【作者】 周瑞芳；

【导师】 刘鹏熙；

【作者基本信息】 广州中医药大学 ， 中医外科学， 2008， 博士

【摘要】 研究背景和目的乳腺癌是严重危害妇女健康的恶性肿瘤,在女性恶性肿瘤中其发病率和死亡率均居首位。尽管我国为乳腺癌低发国家,但在我国一些大城市乳腺癌发病率增长很快,已跃居女性恶性肿瘤的首位,成为妇女健康的重大威胁。在国内,中药普遍用于乳腺癌的辅助治疗,随着替代医学的兴起,欧美和其他西方国家应用中药辅助治疗肿瘤的兴趣逐渐增大。同时激素替代疗法（HRT）可增加绝经后妇女乳腺癌危险性,因此,天然药物的应用越来越受到重视。传统中药中,不少方、药存在植物雌激素（phytoestrogen,PE）效应,在含有PE成分的中药中,活血化瘀类和补益类中药占的比例居各类中药的前二位。黄芪为豆科多年生草本植物蒙古黄芪或膜荚黄芪的根,具有补气升阳,益卫固表,利水消肿,托疮生肌的功效,富含黄酮和异黄酮,它是具有雌激素样活性的有效成分。黄芪被广泛应用于肺癌、胃癌、乳腺癌的辅助治疗,以减轻化疗药物的毒副反应,协同化疗减毒增效,治疗放疗、化疗引起的骨髓抑制,、提高机体的免疫功能和改善生活质量。相关实验研究大都提示黄芪或其提取物有一定抗肿瘤作用,可抑制各种肿瘤细胞株的生长,诱导细胞凋亡。但也有实验发现黄芪或其提取物促进肿瘤细胞增殖。丹参为活血类代表药,具有活血化瘀调经,凉血消痈,安神的功效。脂溶性成分丹参酮、丹参醌药理作用与植物性雌激素相似。多年来其各种制剂被广泛应用于肿瘤尤其是乳腺癌的辅助治疗。相关实验研究以及临床应用表明,单味药丹参、或丹参的提取物、或丹参与其他药物组成的复方制剂均具有一定的抗肿瘤作用。但有实验表明丹参可促进实验动物的肿瘤转移。植物雌激素是非甾类、植物来源的具有雌激素活性的天然化合物,其结构与雌激素非常近似。目前已知的PE主要包括以下3类:异黄酮类、木脂素类和香豆雌酚。PE与ER结合后的雌激素效应较雌二醇降低10 000—14 000倍,又竞争占据ER,低剂量的植物雌激素表现为雌激素拮抗效应,中剂量时可产生一定的雌激素活性,高剂量时补充的异黄酮可以活化因雌激素水平限制未能活化的ER,产生雌激素增强效应。全球乳腺癌发病的地域分布状况显示,亚洲的发病率最低。作为亚洲人民经常食用的黄豆及其制品受到关注,黄豆的防癌作用主要和异黄酮有关。高血浆水平的genistein与乳腺癌发病的低风险相关。植物雌激素与雌激素受体结合后,促进性激素结合球蛋白产生,抑制酪氨酸激酶和雌激素合成酶的活性,降低靶器官中性激素的生物活性,从而减少癌症的发生。但国外也有学者认为PE对激素依赖性肿瘤乳腺癌的有效性和安全性并没有循证医学的证据,在作为化学预防剂的同时又能促进激素受体阳性肿瘤细胞生长,甚至拮抗TAM的抗肿瘤增殖作用,因此PE摄入对激素依赖性肿瘤可能产生不良作用,对肿瘤患者和肿瘤生存者是高风险的。一些植物雌激素样中药,它们或与他莫昔芬,或与芳香化酶抑制剂相互作用产生不利影响,潜在地增加乳腺癌的危险性。植物雌激素对激素依赖性乳腺癌的确切效应尚存在争议,尚缺少有关PE在乳腺癌患者及同时服用三苯氧胺者中使用合理性的确凿依据。因此,本实验选用临床应用较多,研究较多,争议较大,植物雌激素效应确切的代表性药物黄芪、丹参;选用激素依赖性肿瘤乳腺癌两种激素受体不同的细胞株MCF-7（ER+）、MDA-MB-231（ER-）进行研究。通过研究丹参、黄芪在生理剂量E2环境及药理剂量他莫昔芬（Tamoxifen TAM）环境下对MCF-7、MDA-MB-231细胞增殖及凋亡的影响,希望能够为植物雌激素类中药在乳腺癌的辅助治疗中的安全性和有效性寻找更丰富的实验室依据及理论基础。第一部分预实验:黄芪、丹参注射液对人乳腺癌MCF-7细胞增殖的影响目的用MTT法筛选黄芪、丹参注射液作用于人乳腺癌MCF-7细胞的最佳实验浓度材料和方法ER（+）人乳腺癌MCF-7细胞以RPMI1640（10%小牛血清,青霉素100U/mL,链霉素100ug/mL）培养,选用对数生长期MCF-7细胞,以1×10e4/ml细胞浓度接种于96孔板,设置细胞对照组、各药物浓度组。MTT法检测各浓度的黄芪、丹参对MCF-7细胞增殖的影响。OD值以均数±标准差表示,组间比较用方差分析。以P＜0.05表示差异有统计学意义。结果不同浓度的黄芪注射液作用于人乳腺癌MCF-7细胞48h后,与细胞对照相比,对MCF-7细胞的增殖均有抑制作用（P＜0.05）。药物浓度大于2×10^-5g/ml时,剂量依赖性明显;药物浓度小于2×10^-5g/ml时,其效应曲线呈不规则形。不同浓度的丹参作用于MCF-7细胞后,与细胞对照相比,1×10^-2g/ml、1×10^-6g/ml的浓度对MCF—7细胞有抑制作用（P＜0.05）;其他各浓度对细胞增殖无明显影响。1×10^-5g/ml—1×10^-2g/ml的剂量其对MCF-7细胞生长的影响有线性关系;小于1×10^-5g/ml时,其效应曲线呈不规则形。结论本实验黄芪注射液选用2×10^-1g/ml—2×10^-5g/ml五个浓度,丹参注射液选用1×10^-1g/ml—1×10^-5g/ml五个浓度进行进一步的研究。第二部分黄芪注射液对人乳腺癌细胞株MCF-7、MDA-MB-231增殖及凋亡的影响研究目的探讨具有植物雌激素效应的黄芪注射液在生理剂量E2环境下及药理剂量他莫昔芬环境下对雌激素受体阳性（ER+）人乳癌细胞株MCF-7、雌激素受体阴性（ER-）人乳腺癌细胞株MDA-MB-231增殖及凋亡的影响。材料和方法ER（+）人乳腺癌MCF-7细胞、ER（-）人乳腺癌MDA-MB-231细胞以RPMI1640（10%小牛血清,青霉素100U/mL,链霉素100ug/mL）培养。设置细胞对照组、TAM对照组。观察各浓度黄芪注射液在生理剂量E2环境及药理剂量TAM环境下对MCF-7、MDA-MB-231细胞生长的影响。MTT法检测细胞增殖,流式细胞仪分析细胞周期,流式细胞仪检测凋亡率、DAN ladder观察凋亡。所有资料以均数±标准差表示,计量资料用方差分析,计数资料用卡方检验。以P＜0.05表示差异有统计学意义。结果生理剂量E2环境下,各种剂量的黄芪注射液在各个作用时间点均无促进MCF-7细胞增殖的效应,随着作用时间的延长,48h、72h各剂量均呈现抑制MCF-7细胞增殖的效应（P＜0.05）,72h其作用均较单用TAM强（P＜0.05）;高剂量(2×10^-1g/ml)作用72h促进MCF-7细胞凋亡,凋亡率为16.7%。与TAM合用,各剂量的黄芪注射液在24h、48h、72h均呈现抑制MCF-7细胞增殖的效应（P＜0.05）,呈剂量依赖性,无时间依赖性;高剂量(2×10^-1g/ml)作用72h促进MCF-7细胞凋亡,凋亡率为22.1%。两种环境下,黄芪注射液作用24h后,高剂量(2×10^-1g/ml)增加MCF-7细胞S期细胞比例,降低G0/G1、G2/M期细胞比例（P＜0.05）;中高剂量(2×10^-2g/ml)增加MCF-7细胞G0/G1期细胞比例,降低S、G2/M期细胞比例（P＜0.05）。中、中低剂量(2×10^-3g/ml、2×10^-4g/ml)在生理剂量E2环境下增加MCF-7细胞G0/G1、S期细胞比例,降低G2/M期细胞比例（P＜0.05）。对于ER（-）MDA-MB-231细胞,黄芪注射液高剂量(2×10^-1g/ml)抑制其增殖（P＜0.05）,促进其凋亡,而对其细胞周期无明显影响;中高、中剂量对其增殖影响与时间点相关,对其凋亡无影响,使细胞进入S期,增加了其DNA含量（P＜0.05）;中低、低剂量组促进人乳腺癌MDA-MB-231细胞的增殖（P＜0.05）,对其细胞周期和凋亡率均无明显影响。结论临床上黄芪在雌激素受体阳性的乳腺癌及应用TAM患者中的应用是安全的,且一定剂量范围内可抑制雌激素受体阳性的乳腺癌细胞增殖,诱导其凋亡,阻滞细胞的有丝分裂于S或G2/M期,因此对这部分患者可能是有益的。高剂量黄芪抑制人乳腺癌MDA-MB-231细胞增殖,诱导其凋亡,提示临床上高剂量的黄芪在ER（-）乳腺癌患者中的使用是安全和有益的;低、中低剂量黄芪促进MDA-MB-231细胞增殖;中高剂量使MDA-MB-231细胞G0/G1期细胞比例下降,S期细胞比例上升,可能促进了其有丝分裂。一定剂量范围内黄芪可能促进雌激素受体阴性的乳腺癌细胞的生长,其在临床乳腺癌患者中使用的剂量范围及安全性尚待进一步体内实验研究。第三部分丹参注射液对人乳腺癌细胞株MCF-7、MDA-MB-231增殖及凋亡的影响研究目的探讨具有植物雌激素效应的丹参注射液在生理剂量E2环境下及药理剂量他莫昔芬环境下对雌激素受体阳性（ER+）人乳癌细胞株MCF-7、雌激素受体阴性（ER-）人乳腺癌细胞株MDA-MB-231增殖及凋亡的影响。材料和方法ER（+）人乳腺癌MCF-7细胞、ER（-）人乳腺癌MDA-MB-231细胞以RPMI1640（10%小牛血清,青霉素100U/mL,链霉素100ug/mL）培养。设置空白对照组、TAM对照组。观察各浓度丹参注射液在生理剂量E2环境及药理剂量TAM环境下对MCF-7、MDA-MB-231细胞生长的影响。MTT法检测细胞的增殖,流式细胞仪分析细胞周期,流式细胞仪检测凋亡率、DAN ladder观察凋亡。所有资料以均数±标准差表示,计量资料用方差分析,计数资料用卡方检验。以P＜0.05表示差异有统计学意义。结果生理剂量E2环境下,高剂量(1×10^-1g/ml)丹参注射液作用24h、72h促进MCF-7细胞增殖（P＜0.05）,作用48h对MCF-7细胞的增殖无影响。低于1×10^-1g/ml的剂量,抑制MCF-7细胞增殖（P＜0.05）,其效应与剂量反相关,无时间依赖性。与TAM合用,高剂量丹参注射液(1×10^-1g/ml)促进MCF-7细胞增殖的效应得到不同程度抑制,24h促增殖率下降,72h对增殖无影响（P＞0.05）。其他各剂量抑制MCF-7细胞增殖（P＜0.05）。两种环境下,丹参注射液低剂量(1×10^-5g/ml)抑制MCF-7细胞增殖的效应均较TAM强（P＜0.05）;高剂量(1×10^-1g/ml)诱导MCF-7细胞凋亡,各剂量对细胞周期均无明显影响。对于ER（-）人乳腺癌MDA-MB-231细胞,丹参注射液作用48h抑制MDA-MB-231细胞增殖（P＜0.05）,作用72h促进细胞增殖（P＜0.05）,其效应均有剂量依赖性。作用24h其效应和浓度相关,高剂量(1×10^-1g/ml)促进增殖（P＜0.05）,低剂量(1×10^-5g/ml)抑制增殖（P＜0.05）,其他各剂量对细胞生长无影响。1×10^-1g/ml、1×10^-2g/ml丹参注射液诱导MDA-MB-231细胞凋亡;中低剂量(1×10^-4g/ml)降低MDA-MB-231细胞G0/G1期DNA含量,增加S、G2-M期细胞DNA含量。结论丹参对于雌激素受体阳性的人乳腺癌MCF-7细胞的作用方式类似植物雌激素,高剂量促进MCF-7细胞增殖,低剂量抑制其增殖,其双向调节作用与剂量相关,在临床使用的安全性、有效性及剂量范围,需要进一步实验研究。丹参注射液对ER（-）人乳腺癌MDA-MB-231细胞生长的影响与时间点、细胞系、药物浓度均相关。中低剂量可能通过部分活化雌激素受体β（ERβ）增加S、G2-M期细胞比例而促进有丝分裂。提示临床上丹参在雌激素受体阴性患者中的使用需谨慎,其安全性尚需进一步在体实验研究。更多还原

【Abstract】 Background and objectiveBreast cancer is one of the malignant tumour which hazards women’s health seriously.Breast cancer incidence and mortality vary considerably by world regions.In general,the incidence is high（greater than 80 per 100,000）in developed regions of the world and low（less than 30 per 100,000）,though increasing,in developing regions;it increasing sharply in some big cities of China.Chinese herb was managed as one kind of adjunctive therapy of breast cancer.As rising of replacement medicine and hormone replacement therapy（HRT）increasing risk of breast cancer in post-menopause women,chinses herb was managed to treat tumor and climacteric syndrome frequently in other countries.The effects of some decoction and herb were similar to phytoestrogens,herb to active blood and tonify are the most common.Astragalus mongholicus is rich in flavone and isoflavone,which are the effective component of estrogenic effect.Astragalus mongholicus was generally applied to lung cancer、gastric carcinoma、breast cancer to relieve adverse reaction and in coordination with chemotherapy,treat myelosuppression,boost immunological function and improve quality of life.Related studies suggested that Astragalus mongholicus or its extracts could inhibit proliferation and induce apoptosis of some tumor cell lines.But others discovered Astragalus mongholicus may promote tumor cell growth.Salvia miltiorrhiza is the representative medicine to active blood, the pharmacologic action of tanshinone,the liposoluble constituent of salvia miltiorrhiza,is resemble to phytoestrogen.Various kinds praeparatum were managed as adjunctive therapy of mammary adenocarcinoma widespread.Some study and clinical application indicated that Salvia miltiorrhiza,its extract,its compound preparation all had effect to inhibit tumor,simultaneously some found Salvia miltiorrhiza could promote tumor metastasis of experimental animals.Phytoestrogens（PE）are a group of plant-derived substances that are structurally or functionally similar to estradiol,concluding isoflavones、Lignans and coumestans.The estrogenic effects of phytoestrogen is 10 000 to 14 000 fold less potent than 17beta-estradiol （E2）after binding with estrogen receptor（ER）.As occupying ER competive, low concentration PE antagonize estrogen,middle dose have some estrogenic activity,high dose display phytoestrogenic activity.Interest in phytoestrogens has been fueled by epidemiologic data that suggest a decreased risk of breast cancer in women from countries with high phytoestrogen consumption.High genistein circulation levels are associated with reduced breast cancer risk.However,some foreigen scholar suggested PE could potentially increase the risk of breast cancer or interact with tamoxifen or aromatase inhibitors.Phytoestrogens may serve as chemopreventive agents while at the same time being capable of promoting growth in estrogen receptor positive cancer cell lines. Phytoestrogen intake might be ill advised for patients at an increased risk for hormone-dependent cancers,cancer patients,or cancer survivors. There is a paucity of clinical trial data of Some Chinese herb similar to PE demonstrating either safety or efficacy in patients with breast cancer or those with tamoxifen coadministration.The purpose of this study was to investigate the efficacy and safety of Astragalus mongholicus and Salvia miltiorrhiza in hormone sensitive （MCF-7）and insensitive（MDA-MB-231）breast cancer cell lines with physiological dose E2 or pharmacologic dose TAM. PartⅠPreliminary experiment:Effect of Astragalus mongholicus and Salvia miltiorrhiza on proliferation of human breast cancer cells MCF-7 in vitroObjectiveThe aim of the study was to find the optimal experimental concentration of Astragalus mongholicus and Salvia miltiorrhiza on human breast cancer cells MCF-7 with MTT assay.Materials and methodsHormone sensitive breast cancer MCF-7 cells were cultured in RPMI1640（10%calf serum,penicillin 100U/mL,phytomycin 100ug/mL）,MCF-7 cells in logarithmic growth phase inoculated in 96 well mask at 1×10e4/ml concentration,MTT assay was used to evaluate the proliferation of MCF-7 cells of Astragalus mongholicus or Salvia miltiorrhiza at kinds of concentrations.Optical density data were expressed as mean±standard deviation（SD）,and a one-way ANOVE was performed for group comparison.All statistical tests were two-sided and P＜0.05 was considered to indicate statistical significance.ResultsCompared with blank group,Astragalus mongholicus could inhibit the proliferation of MCF-7 cells at any concentration（P＜0.05）.Dose exceeded 2×10^-5g/ml,the effect was dose dependent;dose smaller than 2×10^-5g/ml, the effective curve was irregular.Salvia miltiorrhiza at concentration of 1×10^-2g/ml and 1×10^-6g/ml could inhibit the proliferation of MCF-7 cells（P＜0.05）,Others dose was uneffective.There is linear relationship when dose from 1×10^-5g/ml to 1×10^-2g/ml.ConclusionIn this study Astragalus mongholicus was selected at concentration from 2×10^-1g/ml to 2×10^-5g/ml;Salvia miltiorrbiza was selected at concentration from 1×10^-1g/ml to 1×10^-5g/ml.PartⅡEffect of Astragalus mongholicus on proliferation and apoptosis in hormone sensitive（MCF-7）and insensitive（MDA-MB-231）breast cancer cell linesObjective The aim of the study was to investigate the effect of Astragalus mongholicus on proliferation and apoptosis in hormone sensitive（MCF-7） and insensitive（MDA-MB-231）breast cancer cell lines with physiological dose E2 or pharmacologic dose TAM.Materials and methodsHormone sensitive（MCF-7）and insensitive（MDA-MB-231）breast cancer cells were cultured respectively in RPMI1640（10%calf serum,penicillin 100U/mL,phytomycin 100ug/mL）,the effects of different dose of Astragalus mongholicus on proliferation and apoptosis was observed in hormone sensitive（MCF-7）and insensitive（MDA-MB-231）breast cancer cell lines with physiological dose E2 or pharmacologic dose TAM,the proliferation of MCF-7 and MDA-MB-231 cells were evaluated with MTT assay,cell apoptotic rate were measured with flow cytometry and DNA ladder,cell cycle were observed by flow cytometry.Data were expressed as mean±standard deviation（SD）,and analysis of variance was performed for group comparison of measurement data,chi square test was used for enumeration data.All statistical tests were two-sided and P＜0.05 was considered to indicate statistical significance.ResultsWith physiological dose E2,Astragalus mongholicus did not promot MCF-7 cells proliferation at any concentration.As time lasting, Astragalus mongholicus showed better inhibitory effect than TAM（P＜0.05）.At 2×10^-1g/ml concentration Astragalus mongholicus increased the rate of apoptosis to 16.7%.Combined with TAM,Astragalus mongholicus showed dose dependent inhibitory effect on MCF-7 cells（P＜0.05）.The rate of apoptosis is 22.1%at 2×10^-1g/ml concentration.After cocultured 24 hours,Astragalus mongholicus significantly increased the proliferative phase（percent S-phase）and decreased G0/G1 andG2/M phase（P＜0.05）at high concentration(2×10^-1g/ml)with physiological dose E2 or combined with TAM,at 2×10^-2g/ml concentration,it accumulating cells in G0/G1 and G2/M phase,decreasing the proliferative phase（percent S-phase）（P＜0.05）.The influence was different in insensitive（MDA-MB-231）breast cancer cell lines at distinctive concentration.Astragalus mongholicus inhibited cell proliferation and increasing the rate of apoptosis at high concentration(2×10^-1g/ml);inhibited cell proliferation and increased the proliferative phase（percent S-phase）at 2×10^-2g/ml、2×10^-3g/ml concentration;but promoted MDA-MB-231 cell proliferation at 2×10^-4g/ml、2×10^-5g/ml concentration.ConclusionThe findings suggested it was safety for patients with breast cancer or administered TAM to use Astragalus mongholicus clinically.Astragalus mongholicus inhibit MCF-7 cell proliferation in coordination with TAM in the initial stage,it may be beneficial for patients with estrogen receptor positive by inhibiting MCF-7 cell proliferation and inducing apoptosis at some dose limit.Astragalus mongholicus is safe and potent to hormone insensitive breast cancer patients at high concentration by increasing the rate of apoptosis and inhibiting MDA-MB-231 cell growth.However,Astragalus mongholicus promoted MDA-MB-231 cell growth at low and less middle concentration,had adverse effect on the cell cycle by significantly decreased the resting phase G0/G1 phase)and increased the proliferative phase（percent S-phase）at less high concentration.Additional research on safety and quantitation of Astragalus mongholicus in serum,their interactions with plasma and cellular proteins,and their uptake in target tissues is necessary.PartⅢEffect of Salvia miltiorrhiza on proliferation and apoptosis in hormone sensitive（MCF-7）and insensitive（MDA-MB-231）breast cancer cell linesObjectiveThe aim of the study was to investigate the effect of Salvia miltiorrhiza on proliferation and apoptosis in hormone sensitive（MCF-7） and insensitive（MDA-MB-231）breast cancer cell lines with physiological dose E2 or pharmacologic dose TAM.Materials and methodsHormone sensitive（MCF-7）and insensitive（MDA-MB-231）breast cancer cells were cultured respectively in RPMI1640（10%calf serum,penicillin 100U/mL,phytomycin 100ug/mL）,the effects of different dose of Salvia miltiorrhiza on proliferation and apoptosis was observed in hormone sensitive（MCF-7）and insensitive（MDA-MB-231）breast cancer cell lines with physiological dose E2 or pharmacologic dose TAM,the proliferation of MCF-7 and MDA-MB-231 cells were evaluated with MTT assay,cell apoptotic rate were measured with flow cytometry and DNA ladder,cell cycle were observed by flow cytometry.Data were expressed as mean±standard deviation（SD）,and analysis of variance was performed for group comparison of measurement data,chi square test was used for enumeration data.All statistical tests were two-sided and P＜0.05 was considered to indicate statistical significance.ResultsWith physiological dose E2,Salvia miltiorrhiza at high concentration(1×10^-1g/ml)promoted MCF-7 cells proliferation at 24 and 72h（P＜0.05）,had no influence at 48h.Salvia miltiorrhiza had inverse association on dose when inhibiting MCF-7 cell growth at other less concentration（P＜0.05）.Combined with TAM,the promoting effectof high dose Salvia miltiorrhiza was negated at different degree,the promoting growth rate decreased at 24h,there was no promoting effect at 72h.Salvia miltiorrhiza inhibited MCF-7 cells proliferation at other lower dose （P＜0.05）.The inhibitory effect was stronger than TAM at low concentration(1×10^-5g/ml)combined with TAM or not（P＜0.05）.Salvia miltiorrhiza had no influence on MCF-7 cell cycle and induced cell apoptosis at high concentration(1×10^-1g/ml).The effect of Salvia miltiorrhiza inhibited MDA-MB-231 cell proliferation at 48h and promoted MDA-MB-231 cell proliferation at 72h was dose dependent.At 24h it inhibited growth at low dose(1×10^-5g/ml)but promoted proliferation at high dose(1×10^-1g/ml)（P＜0.05）.Astragalus mongholicus increased MDA-MB-231 apoptosis rate at 1×10^-1g/ml、1×10^-2g/ml,but increased the proliferative phase（percent S and G2/M phase）and decreased the resting phase（G0/G1 phase）at 1×10^-4g/ml（P＜0.05）ConclusionThe data suggested effects of Salvia miltiorrhiza on MCF-7 cells was similar to phytoestrogen,it promoting hormone sensitive（MCF-7）breast cancer cell growth at high concentration and inhibiting cell proliferation at lower concentration.Salvia miltiorrhiza is a class of potent phytoestrogen.The safety and quantitation of Salvia miltiorrhiza used in clinic need further research.Salvia miltiorrhiza inhibited MDA-MB-231 cell proliferation depending on cell line and point of time and concentration especially at 48h at high concentration.Salvia miltiorrhiza may enhance MDA-MB-231 cell caryocinesis via activing ERβpartly.It should be cautious to use Salvia miltiorrhiza to hormone insensitive breast cancer patients,the exact evidence need additional study in vivo.更多还原