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麻黄细辛附子汤对肾阳虚外感证模型小鼠TLRs应答及Cyt-CO介导的凋亡调控的影响
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篇名: 麻黄细辛附子汤对肾阳虚外感证模型小鼠TLRs应答及Cyt-CO介导的凋亡调控的影响
TITLE: Effects of Mahuang Xixin Fuzi Decoction on TLRs Response and Cyt-CO-mediated Apoptosis Regulation in Mice with Influenza Disease of Kidney-yang Deficiency
摘要: 目的:研究麻黄细辛附子汤对肾阳虚外感证模型小鼠Toll样受体(TLRs)应答及细胞色素C氧化酶(Cyt-CO)介导的凋亡调控的影响。方法:将48只雄性Balb/c小鼠随机分为正常组(n=12)和造模组(n=36)。造模组小鼠采用腹腔注射苯甲酸雌二醇溶液(8mg/kg)+滴鼻接种甲型H1N1流感病毒(20μL/只)的方法建立肾阳虚外感复合模型。然后将造模成功的小鼠随机分为模型组、阳性药组(磷酸奥司他韦胶囊,0.195g/kg)和麻黄细辛附子汤组(1.802g/kg,以生药总量计),每组12只。各给药组小鼠灌胃相应药液,正常组和模型组小鼠灌胃等体积生理盐水;灌胃体积均为20mL/kg,每天1次,连续给药6天。给药期间,每天测量小鼠体质量、肛温,并计算其初始体质量百分率;末次给药后,计算其脏器(脾、胸腺、肺)指数,观察其肺组织病理学变化,检测其肺组织中甲型H1N1流感病毒的病毒载量(以M基因mRNA表达水平反映)以及心脏组织中TLR3、TLR7、髓样分化因子(MyD88)、胱天蛋白酶3(Caspase-3)mRNA表达水平和Cyt-CO活性、细胞色素C(Cyt-C)含量。结果:与正常组比较,模型组小鼠的初始体质量百分率、肛温持续降低(P<0.05);脾指数、胸腺指数均显著降低(P<0.05),肺指数显著升高(P<0.05);肺组织病变严重;肺组织中病毒载量和心脏组织中TLR3、TLR7、MyD88、Caspase-3mRNA表达水平及Cyt-C含量均显著升高(P<0.05或P<0.01),心脏组织中Cyt-CO活性显著降低(P<0.01)。与模型组比较,麻黄细辛附子汤组小鼠的初始体质量百分率、肛温自给药第4天起呈上升趋势(P<0.05或P<0.01);脾指数、胸腺指数均显著升高(P<0.05),肺指数均显著降低(P<0.05);肺组织病理损伤明显改善;肺组织中病毒载量和心脏组织中TLR3、Caspase-3mRNA表达水平和Cyt-C含量均显著降低(P<0.05或P<0.01),心脏组织中Cyt-CO活性显著升高(P<0.01)。结论:麻黄细辛附子汤对肾阳虚外感病证模型小鼠具有改善作用,这可能与其可抑制TLRs应答和Cyt-CO介导的凋亡调控途径有关。
ABSTRACT: OBJECTIVE:To study the effects of Mahuang xixin fuzi decoction on Toll-like receptors (TLRs)response and cytochrome C oxidase (Cyt-CO)-mediated apoptosis regulation in mice with influenza disease of kidney-yang deficiency. METHODS:Totally 48 male Balb/c mice were randomly divided into normal group (n=12)and modeling group (n=36). The modeling group was intraperitoneally injected with estradiol benzoate solution (8 mg/kg)and intranasally injected with influenza virus H 1N1(20 μL/mice)to establish the influenza disease compound model of kidney-yang deficiency. After modeling ,the mice were randomly divided into model group ,positive drug group (Oseltamivir phosphate capsules ,0.195 g/kg),Mahuang xixin fuzi decoction group (1.802 g/kg,by crude dru g),with 12 mice in each group. Each group was given relevant medicine intragastrically,normal group and model group were given, corresponding volume of normal saline intragastrically 20 mL/kg,once a day ,for consecutive 6 days. During admi-nistration,body weight and anal temperature of mice were mail:xsy407861520@163.com measured daily ;the percentage of initial body weight was calculated. After last medication ,the organ (spleen,thymus and lung )indexes were calculated ;the pathological changes of lung tissue were observed. The viral load of influenza A virus H 1N1 in lung tissue was detected (reflected by M gene mRNA expression);mRNA expressions of TLR3,TLR7,myeloid differentiation factor (MyD88)and Caspase- 3 in cardiac tissue as well as the activity of Cyt-CO and the content of cytochrome C (Cyt-C)were also determined. RESULTS :Compared with normal group,initial body weight percentage and anal temperature of the model group continued to decrease (P<0.05);the spleen and thymus indexes were decreased significantly (P<0.05),while lung index was increased significantly (P<0.05);the lung tissue lesions were serious. Viral load in lung tissue ,mRNA expressions of TLR 3,TLR7,MyD88 and Caspase- 3 in cardiac tissue as well as the content of Cyt-C were increased significantly (P<0.05 or P<0.01),while the activity of Cyt-CO in cardiac tissue was significantly decreased (P<0.01). Compared with model group ,initial body weight percentage and anal temperature of mice in Mahuang xixin fuzi decoction group showed an increasing trend from the fourth day of administration (P<0.05 or P<0.01). The spleen and thymus indexes were increased significantly (P<0.05),while the lung index was significantly decreased (P<0.05);the pathological injury of lung tissue was significantly improved ;viral load in lung tissue ,mRNA expressions of TLR 3 and Caspase- 3 as well as the content of Cyt-C in cardiac tissue were decreased significantly (P<0.05 or P<0.01),while the activity of Cyt-CO was increased significantly in cardiac tissue (P<0.01). CONCLUSIONS :Mahuang xixin fuzi decoction can improve influenza disease of kidney-yang deficiency in mice ,the effect may related to inhibit TLRs response and apoptosis regulation pathway mediated by Cyt-CO.
期刊: 2021年第32卷第06期
编辑: 邢淑雁,范珊珊,于钦辉,马青云,孙启慧,杨勇,容蓉
AUTHORS: XING Shuyan ,FAN Shanshan ,YU Qinhui ,MA Qingyun ,SUN Qihui,YANG Yong,RONG Rong
关键字: 麻黄细辛附子汤;肾阳虚外感证;细胞色素C氧化酶;细胞色素C;凋亡;Toll样受体;小鼠
KEYWORDS: Mahuang xixin fuzi decoction ;Influenza disease of kidney-yang deficiency ;Cytochrome C oxidase ;Cytochrome
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