苍附导痰汤对多囊卵巢综合征大鼠oatp4a1、代谢紊乱与肠道菌群的影响

Effects of Cangfu Daotan Decoction on OATP4A1, metabolic disorders and intestinal flora in rats with polycystic ovary syndrome

关键词:  苍附导痰汤  多囊卵巢综合征  oatp4a1  代谢紊乱  肠道菌群

Key words:  Cangfu Daotan Decoction  polycystic ovary syndrome  oatp4a1  metabolic disorders  intestinal flora

摘要:

[目的] 探究苍附导痰汤对多囊卵巢综合征大鼠oatp4a1、代谢紊乱与肠道菌群的影响。[方法] 选取40只SPF级SD雌性大鼠，随机分为正常（N）组、模型（M）组、低剂量苍附导痰汤（D）组、高剂量苍附导痰汤（H）组，每组10只，对M组、D组、H组采用脱氢表雄酮（DHEA）及高脂饲料建立多囊卵巢综合征模型，N组不建立该模型，建模后对D组灌胃1.42 g/kg苍附导痰汤，对H组灌胃5.68 g/kg苍附导痰汤，N组、M组同期给予同体积生理盐水灌胃，苏木精-伊红（HE）染色法检测卵巢组织病理形态，免疫组化法检测卵巢组织中有机阴离子转运多肽4a1（oatp4a1）蛋白表达，血糖仪、酶联免疫吸附测定（ELISA）法及放射免疫法检测大鼠血清中代谢相关水平，肠道菌群选择性培养基检测大鼠肠道菌群数量变化。[结果] 与N组比较，M组大鼠动情前期、卵巢指数明显升高（P<0.05），动情期明显降低（P<0.05）；与M组比较，D组、H组大鼠动情前期、卵巢指数显著降低（P<0.05），动情期显著升高（P<0.05），且H组比D组变化明显（P<0.05）。N组大鼠卵巢组织及卵泡生长正常，可见多个黄体和不同发育时期卵泡，且间质正常，间质中可见深染的肥大细胞，M组大鼠可见卵巢皮质呈多囊样改变，黄体数量显著减少，可见较多的闭锁卵泡，卵泡细胞多为坏死、增厚，D组、H组大鼠卵巢病理显著改善，黄体数量逐渐增多。与N组比较，M组大鼠卵巢组织oatp4a1蛋白表达显著降低（P<0.05）；与M组比较，D组、H组大鼠卵巢组织oatp4a1蛋白表达明显升高（P<0.05），且H组比D组显著升高（P<0.05）。与N组比较，M组血清中空腹血糖（FBG）、空腹胰岛素（FINS）、胰岛素抵抗指数（HOMA-IR）显著升高（P<0.05），脂联素（APN）显著降低（P<0.05）；与M组比较，D组、H组血清中FBG、FINS、HOMA-IR显著降低（P<0.05），APN显著升高（P<0.05），且H组比D组变化显著（P<0.05）。与N组比较，M组大鼠肠杆菌数量显著升高（P<0.05），肠球菌、乳酸杆菌、双歧杆菌数量明显降低（P<0.05）；与M组比较，D组、H组大鼠肠杆菌数量显著降低（P<0.05），肠球菌、乳酸杆菌、双歧杆菌数量明显升高（P<0.05），且H组比D组变化显著（P<0.05）。[结论] 苍附导痰汤可显著促进多囊卵巢综合征大鼠oatp4a1蛋白表达，改善大鼠代谢紊乱和肠道菌群。

Abstract:

[Objective] To explore the effects of Cangfu Daotan Decoction on oatp4a1, metabolic disorders and intestinal flora in rats with polycystic ovary syndrome. [Methods] The 40 SPF SD female rats were selected and randomly divided into normal group(N), model group(M), low dose pale attached guide phlegm decoction(D) group, high dose pale attached guide phlegm decoction(H) group, each group of 10, only the M group, D and H group adopts the use of DHEA(DHEA) and high fat feed polycystic ovary syndrome model was established. The N group was not set up the model. After modeling, D group was given 1.42 g/kg Cangfu Daotan Decoction;H group was given 5.68 g/kg Cangfu Daotan Decoction;N group and M group were given the same volume of normal saline;HE staining method was used to detect the pathological morphology of ovarian tissue, immunohistochemistry method was used to detect oatp4a1 protein expression in ovarian tissue. Serum metabolism-related levels of rats were detected by glucose analyzer, ELISA and radioimmunoassay, and the number of intestinal flora was detected by selective medium. [Results] Compared with N group, M group rats pro oestrus and ovarian index increased significantly(P<0.05), estrus significantly decreased(P<0.05), compared with M group, the D and H group rats pro oestrus and ovarian index decreased significantly(P<0.05), a significant rise in estrus(P<0.05), and than changes in D group and H group was obviously(P<0.05). N group of normal ovarian tissue of rats and follicular growth, visible multiple corpus luteum follicle and different development period, and interstitial, shape good, normal stroma in deep dyeing of mast cells, M group visible ovarian cortex of rats with polycystic change, corpus luteum number decreased significantly, visible more atresia follicles, follicular cell necrosis, thickening, more visible and obvious inflammatory cell infiltration. The pathological changes of ovary in D group and group H group were significantly improved, and the number of luteum increased gradually. Compared with N group, oatp4a1 protein expression in ovary tissues of rats in M group was significantly decreased(P<0.05), oatp4a1 protein expression in ovary tissues of rats in D group and H group was significantly increased compared with M group(P<0.05), and H group was significantly higher than D group(P<0.05). Compared with N group, serum FBG, FINS and HOMA-IR in M group were significantly increased(P<0.05), and SERUM APN in M group was significantly decreased(P<0.05). Compared with M group, serum FBG, FINS and HOMA-IR in D group and H group were significantly decreased(P<0.05), and serum APN was significantly increased(P<0.05). The change of H group was significantly higher than that of D group(P<0.05). Compared with N group, M group enterobacter number of rats significantly increased(P<0.05), enterococcus and lactobacillus, bifidobacterium quantity decreased obviously(P<0.05). Compared with M group, D and H group in the rat intestinal bacteria number decreased significantly(P<0.05), enterococcus and lactobacillus, bifidobacterium quantity increased significantly(P<0.05), and the change of H group was significantly higher than that of D group(P<0.05). [Conclusion] Cangfu Daotan Decoction can significantly promote oatp4a1 protein expression in rats with polycystic ovary syndrome, improve metabolic disorders and intestinal flora of rats.