摘要
目的:探究白花蛇舌草提取物调控TGF-β/Smad信号转导通路影响肾癌细胞增殖、侵袭及肾癌小鼠Th1/Th2细胞因子表达的机制。方法:以小鼠肾癌Renca细胞为研究对象,用白花蛇舌草提取物干预后,在细胞水平采用划痕实验检测其对小鼠肾癌细胞迁移能力的影响;CCK-8实验观察白花蛇舌草提取物对小鼠肾癌细胞增殖能力的作用。采用右侧腋下皮下处接种方法建立小鼠肾癌模型,研究白花蛇舌草提取物在小鼠体内对肾细胞癌增殖和侵袭作用的影响。HE染色观测小鼠移植瘤块组织形态;Western blot检测小鼠移植瘤组织TGF-β/Smad通路蛋白TGF-β、Smad2和p-Smad2表达;ELISA检测小鼠血清中IL-2、IL-10水平,研究白花蛇舌草提取物对Th1/Th2肾癌免疫偏移是否有抑制作用。结果:CCK-8实验结果表明:白花蛇舌草提取物抑制了小鼠肾癌Renca细胞的增殖能力(P<0.05);划痕实验结果表明:白花蛇舌草提取物有效抑制了小鼠肾癌Renca细胞的愈合能力(P<0.05);通过构建小鼠肾癌模型发现白花蛇舌草提取物可抑制小鼠移植瘤体积的增长(P<0.05);HE染色显示:模型组瘤细胞致密、核大分裂相多、坏死少,白花蛇舌草组和通路抑制剂组癌细胞坏死增多,核固缩或溶解;Western blot结果表明:与模型组相比,白花蛇舌草提取物组和通路抑制剂组TGF-β、p-Smad2表达降低(P<0.001);ELISA结果显示:与模型组比较,白花蛇舌草提取物组和通路抑制剂组血清IL-2表达升高(P<0.0001)。白花蛇舌草提取物组IL-10表达降低(P<0.05),通路抑制剂组IL-10表达降低(P>0.05)。结论:白花蛇舌草提取物可能通过抑制TGF-β/Smads信号通路蛋白表达,进一步调控Th1/Th2细胞因子的分泌,从而抑制小鼠肾癌细胞体内外的增殖和迁移。
Objective:To explore the mechanism by which the extract of Hedyotis diffusa Willd regulates TGF-β/Smad signaling transduction pathway and affects proliferation,invasion of renal cancer cells and expressions of Th1/Th2 cytokines in mice with renal cancer.Methods:Using mouse renal cancer Renca cells as the research object,after intervention with extracts of Hedyotis diffusa Willd,impact on migration ability of mouse renal cancer cells was detected at the cellular level using scratch assay;observing effect of CCK-8 experiment on proliferation ability of mouse renal cancer cells.A mouse kidney cancer model was established by subcutaneous inoculation under the right armpit,and effect of extract of Hedyotis diffusa Willd on proliferation and invasion of renal carcinoma in mice was studied.HE staining was used to observe histomorphology of transplanted tumor mass in mice;Western blot was used to detected expressions of TGF-β/Smad pathway protein TGF-β,Smad2 and p-Smad2 in mouse transplanted tumor tissue;levels of IL-2 and IL-10 in mouse serum were detected by ELISA to investigate whether the extract of Hedyotis diffusa Willd has an inhibitory effect on Th1/Th2 renal carcinoma immune shift.Results:CCK-8 experiment showed that the extract of Hedyotis diffusa Willd inhibited proliferation ability of mouse renal cancer Renca cells(P<0.05);scratch test results showed that extract of Hedyotis diffusa Willd effectively inhibited the healing ability of mouse renal cancer Renca cells(P<0.05);by constructing a mouse kidney cancer model,it was found that extract of Hedyotis diffusa Willd could inhibit the growth of transplanted tumor volume in mice(P<0.05);HE staining revealed that tumor cells in model group were dense,with numerous mitotic figures and minimal necrosis;whereas in Hedyotis diffusa Willd group and pathway inhibitor group,there was increased necrosis of cancer cells,accompanied by karyopyknosis or lysis;Western blot showed that compared with model group,expression of TGF-β and p-Smad2 in extract of Hedyotis diffusa Willd group and pathway inhibitor group were decreased(P<0.001);ELISA results showed that compared with model group,serum IL-2 expression increased in both Hedyotis diffusa Willd group and pathway inhibitor group(P<0.0001).IL-10 expression decreased in Hedyotis diffusa Willd group(P<0.05)and pathway inhibitor group(P>0.05).Conclusion:Extract of Hedyotis diffusa Willd may inhibit protein expressions in TGF-β/Smads signaling pathway,further regulate the secretion of Th1/Th2 cytokines,thereby inhibiting proliferation and migration of mouse renal cancer cells both in vitro and in vivo.
作者
王兵
尚官威
宋圣佑
张林超
WANG Bing;SHANG Guanwei;SONG Shengyou;ZHANG Linchao(The Second Clinical Medical College of Henan University of TCM,Henan Province Hospital of TCM,Zhengzhou 450002,China)
出处
《中国免疫学杂志》
北大核心
2025年第9期2185-2191,共7页
Chinese Journal of Immunology
基金
河南省中医药科学研究专项课题(2021JDZX2043)。
