位于基底部,胞浆丰富。内膜间质也丰富,其中有个别部位上皮层内陷形成假腺体。SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组和孕三烯酮组,内膜腺上皮层明显变薄,细胞呈矮柱状甚至扁平状,松散排列,核不规则,位置不定,胞浆嗜碱性增加。内膜间质也明显减少,腺体少甚至缺失。
3、SLW对大鼠异位内膜组织体积的影响
用药前,各组大鼠异位内膜病灶体积经统计学处理无显著性差异(P >0.05)。用药4周后,孕三烯酮组和SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组、0.11mg.kg-1.d-1组异位内膜体积分别为17.01mm3、18.42mm3、25.90mm3、58.34mm3,体积明显缩小,与模型对照组102.17mm3相比均有显著性差异(P < 0.05)。而模型对照组异位内膜体积比4周前略有增加,但无显著性差异(P >0.05)。结果显示:加味三棱丸可显著缩小异位内膜体积。
4、SLW对大鼠异位内膜细胞凋亡的影响
用药4周后,异位内膜组织切片在荧光显微镜下,以520±20nm波长观察,可见绿色荧光所标记的凋亡细胞。共聚焦激光扫描,显示凋亡细胞核裂解,凋亡小体(凋亡细胞特征性的形态学改变)形成。模型对照组平均荧光强度值0.1457比正常对照组0.1892显著降低(P <0.05),模型对照组异位内膜细胞凋亡减少,生长失衡。而孕三烯酮组和SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组、0.11mg.kg-1.d-1组平均荧光强度值分别为0.2315、0.2246、0.2209、0.1955,与模型对照组0.1457相比,其平均荧光强度值均显著增高(P <0.05)。结果显示:加味三棱丸能促进异位内膜细胞凋亡。
5、SLW对大鼠异位内膜组织Bcl-2 mRNA表达的影响
用药4周后,原位杂交结果显示,抗凋亡基因Bcl-2 mRNA阳性表达的细胞染色为棕黄色,主要分布在腺上皮细胞和间质细胞的胞浆、胞膜和核膜。模型对照组异位内膜组织Bcl-2 mRNA呈阳性表达,且多数为强阳性表达,其平均光密度值0.1728与正常对照组0.1491相比有显著性差异(P <0.05)。孕三烯酮组和SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组、0.11mg.kg-1.d-1组异位内膜组织Bcl-2 mRNA的表达较模型对照组显著减少,其平均光密度值分别为0.1014、0.1055、0.1102、0.1370,与模型对照组0.1728比较均有显著性差异(P <0.05)。结果显示:加味三棱丸能降低异位内膜组织抗凋亡基因Bcl-2 mRNA的表达,从而促进异位内膜细胞凋亡。
6、SLW对大鼠异位内膜组织Fas mRNA表达的影响
用药4周后,原位杂交结果显示,凋亡基因Fas mRNA阳性表达的细胞染色为棕黄色,主要分布在内膜间质细胞的胞浆、胞膜。模型对照组异位内膜组织Fas mRNA仅有少量表达,且染色较浅,其平均光密度值0.0883与正常对照组0.0912相比无显著性差异(P >0.05)。而各治疗组异位内膜组织Fas mRNA的表达均较模型对照组有不同程度的增加,孕三烯酮组和SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组、0.11mg.kg-1.d-1组平均光密度值(0.1242、0.1206、0.1164、0.0966)与模型对照组(0.0883)比较,差异有显著性(P <0.05)。结果显示:加味三棱丸能增加异位内膜组织凋亡基因Fas mRNA的表达,从而诱导异位内膜细胞凋亡。
结论:
1、加味三棱丸可抑制大鼠子宫内膜异位症的发展,缩小异位内膜体积,改善异位内膜病理学显微结构,促进异位内膜细胞凋亡。
2、加味三棱丸抑制大鼠子宫内膜异位症发展的机制,与其抑制异位内膜组织抗凋亡基因Bcl-2 mRNA表达、增加凋亡基因Fas mRNA表达,从而诱导异位内膜细胞凋亡有密切相关。
AIM
The model of endometriosis in Wistar rats by transplantation were established to investigate effects of traditional Chinese medicine compounds Sanleng pellet (SLW) on the volume of the ectopic endometrial tissue and the apoptosis of endometriotic cells. The effects of SLW on Endometriosis in Wistar rats were investigated in the expression of Bcl-2 mRNA and Fas mRNA in ectopic endometriotic tissue, meanwhile study the mechanism of SLW’s apoptosis induction in ectopic endometria.
METHOD
50 female wistar rats, 180-220 grammar, were selected to observe the smear of vaginal cell at settled time every day. Those rats whose estrus cycle is 4-5 days and which has continuous 2 estrus cycle were selected to establish rat models with endometriosis during estrus (the smear of vaginal cell is cuticular epithelial cells) according to Vernon′s method.
Four weeks after the autotransplantation, the rats were paunched the second time to investigate the growth of transplantation and mesure its volume. Those rats whose autotransplantation was not well growth were eliminated. The model rats with peritoneal implants were randomly assigned into five groups: the model control group and four treatment groups which were given respectively normal sodium, high-dose of SLW, moderate-dose of SLW, low-dose of SLW and gestrinone served as the control drugs continuously for four weeks. Meanwhile, eight normal rats were used as the blank control.
After four weeks treatment, the model rats were killed after observing the smear of vaginal cell. The rats were paunched the third time to observe the shape and mesure the volume of ectopic endometrial tissues. Eutopic endometrium and ectopic implants were cut, dipped in 4% paraform, paraffin embed, 6μm thick continuous slice made and HE stained to observe the pathologic change of eutopic and ectopic endometrium. Apoptosis in ectopic endometria was detected by TUNEL method. The score of the mRNA expression levels of Bcl-2 and Fas of ectopic implants were detected by in-situ hybridization test. Every slice was observed five fields of vision randomly under microscope. Mean fluorescence intensity and average optical density value were dealed with the image analytical software.
All data were showed by x±SD and dealed with SPSS11.5 statistical software. It is statistical significance when P<0.05.
RESULT
1. Effect of SLW on the volume size and morphological change of heterotopic tissue of EMS rats
Before treatment, the volume size value of endometrial implants has not significant diference between each treatment group and the model control group (P >0.05). After four weeks’treatment, the positive control group and SLW different doses groups could reduce the volume on the different degree, SLW high and middle dosage have the striking effect. Compared with endometriosis models, the high, middle and low dose groups could remarkably reduce heterotopic tissue volum size from 102.17±58.27mm3 to 18.42±13.21mm3, 25.90±15.26mm3and 58.34±37.05mm3 (P <0.05).
Under the light microscope, epithelial cells of ectopia endomembrane appear high pillar shape or low pillar shape, which located in inner cavosurface of Ves, cell-cell junction was tight, cellular nucleus was big, oval-shap, situated at basilar part, kytoplasm was abundant. However, the glandular epithelium of endomembrane became thin remarkably, cells were low pillar even applan appearance, and arranged loosely in the SLW high, middle dose groups and gestrinone group.
2. Effect of SLW on apoptosis in ectopic endometria
After four-week medication, the samples were observed under a fluorescence microscope with a standard fluorescein filter set. The green fluorescence of fluorescein could be viewed at 520±20nm. TUNEL test showed that, compared with the normal group 0.1892±0.0075, the mean fluorescence intensity value of the model group reduced to 0.1457±0.0104, the difference is remarkable (P <0.05). The value of mean fluorescence intensity of the high, middle and low dose groups of SLW were 0.2246±0.0105, 0.2209±0.0091 and 0.1955±0.0063, compared with the model group 0.1457±0.0104 , there is siginificant difference (P <0.05).
3. Effect of SLW on the mRNA expression of Bcl-2 and Fas in ectopic endometria
Analysis of hybridization in site showed that, the average optical density value of Bcl-2 mRNA is significant diference between the model group 0.1728±0.0072 and the normal group 0.1491±0.0091 (P <0.05).
However, the value of average optical density of Fas mRNA is not significant diference between the model group 0.0883±0.0038 and the normal group 0.0912±0.0016 (P >0.05). Compared with the model group, the high, middle and low dose groups could reduce the expression of Bcl-2 mRNA and promote the expression of Fas mRNA in heterotopic tissue after 4 weeks treatment. The average optical density value of Bcl-2 mRNA remarkably reduced from 0.1728±0.0072 to 0.1055±0.0040、0.1102±0.0050、0.1370±0.0070 (P <0.05). The optical density value of Fas mRNA remarkably promoted from 0.0883±0.0038 to 0.1206±0.0056、0.1164±0.0054、0.0966±0.0044 (P <0.05).
CONCLUSION
1. Traditional Chinese medicine compounds Sanleng pellet (SLW) can inhibit the growth of ectopic endometrial tissues, reduce evidencedly the volume of ectopic implants and promote apoptosis in ectopic endometria of rats.
2. SLW can induce apoptosis in ectopic endometria of rats, and its mechanism might be closely associated with inhibiting the expression of Bcl-2 mRNA and promotion the expression of Fas mRNA in the heterotopic tissue.
目的观察加味三棱丸主要成分对子宫内膜异位症大鼠异位内膜的发展及其细胞凋亡的影响;观察加味三棱丸主要成分对抗凋亡基因Bcl-2 mRNA和诱导细胞凋亡的基因Fas mRNA的作用。探讨加味三棱丸主要成分治疗子宫内膜异位症的疗效及诱导异位内膜细胞凋亡的机制。
方法:
1、动物模型的制作
选取健康雌性Wistar大鼠50只,体重180～220g,每天定时观察阴道细胞涂片,选择动情周期4～5天并连续有2个动情周期的雌性大鼠,于动情期(阴道细胞涂片为角化上皮细胞时)行造模手术。以5%水合氯醛(0.7ml/100g)腹腔注射麻醉,常规消毒,在无菌条件下开腹,结扎右侧子宫两端后,从远端剪取长约1.5㎝的子宫段,剔除脂肪组织,剪成5mm×5mm的子宫组织3块。一块送病理检验(确证为子宫组织),另外两块靠近腹膜血管处,缝于腹壁,以粘膜面朝向腹膜,浆膜面朝向腹腔,关腹。每鼠腹腔注射庆大霉素0.1ml×3天。正常清洁饲养。
2、模型纳入的标准
建模后4周,观察阴道细胞学涂片,第二次剖腹观察异位子宫内膜的生长情况,并用游标卡尺测量移植灶大小。如果移植灶体积增大,体积≥8mm3,呈隆起透亮的结节状、囊状,被结缔组织或大网膜覆盖并有血管形成,内有淡黄色透明囊液积聚。切取移植灶送检,证实移植灶中有子宫内膜上皮细胞、腺体及间质的生长。
3、试验分组与用药
除正常对照组外,另将造模成功的大鼠随机分为SLW 0.11mg.kg-1.d-1组、0.34mg.kg-1.d-1组、1.02mg.kg-1.d-1组、0.5mg.kg-1.d-1孕三烯酮组、模型对照组,灌胃给药,每天1次,连续4周。
4、检测指标
4周后结束治疗,观察阴道细胞学涂片后处死大鼠,第三次剖腹观察异位内膜生长情况,并测量移植灶体积。切取在位子宫与异位子宫内膜, HE染色进行形态学病理检察。TUNEL法检测异位内膜细胞凋亡,原位杂交检测凋亡调控基因Bcl-2和Fas mRNA的表达。
结果:
1、SLW对大鼠异位内膜组织生长的影响
用药4周后,模型对照组移植灶体积比4周前略大,呈隆起透亮的小泡状,表面血管清晰可见,内部充满积液。而孕三烯酮组和SLW 0.11mg.kg-1.d-1组、0.34mg.kg-1.d-1组、1.02mg.kg-1.d-1组移植灶体积明显缩小,囊泡表面颜色黯淡,个别移植灶颜色泛黄,呈现萎缩状。
2、SLW对大鼠异位内膜组织显微结构的影响
HE染色,光镜下观察。移植的子宫内膜组织成活,形成的囊壁由内向外分别为上皮细胞层、间质细胞层及少量纤维结缔组织,囊内为少量漂浮的白细胞和黏液。模型对照组异位内膜腺上皮细胞呈高柱状或矮柱状,排列在囊泡内腔面,细胞间连接较紧密,细胞核大,呈椭圆形,位于基底部,胞浆丰富。内膜间质也丰富,其中有个别部位上皮层内陷形成假腺体。SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组和孕三烯酮组,内膜腺上皮层明显变薄,细胞呈矮柱状甚至扁平状,松散排列,核不规则,位置不定,胞浆嗜碱性增加。内膜间质也明显减少,腺体少甚至缺失。
3、SLW对大鼠异位内膜组织体积的影响
用药前,各组大鼠异位内膜病灶体积经统计学处理无显著性差异(P >0.05)。用药4周后,孕三烯酮组和SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组、0.11mg.kg-1.d-1组异位内膜体积分别为17.01mm3、18.42mm3、25.90mm3、58.34mm3,体积明显缩小,与模型对照组102.17mm3相比均有显著性差异(P < 0.05)。而模型对照组异位内膜体积比4周前略有增加,但无显著性差异(P >0.05)。结果显示:加味三棱丸可显著缩小异位内膜体积。
4、SLW对大鼠异位内膜细胞凋亡的影响
用药4周后,异位内膜组织切片在荧光显微镜下,以520±20nm波长观察,可见绿色荧光所标记的凋亡细胞。共聚焦激光扫描,显示凋亡细胞核裂解,凋亡小体(凋亡细胞特征性的形态学改变)形成。模型对照组平均荧光强度值0.1457比正常对照组0.1892显著降低(P <0.05),模型对照组异位内膜细胞凋亡减少,生长失衡。而孕三烯酮组和SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组、0.11mg.kg-1.d-1组平均荧光强度值分别为0.2315、0.2246、0.2209、0.1955,与模型对照组0.1457相比,其平均荧光强度值均显著增高(P <0.05)。结果显示:加味三棱丸能促进异位内膜细胞凋亡。
5、SLW对大鼠异位内膜组织Bcl-2 mRNA表达的影响
用药4周后,原位杂交结果显示,抗凋亡基因Bcl-2 mRNA阳性表达的细胞染色为棕黄色,主要分布在腺上皮细胞和间质细胞的胞浆、胞膜和核膜。模型对照组异位内膜组织Bcl-2 mRNA呈阳性表达,且多数为强阳性表达,其平均光密度值0.1728与正常对照组0.1491相比有显著性差异(P <0.05)。孕三烯酮组和SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组、0.11mg.kg-1.d-1组异位内膜组织Bcl-2 mRNA的表达较模型对照组显著减少,其平均光密度值分别为0.1014、0.1055、0.1102、0.1370,与模型对照组0.1728比较均有显著性差异(P <0.05)。结果显示:加味三棱丸能降低异位内膜组织抗凋亡基因Bcl-2 mRNA的表达,从而促进异位内膜细胞凋亡。
6、SLW对大鼠异位内膜组织Fas mRNA表达的影响
用药4周后,原位杂交结果显示,凋亡基因Fas mRNA阳性表达的细胞染色为棕黄色,主要分布在内膜间质细胞的胞浆、胞膜。模型对照组异位内膜组织Fas mRNA仅有少量表达,且染色较浅,其平均光密度值0.0883与正常对照组0.0912相比无显著性差异(P >0.05)。而各治疗组异位内膜组织Fas mRNA的表达均较模型对照组有不同程度的增加,孕三烯酮组和SLW1.02mg.kg-1.d-1组、0.34mg.kg-1.d-1组、0.11mg.kg-1.d-1组平均光密度值(0.1242、0.1206、0.1164、0.0966)与模型对照组(0.0883)比较,差异有显著性(P <0.05)。结果显示:加味三棱丸能增加异位内膜组织凋亡基因Fas mRNA的表达,从而诱导异位内膜细胞凋亡。
结论:
1、加味三棱丸可抑制大鼠子宫内膜异位症的发展,缩小异位内膜体积,改善异位内膜病理学显微结构,促进异位内膜细胞凋亡。
2、加味三棱丸抑制大鼠子宫内膜异位症发展的机制,与其抑制异位内膜组织抗凋亡基因Bcl-2 mRNA表达、增加凋亡基因Fas mRNA表达,从而诱导异位内膜细胞凋亡有密切相关。
AIM
The model of endometriosis in Wistar rats by transplantation were established to investigate effects of traditional Chinese medicine compounds Sanleng pellet (SLW) on the volume of the ectopic endometrial tissue and the apoptosis of endometriotic cells. The effects of SLW on Endometriosis in Wistar rats were investigated in the expression of Bcl-2 mRNA and Fas mRNA in ectopic endometriotic tissue, meanwhile study the mechanism of SLW’s apoptosis induction in ectopic endometria.
METHOD
50 female wistar rats, 180-220 grammar, were selected to observe the smear of vaginal cell at settled time every day. Those rats whose estrus cycle is 4-5 days and which has continuous 2 estrus cycle were selected to establish rat models with endometriosis during estrus (the smear of vaginal cell is cuticular epithelial cells) according to Vernon′s method.
Four weeks after the autotransplantation, the rats were paunched the second time to investigate the growth of transplantation and mesure its volume. Those rats whose autotransplantation was not well growth were eliminated. The model rats with peritoneal implants were randomly assigned into five groups: the model control group and four treatment groups which were given respectively normal sodium, high-dose of SLW, moderate-dose of SLW, low-dose of SLW and gestrinone served as the control drugs continuously for four weeks. Meanwhile, eight normal rats were used as the blank control.
After four weeks treatment, the model rats were killed after observing the smear of vaginal cell. The rats were paunched the third time to observe the shape and mesure the volume of ectopic endometrial tissues. Eutopic endometrium and ectopic implants were cut, dipped in 4% paraform, paraffin embed, 6μm thick continuous slice made and HE stained to observe the pathologic change of eutopic and ectopic endometrium. Apoptosis in ectopic endometria was detected by TUNEL method. The score of the mRNA expression levels of Bcl-2 and Fas of ectopic implants were detected by in-situ hybridization test. Every slice was observed five fields of vision randomly under microscope. Mean fluorescence intensity and average optical density value were dealed with the image analytical software.
All data were showed by x±SD and dealed with SPSS11.5 statistical software. It is statistical significance when P<0.05.
RESULT
1. Effect of SLW on the volume size and morphological change of heterotopic tissue of EMS rats
Before treatment, the volume size value of endometrial implants has not significant diference between each treatment group and the model control group (P >0.05). After four weeks’treatment, the positive control group and SLW different doses groups could reduce the volume on the different degree, SLW high and middle dosage have the striking effect. Compared with endometriosis models, the high, middle and low dose groups could remarkably reduce heterotopic tissue volum size from 102.17±58.27mm3 to 18.42±13.21mm3, 25.90±15.26mm3and 58.34±37.05mm3 (P <0.05).
Under the light microscope, epithelial cells of ectopia endomembrane appear high pillar shape or low pillar shape, which located in inner cavosurface of Ves, cell-cell junction was tight, cellular nucleus was big, oval-shap, situated at basilar part, kytoplasm was abundant. However, the glandular epithelium of endomembrane became thin remarkably, cells were low pillar even applan appearance, and arranged loosely in the SLW high, middle dose groups and gestrinone group.
2. Effect of SLW on apoptosis in ectopic endometria
After four-week medication, the samples were observed under a fluorescence microscope with a standard fluorescein filter set. The green fluorescence of fluorescein could be viewed at 520±20nm. TUNEL test showed that, compared with the normal group 0.1892±0.0075, the mean fluorescence intensity value of the model group reduced to 0.1457±0.0104, the difference is remarkable (P <0.05). The value of mean fluorescence intensity of the high, middle and low dose groups of SLW were 0.2246±0.0105, 0.2209±0.0091 and 0.1955±0.0063, compared with the model group 0.1457±0.0104 , there is siginificant difference (P <0.05).
3. Effect of SLW on the mRNA expression of Bcl-2 and Fas in ectopic endometria
Analysis of hybridization in site showed that, the average optical density value of Bcl-2 mRNA is significant diference between the model group 0.1728±0.0072 and the normal group 0.1491±0.0091 (P <0.05).
However, the value of average optical density of Fas mRNA is not significant diference between the model group 0.0883±0.0038 and the normal group 0.0912±0.0016 (P >0.05). Compared with the model group, the high, middle and low dose groups could reduce the expression of Bcl-2 mRNA and promote the expression of Fas mRNA in heterotopic tissue after 4 weeks treatment. The average optical density value of Bcl-2 mRNA remarkably reduced from 0.1728±0.0072 to 0.1055±0.0040、0.1102±0.0050、0.1370±0.0070 (P <0.05). The optical density value of Fas mRNA remarkably promoted from 0.0883±0.0038 to 0.1206±0.0056、0.1164±0.0054、0.0966±0.0044 (P <0.05).
CONCLUSION
1. Traditional Chinese medicine compounds Sanleng pellet (SLW) can inhibit the growth of ectopic endometrial tissues, reduce evidencedly the volume of ectopic implants and promote apoptosis in ectopic endometria of rats.
2. SLW can induce apoptosis in ectopic endometria of rats, and its mechanism might be closely associated with inhibiting the expression of Bcl-2 mRNA and promotion the expression of Fas mRNA in the heterotopic tissue.
引文
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