槲皮素对大鼠宫颈炎的治疗作用
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摘要
目的探讨槲皮素对大鼠宫颈炎的治疗作用及机制。方法将50只Sprague Dawley大鼠随机分为正常对照组、宫颈炎组、消糜阴道泡腾片组、低剂量槲皮素组和高剂量槲皮素组,每组10只。宫颈炎组、消糜阴道泡腾片组、低剂量槲皮素组和高剂量槲皮素组大鼠制备宫颈炎模型,将注射器插入大鼠阴道约1 cm处,注射250 g·L~(-1)苯酚胶浆0.1 mL,每日2次,连续5 d,至大鼠阴道有白色分泌物及阴道红肿、充血时,再次注射1次苯酚胶浆。对照组大鼠阴道内注射赋形剂0.1 mL。造模成功后,消糜阴道泡腾片组、低剂量槲皮素组和高剂量槲皮素组大鼠分别于阴道深处注射10 g·L~(-1)消糜阴道泡腾片溶液(50 mg·kg~(-1))、10 g·L~(-1)槲皮素溶液(25 mg·kg~(-1))、10 g·L~(-1)槲皮素溶液(100 mg·kg~(-1)),每日2次,连续10 d;对照组和宫颈炎组大鼠给予生理盐水。连续给药10 d后处死大鼠。采用苏木精-伊红(HE)染色观察大鼠宫颈组织病理形态学改变,酶联免疫吸附试验检测大鼠宫颈组织中细胞间黏附分子-1(ICAM-1)、白细胞介素-1β(IL-1β)和甘露糖结合凝集素(MBL)水平,Western blot法检测大鼠宫颈组织中核因子-κB(NF-κB) p65蛋白表达。结果 HE染色显示,正常对照组大鼠宫颈组织未见明显异常改变。宫颈炎组大鼠宫颈黏膜鳞状上皮细胞变性、坏死、脱落;黏膜及黏膜下固有层充血水肿,淋巴细胞和中性粒细胞等炎细胞浸润明显,可见部分脓性渗出物及坏死组织。与宫颈炎组比较,消糜阴道泡腾片组、低剂量槲皮素组和高剂量槲皮素组大鼠宫颈黏膜病理改变显著减轻。宫颈炎组大鼠宫颈组织中ICAM-1、IL-1β及MBL水平显著高于正常对照组(P<0.05),消糜泡腾片组、低剂量槲皮素组和高剂量槲皮素组大鼠ICAM-1、IL-1β及MBL水平显著低于宫颈炎组(P<0.05),消糜泡腾片组、低剂量槲皮素组和高剂量槲皮素组大鼠ICAM-1、IL-1β及MBL水平比较差异无统计学意义(P>0.05)。正常对照组、宫颈炎组、消糜泡腾片组、低剂量槲皮素组和高剂量槲皮素组大鼠宫颈组织中NF-κB p65蛋白相对表达量分别为0.13±0.02、0.68±0.03、0.39±0.02、0.37±0.02、0.38±0.02;宫颈炎组大鼠宫颈组织中NF-κB p65蛋白相对表达量显著高于正常对照组(P<0.05),消糜泡腾片组、低剂量槲皮素组和高剂量槲皮素组大鼠宫颈组织中NF-κB p65蛋白相对表达量显著低于宫颈炎组(P<0.05),低剂量槲皮素组、高剂量槲皮素组和消糜泡腾片组大鼠宫颈组织中NF-κB p65蛋白相对表达量比较差异无统计学意义(P>0.05)。结论槲皮素能够抑制宫颈组织中NF-κB p65、ICAM-1、IL-1β和MBL的表达,改善大鼠宫颈炎症反应,对大鼠宫颈炎有一定的疗效。
Objective To investigate the mechanism of quercetin in the treatment of cervicitis in rats. Methods Fifty Sprague Dawley rats were randomly divided into normal control group,cervicitis group,Xiaomi Yindao Paotengpian group,lowdose quercetin group and high-dose quercetin group,with 10 rats in each group. The rats in the cervicitis group,Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group were used to establish cervicitis model. The syringe was gently inserted into the vagina of rats about 1 cm,and 0.1 m L phenol mortar(250 g·L~(-1)) was injected,twice a day for5 days. When the vagina of rats had white secretion,vaginal swelling and congestion,the phenol mortar was injected once again. The rats in the control group were injected with 0.1 m L excipient. After successful modeling,the rats in the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group were injected with 10 g·L~(-1) Xiaomi Yindao Paotengpian solution(50 mg·kg~(-1)),10 g·L~(-1) quercetin solution(25 mg·kg~(-1)) and 10 g·L~(-1) quercetin solution(100 mg·kg~(-1)),respectively,twice a day for 10 days. The rats in the control group and cervicitis group were injected with the saline. The rats were killed after 10 days of continuous administration. The pathological changes of cervical tissues of rats were observed by hematoxylin-eosin(HE) staining. The levels of intercellular adhesion molecule-1(ICAM-1),interleukin-1β(IL-1β) and mannose-binding lectin(MBL) in cervical tissues of rats were detected by enzyme linked immunosorbent assay.The expression of nuclear factor-κB(NF-κB) p65 protein in cervical tissues of rats was detected by Western blot. Results HE staining showed that there was no abnormal change in cervical tissues of rats in the normal control group. In the cervicitis group,the changes such as cervical squamous epithelial cells degeneration,necrosis,exfoliation; congestion and edema of mucosa and submucosal lamina propria,obvious infiltration of neutrophils and lymphocytes,some purulent exudates and necrotic tissue were found. Compared with the cervicitis group,the pathological changes in cervical mucosa of rats in the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group were significantly alleviated. The levels of ICAM-1,IL-1β and MBL in cervical tissues of rats in the cervicitis group were significantly higher than those in the normal control group(P<0.05). The levels of ICAM-1,IL-1β and MBL in cervical tissues of rats in the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group were significantly lower than those in cervicitis group(P<0.05). There was no significant difference in the levels of ICAM-1,IL-1β and MBL in cervical tissues of rats among the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group(P>0.05). The relative expression of NF-κB p65 protein in cervical tissues of rats in the normal control group,cervicitis group,Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group was 0.13 ± 0.02,0.68 ± 0.03,0.39 ± 0.02,0.37 ± 0.02 and 0.38 ± 0.02,respectively. The relative expression of NF-κB p65 protein in cervical tissues of rats in the cervicitis group was significantly higher than that in the normal control group(P<0.05). The relative expression of NF-κB p65 protein in cervical tissues of rats in the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group was significantly lower than that in the cervicitis group(P<0.05). There was no significant difference in the relative expression of NF-κB p65 protein in cervical tissues of rats among the low-dose quercetin group,high-dose quercetin group and Xiaomi Yindao Paotengpian group(P>0.05). Conclusion Quercetin can inhibit the expression of NF-κB p65,ICAM-1,IL-1β and MBL in cervical tissues,improve cervical inflammatory response in rats,and has a certain effect on cervicitis in rats.
Objective To investigate the mechanism of quercetin in the treatment of cervicitis in rats. Methods Fifty Sprague Dawley rats were randomly divided into normal control group,cervicitis group,Xiaomi Yindao Paotengpian group,lowdose quercetin group and high-dose quercetin group,with 10 rats in each group. The rats in the cervicitis group,Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group were used to establish cervicitis model. The syringe was gently inserted into the vagina of rats about 1 cm,and 0.1 m L phenol mortar(250 g·L~(-1)) was injected,twice a day for5 days. When the vagina of rats had white secretion,vaginal swelling and congestion,the phenol mortar was injected once again. The rats in the control group were injected with 0.1 m L excipient. After successful modeling,the rats in the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group were injected with 10 g·L~(-1) Xiaomi Yindao Paotengpian solution(50 mg·kg~(-1)),10 g·L~(-1) quercetin solution(25 mg·kg~(-1)) and 10 g·L~(-1) quercetin solution(100 mg·kg~(-1)),respectively,twice a day for 10 days. The rats in the control group and cervicitis group were injected with the saline. The rats were killed after 10 days of continuous administration. The pathological changes of cervical tissues of rats were observed by hematoxylin-eosin(HE) staining. The levels of intercellular adhesion molecule-1(ICAM-1),interleukin-1β(IL-1β) and mannose-binding lectin(MBL) in cervical tissues of rats were detected by enzyme linked immunosorbent assay.The expression of nuclear factor-κB(NF-κB) p65 protein in cervical tissues of rats was detected by Western blot. Results HE staining showed that there was no abnormal change in cervical tissues of rats in the normal control group. In the cervicitis group,the changes such as cervical squamous epithelial cells degeneration,necrosis,exfoliation; congestion and edema of mucosa and submucosal lamina propria,obvious infiltration of neutrophils and lymphocytes,some purulent exudates and necrotic tissue were found. Compared with the cervicitis group,the pathological changes in cervical mucosa of rats in the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group were significantly alleviated. The levels of ICAM-1,IL-1β and MBL in cervical tissues of rats in the cervicitis group were significantly higher than those in the normal control group(P<0.05). The levels of ICAM-1,IL-1β and MBL in cervical tissues of rats in the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group were significantly lower than those in cervicitis group(P<0.05). There was no significant difference in the levels of ICAM-1,IL-1β and MBL in cervical tissues of rats among the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group(P>0.05). The relative expression of NF-κB p65 protein in cervical tissues of rats in the normal control group,cervicitis group,Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group was 0.13 ± 0.02,0.68 ± 0.03,0.39 ± 0.02,0.37 ± 0.02 and 0.38 ± 0.02,respectively. The relative expression of NF-κB p65 protein in cervical tissues of rats in the cervicitis group was significantly higher than that in the normal control group(P<0.05). The relative expression of NF-κB p65 protein in cervical tissues of rats in the Xiaomi Yindao Paotengpian group,low-dose quercetin group and high-dose quercetin group was significantly lower than that in the cervicitis group(P<0.05). There was no significant difference in the relative expression of NF-κB p65 protein in cervical tissues of rats among the low-dose quercetin group,high-dose quercetin group and Xiaomi Yindao Paotengpian group(P>0.05). Conclusion Quercetin can inhibit the expression of NF-κB p65,ICAM-1,IL-1β and MBL in cervical tissues,improve cervical inflammatory response in rats,and has a certain effect on cervicitis in rats.
引文
[1]孙玉兰,顾云鹏,王宏达,等.槲皮素对宫颈炎模型大鼠治疗效果的初步实验研究[J].现代生物医学进展,2018,18(2):222-225.
[2]任国平,王保莲,毕春燕,等.消糜膏对宫颈炎患者的临床疗效及对宫颈局部EGF、IL-1β、TNF-α表达的影响[J].世界中医药,2017,12(4):822-824.
[3]谢辛,苟文丽.妇产科学[M].8版.北京:人民卫生出版社,2013:255-257.
[4]BEYAZIT F,BUYUK B.An immunohistochemistry and histopathological study of ankaferd blood stopper in a rat model of cervical inflammation[J].Rev Assoc Med Bras,2019,65(2):183-190.
[5]朱迎萍,吴燕平.自拟宫颈方对宫颈炎模型大鼠宫颈组织VEGF-A、NF-κB表达的影响[J].浙江中西医结合杂志,2017,27(9):752-755.
[6]LEE I Y,IM J M,CHO H,et al.MST1 negatively regulates TNF-α-induced NF-κB signaling through modulating LUBAC activity[J].Mol Cell,2019,73(6):1138-1149.
[7]韩玉梅,谢晶莹,邓盈盈,等.ICAM-1在NF-κB通路中表达的研究进展[J].西北民族大学学报(自然科学版),2018,39(3):40-45.
[8]赵瑞萍,陈卫东,常保超,等.雷公藤多苷抑制糖尿病大鼠肾组织MBL及MASP2的表达并减轻肾损伤[J].细胞与分子免疫学杂志,2017,33(11):1498-1503.
[9]张昕,魏江存,阙祖亮,等.中药抗炎作用研究概况[J].中国民族民间医药,2017,26(21):67-69.
[10]李聪,胡强,张燕翔,等.槲皮素的药理学活性研究进展[J].湖北中医杂志,2018,40(6):63-66.
[11]许银凤,叶云,孙琴,等.槲皮素对LPS刺激的小胶质细胞炎症因子的下调作用[J].基因组学与应用生物学,2017,36(3):1173-1179.
[12]验研究,王香青,马振军,等.槲皮素通过抑制β-catenin入核抑制人宫颈癌细胞增殖、迁移的实验研究[J].天津中医药,2019,36(2):195-199.
[2]任国平,王保莲,毕春燕,等.消糜膏对宫颈炎患者的临床疗效及对宫颈局部EGF、IL-1β、TNF-α表达的影响[J].世界中医药,2017,12(4):822-824.
[3]谢辛,苟文丽.妇产科学[M].8版.北京:人民卫生出版社,2013:255-257.
[4]BEYAZIT F,BUYUK B.An immunohistochemistry and histopathological study of ankaferd blood stopper in a rat model of cervical inflammation[J].Rev Assoc Med Bras,2019,65(2):183-190.
[5]朱迎萍,吴燕平.自拟宫颈方对宫颈炎模型大鼠宫颈组织VEGF-A、NF-κB表达的影响[J].浙江中西医结合杂志,2017,27(9):752-755.
[6]LEE I Y,IM J M,CHO H,et al.MST1 negatively regulates TNF-α-induced NF-κB signaling through modulating LUBAC activity[J].Mol Cell,2019,73(6):1138-1149.
[7]韩玉梅,谢晶莹,邓盈盈,等.ICAM-1在NF-κB通路中表达的研究进展[J].西北民族大学学报(自然科学版),2018,39(3):40-45.
[8]赵瑞萍,陈卫东,常保超,等.雷公藤多苷抑制糖尿病大鼠肾组织MBL及MASP2的表达并减轻肾损伤[J].细胞与分子免疫学杂志,2017,33(11):1498-1503.
[9]张昕,魏江存,阙祖亮,等.中药抗炎作用研究概况[J].中国民族民间医药,2017,26(21):67-69.
[10]李聪,胡强,张燕翔,等.槲皮素的药理学活性研究进展[J].湖北中医杂志,2018,40(6):63-66.
[11]许银凤,叶云,孙琴,等.槲皮素对LPS刺激的小胶质细胞炎症因子的下调作用[J].基因组学与应用生物学,2017,36(3):1173-1179.
[12]验研究,王香青,马振军,等.槲皮素通过抑制β-catenin入核抑制人宫颈癌细胞增殖、迁移的实验研究[J].天津中医药,2019,36(2):195-199.