摘要
目的探讨5-羟色胺(5-HT)2B受体拮抗剂通过兔抗大鼠细胞外调蛋白激酶1/2(ERK1/2)通路干预大鼠垂体瘤纤维化的作用机制。方法将60只大鼠分为对照组、模型组、5-HT2B受体拮抗剂组及5-HT2B受体激动剂组,各15只大鼠,除对照组外,其余各组以雌激素建立大鼠垂体瘤模型。分别用2 mg/kg SB204741、2 mg/kg BW723C86对5-HT2B受体拮抗剂组及5-HT2B受体激动剂组大鼠进行腹腔注射,对照组及模型组大鼠分别注射等量生理盐水,共14 d。干预结束后,测量各组大鼠垂体肿瘤最大直径并计算肿瘤体积;苏木精-伊红(HE)染色观察垂体瘤组织病理变化;Masson染色观察垂体瘤组织胶原分布情况;逆转录-聚合酶链式反应(RT-PCR)检测兔抗大鼠转化生长因子-β_(1)(TGF-β_(1))、结缔组织生长因子(CTGF)、ERK1/2 mRNA相对表达情况;蛋白免疫印迹法(Western Blot)检测TGF-β_(1)、CTGF、ERK1/2、磷酸化ERK1/2(p-ERK1/2)蛋白相对表达情况。结果与模型组比较,5-HT2B受体拮抗剂组大鼠垂体瘤最大直径、体积及垂体质量均降低,5-HT2B受体激动剂组大鼠垂体瘤最大直径、体积及垂体质量均升高,差异均有统计学意义(P<0.05)。HE染色显示:对照组垂体细胞数目较多,排列整齐有序;模型组大鼠垂体细胞有瘤样改变,细胞数量明显增多,胞核大小不一,细胞间质增生明显,有炎性细胞浸润;5-HT2B受体拮抗剂组有瘤样改变,细胞数量增多较少,偶有间质增生现象,炎性反应较轻;5-HT2B受体激动剂组垂体细胞数量大量增多,核大且核仁明显,细胞间质增生现象严重,大量炎性细胞浸润。Masson染色显示:对照组垂体细胞多且分布均匀,无胶原产生;模型组有不连续胶原纤维,分布密集;5-HT2B受体拮抗剂组胶原分布较少,无连续胶原纤维;5-HT2B受体激动剂组胶原纤维较多且连续。与对照组比较,模型组、5-HT2B受体拮抗剂组及5-HT2B受体激动剂组垂体组织中TGF-β_(1)、CTGF mRNA相对表达量及TGF-β_(1)、CTGF、p-ERK1/2蛋白相对表达量均升高(P<0.05);与模型组比较,5-HT2B受体拮抗剂组垂体组织中TGF-β_(1)、CTGF mRNA相对表达量及TGF-β_(1)、CTGF、p-ERK1/2蛋白相对表达量均降低,5-HT2B受体激动剂组均升高(P<0.05);5-HT2B受体激动剂组TGF-β_(1)、CTGF mRNA相对表达量及TGF-β_(1)、CTGF、p-ERK1/2蛋白相对表达量均高于5-HT2B受体拮抗剂组(P<0.05);对照组、模型组、5-HT2B受体拮抗剂组及5-HT2B受体激动剂组ERK1/2 mRNA及ERK1/2蛋白相对表达量比较,差异均无统计学意义(P>0.05)。结论5-HT2B受体拮抗剂对大鼠垂体瘤纤维化具有改善作用,可能是通过抑制ERK通路发挥作用。
Objective To investigate the effect of 5-hydroxytryptamine(5-HT)2B receptor antagonist on rat pituitary tumor fibrosis through extracellular protein kinase 1/2(ERK1/2)pathway.Methods Sixty rats were divided into control group,model group,5-HT2B receptor antagonist group,and 5-HT2B receptor agonist group,with 15 rats in each group.Except for the control group,the rats in other groups were used to pituitary tumor model with estrogen.The rats in 5-HT2B receptor antagonist group and 5-HT2B receptor agonist group were intraperitoneally injected with 2 mg/kg SB204741 and 2 mg/kg BW723C86,respectively.The rats in control group and model group were injected intraperitoneally with the same volume of normal saline,for 14 days.After the intervention,the maximum diameter of the pituitary tumors in each group was measured and the tumor volume was calculated.The pathological changes of pituitary tumors was observed using hematoxylin-eosin(HE)staining.The collagen distribution of pituitary tumors were observed using Masson staining.The mRNA relative expressions of transforming growth factor-β_(1)(TGF-β_(1)),connective tissue growth factor(CTGF),and ERK1/2 were detected by reverse transcription-polymerase chain reaction(RT-PCR).The relative expressions of TGF-β_(1),CTGF,ERK1/2,and p-ERK1/2 proteins were detected by Western Blot.Results Compared with model group,the maximum diameter,volume,and pituitary mass of rat pituitary tumors in 5-HT2B receptor antagonist group decreased,while those in 5-HT2B receptor agonist group increased(P<0.05).HE staining showed that the number of pituitary cells in control group was large,and they were neatly arranged.There were tumor-like changes in pituitary cells in model group,the number of cells was increased significantly,the size of nuclei was different,the interstitial proliferation was obvious,and there was inflammatory cell infiltration.There were tumor-like changes in 5-HT2B receptor antagonist group,the number of cells increased less,there was occasional interstitial hyperplasia,and the inflammatory reaction was lighter.The number of pituitary cells in 5-HT2B receptor agonist group increased greatly,with large nuclei and obvious nucleoli,intercellular hyperplasia was serious,and a large number of inflammatory cells infiltrate.Masson staining showed that pituitary cells were abundant and evenly distributed without collagen production in control group.There were discontinuous collagen fibers with dense distribution in model group.There was less collagen distribution and no continuous collagen fiber in 5-HT2B receptor antagonist group.There were more and continuous collagen fibers in 5-HT2B receptor agonist group.Compared with control group,the mRNA relative expression levels of TGF-β_(1) and CTGF and protein relative expression levels of TGF-β_(1),CTGF,and P-ERK1/2 in pituitary tissues of model group,5-HT2B receptor antagonist group,and 5-HT2B receptor agonist group were increased.Compared with model group,the relative expressions levels of TGF-β_(1),CTGF mRNA,and TGF-β_(1),CTGF,p-ERK1/2 proteins in the pituitary tissue of 5-HT2B receptor antagonist group were decreased,and those in 5-HT2B receptor agonist group were increased,and those in 5-HT2B receptor agonist group were higher than those in 5-HT2B receptor antagonist group(P<0.05).The relative expression levels of ERK1/2 mRNA and ERK1/2 protein in control group,model group,5-HT2B receptor antagonist group,and 5-HT2B receptor agonist group were not significantly different(P>0.05).Conclusion 5-HT2B receptor antagonist can improve rat pituitary tumor fibrosis,possibly by inhibiting ERK pathway.
作者
王明国
饶克成
乔卿均
高飞
WANG Mingguo;RAO Kecheng;QIAO Qingjun;GAO Fei(Nanyang Second People′s Hospital,Nanyang 473000,Henan,China)
出处
《中西医结合心脑血管病杂志》
2022年第9期1588-1593,共6页
Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease
基金
河南省医学科技攻关计划(No.LHGJ20191470)。
关键词
垂体瘤
纤维化
5-羟色胺2B受体拮抗剂
细胞外调蛋白激酶
大鼠
实验研究
pituitary tumor
fibrosis
5-hydroxytryptamine 2B receptor antagonist
extracellular regulated protein kinase
rats
experiment research
