基于sGC-cGMP-PKG信号通路研究桂郁金水提物对异丙肾上腺素诱导小鼠心肌肥厚的作用及机制  

Effects of aqueous extract of Curcuma kwangsiensis root tubers on isoproterenol-induced cardiac hypertrophy in mice based on the sGC-cGMP-PKG signaling pathway

作　　者：何永芸 黎红 胡明霞 姚玥 莫晓丹 杨秀芬 HE Yongyun;LI Hong;HU Mingxia;YAO Yue;MO Xiaodan;YANG Xiufen(School of Pharmacy,Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi KeyLaboratory of Zhuang and Yao Ethnic Medicine,Nanning 530200,China;Key Laboratory of TraditionalChinese Medicine Neuro-Metabolism and Immunopharmacology of Guangxi Education Department,Nanning 530200,China)

机构地区：[1]广西中医药大学药学院,南宁530200 [2]广西壮瑶药重点实验室,南宁530200 [3]广西高校中药神经-代谢及免疫药理重点实验室,南宁530200

出　　处：《中国实验动物学报》2024年第10期1260-1269,共10页Acta Laboratorium Animalis Scientia Sinica

基　　金：广西自然科学基金(2020GXNSFAA297165);广西壮瑶药重点实验室建设项目(GXZYZZ2022-08);中药学广西一流学科(桂教科研[2022]1号)。

摘　　要：目的基于sGC-cGMP-PKG信号通路研究桂郁金(Curcuma kwangsiensis root tubers,GYJ)水提物对异丙肾上腺素(isoproterenol,ISO)诱导小鼠心肌肥厚的改善作用及相关机制。方法将72只KM雄性小鼠随机分为正常对照组、模型组、普萘洛尔(40 mg/kg)组以及GYJS低(1 g/kg)、中(2 g/kg)、高(4 g/kg)剂量组。除正常对照组外,其余各组小鼠在第1~3天皮下注射ISO(10 mg/kg),第4~14天皮下注射ISO(5 mg/kg),皮下注射ISO 4 h后,各组小鼠灌胃相应的药物,给药周期为14 d。取材后,称量小鼠全心重和左心室重量;苏木素-伊红(HE)染色和马松(Masson)染色观察小鼠心肌组织的病理情况;免疫组织化学(immunohistochemical,IHC)染色观察鸟苷酸环化酶1β3(guanylate cyclase 1,soluble,beta 3,GUCY1B3)、转化生长因子-β1(ransforming growth factor beta 1,TGF-β1)心肌组织中的表达情况;试剂盒检测小鼠血清中乳酸脱氢酶(lactate dehydrogenase,LDH)、肌酸激酶(creatine kinase,CK)活性和一氧化氮(nitric oxide,NO)含量,检测心肌组织中超氧化物歧化酶活性(superoxide dismutase,SOD)和丙二醛(malondialdehyde,MDA)含量;ELISA法测定小鼠血清中环磷酸鸟苷(cyclic guanosine monophosphate,cGMP)含量;逆转录荧光定量PCR(quantitative reverse transcription PCR,RT-qPCR)检测心房钠尿肽(atrial natriuretic peptide,ANP)、脑钠肽(brain natriuretic peptide,BNP)、GUCY1B3、环磷酸鸟苷依赖蛋白激酶1(cGMP-dependent protein kinase 1,PKGⅠ)、磷酸二酯酶5A(phosphodiesterase 5A,PDE5A)mRNA表达水平。结果与模型组比较,普萘洛尔组和GYJS各剂量组可以显著降低小鼠全心重指数和左心室重量指数(P<0.001或P<0.0001),明显改善小鼠心肌组织肥厚和心肌纤维化,显著升高GUCY1B3在小鼠心肌组织中表达(P<0.05或P<0.01),显著降低TGF-β1的表达(P<0.05或P<0.01),心肌损伤标志物LDH、CK活力显著降低(P<0.05或P<0.01),NO、cGMP含量显著升高(P<0.05或P<0.01),心肌氧化应激指标MDA含量显著降低(P<0.Objective To investigate the role and related mechanism of the soluble guanylate cyclase(sGC)cGMP-protein kinase G(PKG)signaling pathway in the amelioration of isoproterenol(ISO)-induced cardiac hypertrophy in mice by aqueous extract of Curcuma kwangsiensis root tubers(GYJS).Methods 72 KM male mice were divided randomly into 6 groups:normal,model,propranolol control(40 mg/kg),and GYJS low-(1 g/kg),medium-(2 g/kg),and high-dose(4 g/kg)groups.Mice in the experimental groups were injected subcutaneously with ISO 10 mg/kg on days 1~3 and ISO 5 mg/kg on days 4~14 to establish a mouse cardiac hypertrophy model.4h after each subcutaneous injection of ISO,the mice in each group were administered the corresponding drugs orally for a dosing cycle of 14 days.The hearts were then removed and whole heart and left ventricle weight were measured.Myocardial tissue pathology was observed using hematoxylin and eosin and Masson staining,and sGC subunit beta-1(GUCY1B3)and transforming growth factor(TGFβ1)were detected by immunohistochemistry.Serum lactate dehydrogenase(LDH),creatine kinase(CK),and Nitric Oxide(NO),and myocardial superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were measured using respective kits.Serum cGMP was detected by enzyme-linked immunosorbent assay and quantitative reverse transcription PCR(RT-qPCR),and atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),GUCY1B3,PKGⅠ,and phosphodiesterase(PDE)5A mRNA expression levels were also determined by RT-qPCR.Results Compared with the model group,whole heart and left ventricle weights were significantly reduced in mice treated with propranolol or GYJS(P<0.001 or P<0.0001)and myocardial hypertrophy and myocardial fibrosis were significantly reversed.All the treatments significantly elevated myocardial expression of GUCY1B3(P<0.05 or P<0.01)and significantly reduced expression of TGF-β1(P<0.05 or P<0.01).The myocardial damage markers LDH and CK were significantly reduced(P<0.05 or P<0.01)while NO and cGMP were significantly elevated(P<0.05

关 键 词：桂郁金 小鼠 心肌肥厚 sGC-cGMP-PKG信号通路 机制 

分 类 号：Q95-33[生物学—动物学]