CN114533786A - 一种曲札茋苷的新用途 - Google Patents
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Abstract
本发明公开了拉萨大黄提取物,特别是曲札茋苷在制备ACE抑制活性相关药物中的应用。基于对SARS‑CoV‑2感染导致心肾损伤的机制分析,曲札茋苷具有作为通过调节RAS系统以及靶向作用于ACE潜在的药物,有助于减缓COVID‑19患者的心肾损伤,在制备防治新型冠状病毒肺炎导致的心肾等多器官损伤药物中的应用。该类药物的活性成份可以从植物中提取,产率高,可用于工业化生产,所用药物容易获得,市场前景好。
Description
技术领域
本发明属于医药技术领域,涉及拉萨大黄提取物在心肾等多器官损伤药物中的新用途,特别涉及曲札茋苷在制备防治新型冠状病毒肺炎导致的心肾等多器官损伤药物中的新用途。
背景技术
目前,血管紧张素抑制剂(ACEI)在临床上得到广泛应用,机制是通过肾素-血管紧张素-醛固酮系统(RAAS)对调节体内神经、内分泌及血压等方面起着重要作用。ACEI具有减少尿蛋白和改善肾功能的作用,在慢性充血性心力衰竭、心肌梗死、糖尿病、肝损伤、及DN、消化系统疾病、神经系统疾病等方面ACEI也得到广泛应用。
新型冠状病毒肺炎(Coronavirus disease 2019,COVID-19)是新型冠状病毒(SARS-CoV-2)感染导致的,与SARS病毒(SARS-CoV)、 MERS病毒(RS-CoV)同属β属冠状病毒,是一种具有传染性质的急性呼吸道疾病。当前,新型冠状病毒仍在全球范围内肆虐,COVID-19 患者除表现为典型肺炎症状之外,也可出现心肾损伤,若合并心肾系统等基础疾病,则有较高的风险发展为重症甚至死亡。
大量的研究表明,新冠病毒入侵细胞的机制是:SARS-CoV-2通过其突刺蛋白与受体血管紧张素转换酶(Angiotensin converting enzyme,ACE)2相互作用,在TMPRSS2的存在下进入宿主细胞(中国药理学与毒理学杂志2020年7月第34卷第7期,血管紧张素转换酶2在新型冠状病毒肺炎(COVID-19)发病机制及治疗中的作用研究进展)。新冠病毒主要是利用ACE2进入细胞来进行破坏和复制病毒, ACE2在肾脏、心脏、小肠、十二指肠、胆囊、肾、睾丸、及直肠等组织高度分布,提示其在心肾功能中的重要作用,血管、肾脏、肝管、胰腺、肠道和呼吸道内侧的细胞也都布满了ACE2受体,病毒可以利用这些受体来抓取和感染细胞,因此新冠肺炎不仅仅只是对肺部造成损害,对身体的其他系统也造成了严重的损伤作用,使身体出现多种疾病表现,所有表达ACE2的组织器官都可能是新型冠状病毒与免疫细胞的战场,最终导致多器官衰竭,危及生命。郭建淑等人在《病毒学报,新型冠状病毒受体血管紧张素转换酶2的研究进展》同样报导血管紧张素转换酶ACE2是新型冠状病毒的受体,并根据既往的研究, ACE2是肾素血管紧张素系统(Renin angiotensin system,RAS)的关键酶之一,与其同源物ACE在该系统中起协同调节作用,同时,其生理调节功能也广泛作用于心血管、肾脏、肝脏、肠道。病毒感染人体后出现ACE2表达水平下降,具有舒张血管、改善内皮功能、抗增殖功能的Ang(1-7)产生减少,从而产生心脏损害,因此作为协同调节作用的ACE也作为调节的靶点作用。研究证明血管紧张素转换酶抑制剂(an-giotensin converting enzymeinhibition,ACEI)和AT1R拮抗剂 (angiotensinⅡtype 1receptor blocker,ARB)能够提高血浆Ang(1-7) 水平,减少AngⅡ的有害作用,从而增强心脏肾脏的保护作用,提示 ACEI和AT1R可成为COVID-19患者心肾损害的潜在治疗靶点。
现有技术中对拉萨大黄化学成分和药理活性进行了广泛的研究,发现拉萨大黄中主含有以曲札茋苷、甲基虎杖苷和虎杖苷等高含量二苯乙烯类化合物,特别是曲札茋苷具有潜在的医药用途。曲札茋苷化合名称为((E)-1-(3,5-二羟苯基)-2-(3-羟基-4-O-β-D-吡喃葡萄糖苯基)乙烯或3,5,3',4'-四羟基茋-3'-O-β-葡萄糖苷),其植物来源为拉萨大黄根茎,具体结构如下式I所示,白皮杉醇为曲札茋苷的苷元。研究表明,曲札茋苷具有抗炎、抗氧化、清除自由基等多种活性,中国专利(申请号:201010116358.2)《曲札茋苷在制备防治心脑缺血基本制剂中的应用及其制备方法》,中国专利(申请号: 201210198159.X)《一种茋类化合物在制备预防和治疗抑郁症药物中的用途》;中国专利(申请号:201210202957.5)《曲札茋苷在制备改善微循环障碍药物中的应用》公开了曲札茋苷在提取工艺、检测方法。现有技术表明,曲札茋苷具有防治缺血性心脑血管疾病、抗抑郁和改善微循环等多方面的活性,但未见在用于在抑制ACE活性的作用,亦未公开用于治疗和预防器官损伤等方面的作用,特别是新型冠状病毒肺炎引起的心肾等多器官损伤药物中的用途报道。
本发明通过实验发现拉萨大黄提取物,特别是曲札茋苷及其衍生物具有抑制ACE活性的作用,能够下调患者体内的ACE蛋白质水平来影响新冠病毒的易感性。曲札茋苷能够在病毒入侵后,迅速抓住 ACE2蛋白质,通过不断地分子刺激和相互作用,调节这种酶受体结合结构和序列,从而降低ACE活性,通过调节ACE/AngⅡ/AT1轴和 ACE2/Ang-(1-7)/MasR轴,抑制ACE,上调ACE2,以及靶向作用于ACE的药物有助于减缓COVID-19患者的心肾损伤。也就是说,曲札茋苷并不是能够抵抗病毒,而是主要通过调节ACE/AngⅡ/ AT1R轴和ACE2/Ang-(1-7)/MasR轴的平衡来发挥心脏和肾脏的保护作用,从而达到治疗的作用。ACE和ACE2是交互影响的,体内正向ACE-AngⅡ-AT1R轴和负向ACE2-Ang(1-7)-Mas轴之间相互制衡,保持血压及内环境的稳定。(Iwai M,Horiuchi M.Devil and angel in the renin-angiotensin sys-tem:ACE-angiotensinⅡ-AT1 receptor axis vs.ACE2-angiotensin-(1-7)-Mas receptor axis [J].Hypertens Res,2009,32(7):533-536.)
基于上述对SARS-CoV-2感染导致心肾损伤的机制分析可见,调节RAS系统的药物以及靶向抑制ACE的药物等可能有助于减缓 COVID-19患者的心肾损伤。但由于SARS-CoV-2与人类所见过的任何病原体都不一样,所以很难对它有一个清晰的认识,因此了解机体本身固有的病理生理机制,对于未来药物的研发同样重要,也是当前迫切需要解决的问题,对于临床治疗及药物研发具有潜在应用价值。
根据SARS-CoV-2造成心肾损伤的可能发病机制,曲札茋苷及其衍生物能针对以上发病机制进行干预,对SARS-CoV-2造成心肾损伤起到预防和治疗作用。
本发明的有效成分可以从植物中提取,产率高,可用于工业化生产,所用药物容易获得,市场前景好。
发明内容
本发明一方面,提供拉萨大黄提取物在制备用于预防和治疗器官损伤药物中的用途。
进一步的方案,所述的器官为心脏、肾脏,优选为肾脏。
进一步的方案,所述的用途是拉萨大黄提取物通过ACE活性抑制介导发挥的作用。
进一步的方案,所述的器官损伤是由病毒引起,更进一步所述的病毒为新冠肺炎病毒、SARS-CoV或MERS-CoV,优选为新冠肺炎病毒。
进一步的方案,所述的拉萨大黄提取物为曲札茋苷、白皮杉醇、白藜芦醇、虎杖苷、土大黄苷元-3’-O-β-D-葡萄糖苷、去氧土大黄苷,优选为曲札茋苷。
更进一步的方案,所述的药物还包括药学上可接受的辅料,更进一步可制备成常规的药物剂型,所述的剂型包括但是不具限于注射剂、片剂、胶囊剂、粉剂、丸剂或口服液。
本发明一方面,提供拉萨大黄提取物在制备通过抑制ACE活性介导疾病中的应用,其中所述的疾病不包括心脑血管疾病、神经系统疾病。
进一步的方案,所述的拉萨大黄提取物为曲札茋苷。
进一步的方案,所述的疾病为心脏、肾脏器官损伤。
具体地,曲札茋苷结构式如式I:
具体地,白皮杉醇结构式如式II:
具体地,白藜芦醇结构式如式III:
具体地,虎杖苷结构式如式IV:
具体地,土大黄苷元-3’-O-β-D-葡萄糖苷结构式如式V:
土大黄苷元-3’-O-β-D-葡萄糖苷
具体地,去氧土大黄苷结构式如式VI:
本发明发现曲札茋苷及其衍生物具有抑制ACE活性的作用,在介导抑制ACE活性的药物中具有潜在的作用;并在发挥通过调节 RAS系统的平衡以及靶向抑制ACE的药物有助于减缓COVID-19患者的心肾损伤,防治新冠患者后期引起的多器官衰竭的作用。该类成分可以从植物拉萨大黄中提取,产率高,可用于工业化生产,所用药物容易获得,市场前景好。但是本发明所述的拉萨大黄提取物成分并不局限于来源从植物拉萨大黄中的提取,任何通过化学合成手段获得的拉萨大黄提取物如曲札茋苷及其衍生物亦是本发明所要求的保护范畴。
附图说明
图1-1至1-10为本发明一种肾脏病理组织学检查图片(HE染色 20×)示意图,图中:
图1-1为正常对照组,基本正常,偶见肾皮质局部有少量瘀血;
图1-2为正常对照组,基本正常;
图1-3为模型组,肾小球囊腔缩窄,肾小球不易辨认,肾小管结构紊乱,间质里含有淋巴细胞群,皮质和髓质均可见多灶性蛋白管型,病变范围广,肾小管细胞扁平化,且多见细胞坏死;
图1-4为模型组,肾小球不易辨认,肾小管结构紊乱,间质里含有淋巴细胞群,皮质和髓质均可见多灶性蛋白管型,病变范围广,肾小管细胞扁平化,且多见细胞坏死;
图1-5为阳性对照组,肾小球病变明显改善,可见少数肾小球球囊粘连,系膜区轻度增生肾小管上皮细胞轻度局灶性增生,皮髓交界处偶见蛋白管型,肾小管细胞扁平化,未见细胞坏死;
图1-6为阳性对照组,皮髓皮质区弓形动脉周边有局灶性炎症,可见蛋白管型,肾小球病变明显改善;
图1-7为曲札茋苷低剂量组,肾小球病变明显改善,可见少数肾小球球囊粘连,系膜区轻度增生肾小管上皮细胞轻度局灶性增生,皮质可见蛋白管型,皮髓交界处弓形动脉周边局灶性炎症;
图1-8为曲札茋苷低剂量组,皮质可见蛋白管型;
图1-9曲札茋苷高剂量组,肾小球病变明显改善,可见少数肾小球球囊粘连,系膜区轻度增生肾小管上皮细胞轻度局灶性增生,肾小管间距增大,皮质、髓质、皮髓交界处均可见蛋白管型;
图1-10曲札茋苷高剂量组,少量蛋白管型。
具体实施方式
名词解释
如果没有其他特殊定义本文所述的ACE2一般是指受体血管紧张素转换酶(Angiotensin converting enzyme,ACE)2。
如果没有其他特殊定义本文所述的ACE一般是指血管紧张素转化酶,ACEI是指血管紧张素转化酶抑制剂。
下面结合说明书附图对本发明实施例作进一步详细描述。
实施例1:对ACE抑制作用研究
本实验采用蛋白序列为Abz-SDK(Dnp)P-OH的蛋白作ACE的底物,与ACE结合即释放荧光基团,通过酶标仪检测荧光生成的平均速度,即可反映ACE酶的活性。将受试物配成不同浓度,按表1加入反应体系,于波长460nm处测定平均反应速度,计算各受试物的抑制率。每浓度设置3孔重复,结果取均值。
(1)供试品
土大黄苷元-3`-O-β-D-glu:米白色粉末;
去氧土大黄苷:淡紫色结晶粉末;
白藜芦醇:白色细小结晶粉末;
虎杖苷:淡黄色粉末;
白皮杉醇:粉白色粉末;
曲札茋苷:白色粉末;
以上样品由昆药集团药物研究院提供。
(2)仪器及试剂:
A:仪器
HS-25型pH计,上海雷磁科学仪器厂;超声波清洗器,SK-8200H,上海科导超声仪器有限公司;SYNERGY HT酶标仪,BioTek;0.5~ 10单道微量移液器,Finnpipette;0.5~10ul单道微量电子移液器, 10~100ul单道微量移液器,10~300单道微量电子移液器,50~1000 单道微量电子移液器,Eppendorf。
B:试剂
成套pH校准缓冲剂,上海市爱建试剂厂出品,批号0920515; Tris-base,Purity≥99.9%,Amresco产品,Exp2012/12;浓HCl,云南汕滇药业有限公司,批号20090420;NaOH,云南汕滇药业有限公司,批号20091001;二甲基亚砜(DMSO):天津市科密欧化学试剂有限公司,批号:20110816;ACE(Angiotensin Converting Enzyme from rabbit lung):sigma,货号A6778 1UN;ACE荧光底物: Abz-SDK(Dnp)P-OH;卡托普利:sigma,lot#SLBF3962V;
纯化水,现取现用。
(3)试剂准备、标准和样品质量控制
A:试剂制备
1. 1M Tris-HCl(121.1)
取Tris-base 121.1g,加水800ml溶解,以浓HCl调节pH7.00,定容至1000ml,必要时121℃灭菌30min后使用,4℃保存。
2. 4*BUFFER
称取Tris-base 4.844g,加水80ml溶解,加入50mM的NaCl,以浓HCl调节pH8.2,定容至100ml。
3. 50mM ZnCl2溶液
称取67.5mg的ZnCl2,加入10ml去离子水,得到50mM的ZnCl2溶液。
4. 4mM ACE(SDK)底物储备液(731.72)
称取ACE底物2.0mg,加入683.3ul 1M Tris-HCl溶解,置4℃保存,临用前加Tris-HCl稀释至所需浓度,再加入50mM的ZnCl2溶液。
5.ACE原液
临用前取1un/ml的原液适量,以去离子水稀释至所需浓度并置于冰上待用。
6. 10mM卡托普利(217.29)
称取0.0017g卡托普利,加入782μl DMSO溶解,常温保存,临用前稀释至所需浓度。
B:对照品和样品的质量控制
ACE:贮存于-20摄氏度;
ACE(SDK)底物:贮存于-20℃,biomatik,lot#p131216- mj379828;
阴性对照:DMSO;
阳性对照:卡托普利;
样品:一般配置成10mM,检测低浓度活性时,在40min内完成检测。
(4)实验方法:
将不同浓度的曲扎茋苷系列样品配置成不同浓度,按下表中的反应体系加入,每个浓度置3孔重复,结果取其均数。加ACE开始反应后于酶标仪上测定,以平均反应速度判断ACE酶的活性。通过公式计算出检品对ACE抑制率。
表1 ACE抑制剂测定体系
ACE抑制率计算:
抑制率(%)=((△A(标准空)-△A(样品空))/△A(标准空)))x100%
△A:扣除本底后的平均反应速度均值
五、结果和结论:
根据实验数据作图计算出IC50,实验结果见表2。
表2系列样品对ACE的抑制作用
由表2可见,阳性对照卡托普利抑制ACE的IC50为1.6nM,与文献报道相近(1.4nM),表明该体系稳定;6个受试物体外对ACE均有不同程度的抑制作用,其IC50分别为0.27、23.38、6.23、2.85、1.08、 15.75μM,抑制强度顺序为:曲札茋苷>白皮杉醇>白藜芦醇>虎杖苷> 土大黄苷元-3’-O-β-D-葡萄糖苷>去氧土大黄苷,表明曲札茋苷对ACE的抑制活性最强。
实施例2:对肾炎大鼠的影响
(1)实验材料
1.1样品
1.1.1受试物
注射用曲札茋苷:20mg/支,昆药集团药物研究院提供。
1.1.2阳性对照
盐酸贝那普利片(洛汀新),国药准字H20030514,10mg/片,北京诺华制药有限公司生产,批号X1774;实验时,用1%CMC-Na配制成0.6mg/mL浓度的混悬液。
1.1.3溶媒对照
羧甲基纤维素钠(CMC-Na),化学纯,500g/瓶,国药集团化学试剂有限公司,批号F20100909,用纯水加热、煮沸、冷却后配成1%浓度,经高压灭菌后供受试样品配制及溶媒对照用,一次配制,使用 5~7天。
1.2动物
SPF级雄性SD大鼠,6~8周龄,体重200~250g;SPF级昆明种小鼠,体重18~20g,雌雄各半;均由昆明医学院实验动物中心提供,生产许可证:SCXK(滇)2011-0004,发证机关:昆明市科学技术局;饲养于IVC动物实验室,温度20~25℃(日温差≤3℃),湿度40%~70%,照明12h:12h明暗交替,照度150~300lx,噪音≤60dB,实验动物使用许可证:SYXK(滇)2014-0001,发证单位:昆明市科学技术局;PVC透明塑料盒群养,每箱≤6只,每日喂饲鼠用配合饲料,自由饮水,视情况更换笼具和垫料,饲料来源于昆明医学院实验动物中心,生产许可证:SCXK(滇)2011-0004,发证单位:昆明市科学技术局。
1.3主要仪器
AL104型电子分析天平,梅特勒-托利多仪器(上海)有限公司;JJ2000型电子天平,常熟双杰测试仪器厂;F5810-R型台式高速冷冻离心机,德国Eppendorf公司;Thermo1500型酶标仪,芬兰雷勃公司;Millipore超纯水处理系统,美国Millipore公司;MVIS-2015 型全自动血液流变仪,重庆天海;XW-80A型漩涡混合器,上海精科实业有限公司;BCD-193冰箱,合肥荣事达电冰箱有限公司;725型超低温冰箱,美国Thermo公司;HHS-1型恒温水浴箱,金坛市大地自动化仪器厂。
1.4主要试剂
1.4.1造模试剂
注射用盐酸多柔比星(盐酸阿霉素),国药准字H44024359,10mg/ 瓶,深圳万乐药业有限公司,批号0806E1,2~8℃保存,临用时用氯化钠注射液配成1mg/mL。
1.4.2检测试剂
尿蛋白定量试剂盒(CBB法),96T/份,南京建成生物工程研究所,批号20140621;大鼠肿瘤坏死因子(TNF-a)、前列腺素E2(PGE2) 酶联免疫检测试剂盒,批号均为201407,购自R&D公司;谷胱甘肽 (GSH),南京建成生物工程研究所,批号20141206;SOD试剂盒(黄嘌呤氧化法),96T/份,南京建成生物工程研究所,批号20141205; MDA试剂盒(硫代巴比妥酸法),96T/份,南京建成生物工程研究所,批号20141208;NO试剂盒(一步法),96T/份,南京建成生物工程研究所,批号20140827。
1.4.3其他试剂
氯化钠注射液,500ml/瓶,贵州天地药业有限责任公司,批号 A11102207;肝素钠,规格500mg/支,批号85810250,北京鼎国生物技术有限责任公司。
(2)实验方法
2.1模型制备与分组给药
将250~300g雄性SD大鼠60只在IVC动物房适应性喂养10天,除留10只为正常对照外,其余50只用于造模,分别于1天、7天、14天按 3、2、1mg/kg的剂量给每只大鼠尾静脉注射0.1%盐酸阿霉素溶液,正常对照组注射等容量的生理盐水。造模21天收集尿液,测定24小时尿蛋白,结果显示21天尿蛋白普遍升高,表明模型成功。将造模成功的 43只大鼠,按24h尿蛋白总量和体重均匀分为4组:模型对照组13只、洛丁新组10只、注射用曲札茋苷高、低剂量组各10只。各组动物每日按剂量腹腔给药一次,连续56d,容量均为10mL/kg.bw,正常和模型对照组给予等容量1%CMC-Na。每周称重1次,根据体重调整给药量。
2.2观察指标及测定
2.2.1一般情况观察
造模后每天观察大鼠进食和活动状态,每天称取体重一次。
2.2.2尿蛋白定量
分别收集每只动物造模21天及给药后7、14、21、28……56天的 24h尿液,量取体积,用CBB法测定尿液中蛋白含量,计算尿蛋白总量,以mg/24h表示。
2.2.3血液生化及抗氧化指标检测
末次给药后禁食不禁水12小时,麻醉动物后,股动脉取血4ml于肝素抗凝管中,4℃、4000rpm离心10min,分离血清,用全自动生化分析仪测定血清TP、ALB、GLO、Cre、BUN、TC、TG、AST、ALT 水平,并按试剂盒方法测定NO、SOD、MDA含量。
2.2.4脏器系数测定
取血后脱颈椎处死大鼠,剖取胸腺、脾脏、肾、肾上腺称重,计算各脏器系数。脏器系数=脏器质量(mg或g)/动物体重(g)×100
2.2.5肾组织匀浆相关指标检测
将右肾置于-70℃冰箱保存。测定前,取同一部位右肾组织200mg 于玻璃匀浆器中,按1∶9加入冰生理盐水,冰浴匀浆,4000r/min、4℃离心5min。取上清液,用ELISA方法测定TNF-α、PGE2含量,并按试剂盒方法测定SOD、MDA水平。
2.2.6组织病理学观察
将左肾放入10%的甲醛溶液固定,经梯度乙醇脱水,二甲苯透明,浸蜡包埋,常规切片,HE染色,中性树胶封片,光学显微镜下观察,盲法阅片。肾脏组织切片评分标准见表3。
表3肾脏组织切片(HE染色)评分标准
2.3统计学处理
资料采用Ridit检验;P<0.05有统计学意义,P<0.01有显著统计学意义。
(3)实验结果
3.1对体重的影响
与对照组相比:▲/▲▲P<0.05/0.01
由表4可见,给药前各模型组动物基础体重与正常对照组相比无明显差异;至给药结束,除正常对照组动物体重增长明显外,其余各组体重增长缓慢,但与模型对照组相比无统计学差异。
3.2对模型大鼠尿蛋白的影响
与对照组相比:▲/▲▲P<0.05/0.01;与模型组相比:*/**P<0.05/0.01
3.3对血液相关指标的影响
3.3.1对血液生化的影响
由表6可见,与正常对照组相比,模型组动物血清TP、ALB显著下降,BUN、Cre、TG、TC显著升高,ALT、AST有降低趋势,GLO 无明显变化,表明阿霉素肾炎大鼠继发高脂血症和低蛋白血症;各组连续给药8周,均能明显升高血清ALB含量及A/G,明显降低BUN、 Cre、TC,对TP均有回升趋势;除曲札茋苷高剂量组对TG仅有降低趋势外,所有给药组均能明显降低血清TG水平;除阳性对照组对 AST、ALT有明显升高作用外,曲札茋苷两剂量组对AST和ALT无明显影响。提示阳性对照及曲札茋苷能明显对抗阿霉素肾炎模型大鼠继发的高脂血症和低蛋白血症。
与对照组相比:▲/▲▲P<0.05/0.01;与模型组相比:*/**P<0.05/0.01
3.3.2对NO、MDA和SOD表达的影响
由表7可见,模型对照组血清NO水平和MDA含量明显升高, SOD明显下降,与对照相比有统计学意义(p<0.05/0.01);所有给药组均明显降低模型动物血清NO、MDA水平;不同程度升高SOD活力,其中阳性对照、曲札茋苷高剂量组的作用接近统计学差异 (p=0.06)。
与对照组相比:▲/▲▲P<0.05/0.01;与模型组相比:*/**P<0.05/0.01
3.4对脏器指数的影响
由表8可见,与正常对照组相比,模型组肾上腺、肾脏和脾脏指数明显增加(p<0.01),胸腺指数呈降低趋势;与模型对照组相比,各给药组能明显降低肾上腺及肾脏指数(P<0.05),不同程度降低脾脏指数,对胸腺指数有升高趋势。
与对照组相比:▲/▲▲P<0.05/0.01;与模型组相比:*P<0.05
3.5对肾组织匀浆相关指标的影响
由表9可见,与正常对照组相比,模型组肾组织匀浆中TNF-α和MAD含量明显增加(p<0.01/0.05),SOD及PGE-2则无明显变化;各给药组均能明显降低模型动物肾组织匀浆中TNF-α含量(p<0.01),但对MDA无明显影响。
与对照组相比:▲/▲▲P<0.05/0.01;与模型组相比:**P<0.01
3.6组织病理学观察
3.6.1普通光镜检查
图1为肾脏病理组织学检查图片(HE染色20×),曲札茋苷两剂量组病变程度介于模型组和阳性药组之间,病变程度有所减轻。
3.6.2半定量评分
由表10可见,与正常对照组相比,模型组动物肾脏组织病理变化有显著差异(P<0.01);与模型组相比,各给药组均能明显减轻模型大鼠肾脏组织的病理学改变(P<0.01/0.05)。
表10对模型大鼠肾脏组织病理状况的影响
曲札茋苷以5、10mg/kg剂量连续腹腔给药8周,能明显降低阿霉素肾炎模型大鼠的蛋白尿、提高血清ALB和A/G、降低BUN和Cre,对TC、TG有明显降低作用。提示其能纠正肾炎大鼠的低蛋白血症、高脂血症状态,明显改善肾功能,并能明显减轻模型大鼠肾脏组织的病理学改变;曲札茋苷连续给药8周,能明显降低模型大鼠血清NO、 MDA含量,不同程度增加SOD的活力,明显降低肾脏组织中TNF-α的表达,提示可减轻肾脏组织的损伤。
显然,本领域的技术人员可以对本发明进行各种改动和变型而不脱离本发明的精神和范围。这样,倘若本发明的这些修改和变型属于本发明权利要求及其等同技术的范围之内,则本发明也意图包含这些改动和变型在内。
Claims (10)
1.一种拉萨大黄提取物在制备用于预防和治疗器官损伤药物中的用途。
2.根据权利要求1所述的用途,其特征在于所述的器官为心脏、肾脏,优选为肾脏。
3.根据权利要求1所述的用途,其特征在于拉萨大黄提取物通过ACE活性抑制介导发挥的作用。
4.根据权利要求1所述的用途,其特征在于所述的器官损伤是由病毒引起,所述的病毒为新冠肺炎病毒、SARS-CoV或MERS-CoV,优选为新冠肺炎病毒。
5.根据权利要求1所述的用途,其特征在于所述的拉萨大黄提取物为曲札茋苷、白皮杉醇、白藜芦醇、虎杖苷、土大黄苷元-3’-O-β-D-葡萄糖苷、去氧土大黄苷。
6.根据权利要求5所述的用途,其特征在于拉萨大黄提取物为曲札茋苷。
7.根据权利要求1-6所述的用途,其特征在于所述的药物还包括药学上可接受的辅料,制备成常规的药物剂型,所述的剂型包括注射剂、片剂、胶囊剂、粉剂、丸剂或口服液。
8.一种拉萨大黄提取物在制备通过抑制ACE活性介导疾病中的应用,其中所述的疾病不包括心脑血管疾病、神经系统疾病。
9.根据权利要求8所述的用途,所述的拉萨大黄提取物为曲札茋苷。
10.根据权利要求8所述的用途,所述的疾病为心脏、肾脏器官损伤。
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