黑豆乳清多肽抗皮肤光老化作用的研究

以昆明小鼠为对象,研究了黑豆乳清多肽对皮肤光老化的保护作用.试验设空白组(动物不照射紫外线),阳性对照组(脱毛区皮肤涂抹10％ Vc溶液),给药大、小剂量组(分别涂乳清多肽含量为15％和5％的乳液),模型组(涂抹蒸馏水)和基质组(涂抹基质乳液).除空白组外,其余各组小鼠均用长波紫外线照射12周.试验结束后处死动物,取受试部位皮肤进行切片观察和生化指标检测.结果表明,与模型组相比,给药大剂量组动物皮肤中CAT活力水平和HYP含量均显著升高(P＜0.01)、单位面积的皮肤质量和MDA含量显著降低(P＜0.01),皮肤红斑量和受损程度明显减轻.表明黑豆乳清多肽有抗皮肤光老化的作用.     

人参辅酶Q_(10)防晒霜对紫外线损伤小鼠的保护作用

目的研究人参辅酶Q_(10)防晒霜对紫外线损伤小鼠的保护作用。方法 36只小鼠随机分组,皮肤脱毛后分别涂抹相应霜剂,正常对照组不做任何处理,其他组进行紫外线照射,连续8周后,小鼠眼球取血处死进行指标检测。结果与正常组小鼠比较,模型组小鼠皮肤粗糙有皱纹、且角质层脱落,T-SOD与GSH-Px活力均明显下降,白细胞数与MDA含量明显增高(P0.05);与模型组比较,人参辅酶Q_(10)防晒霜组小鼠皮肤光滑、角质层没有脱落,T-SOD活力、红细胞与血红蛋白含量上升,白细胞数量、MDA含量与明显下降(P0.05),阳性对照组小鼠皮肤光滑、角质层没有脱落,GSH-Px活力上升,白细胞数量、MDA含量下降(P0.05)。结论人参辅酶Q_(10)防晒霜具有抗紫外线损伤的作用,其预防效果与甲氧基肉桂酸辛酯辅酶Q_(10)霜剂相当。     

五鹤续断总皂苷抗皮肤衰老的作用及其机制

目的:研究五鹤续断总皂苷抗皮肤衰老的作用及其机制。方法:将48只小鼠随机分为空白对照组、模型组、五鹤续断低、中、高剂量组和阳性对照组(n=8)。采用5%D-半乳糖(0.025 ml/(g·d))颈背部皮下注射制备皮肤衰老小鼠模型,五鹤续断低、中、高剂量组灌服五鹤续断总皂苷水溶液(50 ml/(kg·d)、100 ml/(kg·d)、200ml/(kg·d)),阳性对照组灌服维生素E(50 mg/(kg·d)),连续给药42 d后,检测各组小鼠皮肤组织羟脯氨酸(HYP)和脂褐质(LF)的含量,血清和皮肤组织过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)的活性与丙二醛(MDA)的含量。结果:与空白对照组比,模型组小鼠皮肤组织HYP含量明显减少,LF含量明显增多,血清和皮肤组织CAT、GSH-Px、SOD活性显著降低,MDA含量明显增加;与模型组比,五鹤续断低、中、高剂量和阳性对照组皮肤组织HYP含量明显增多,LF含量明显减少,血清和皮肤组织CAT、GSH-Px、SOD活性显著增强,MDA含量明显减少;与低剂量组比,高剂量和阳性对照组皮肤组织HYP含量明显增多,LF含量明显减少,血清和皮肤组织CAT、GSH-Px、SOD活性显著增强,MDA含量明显减少;血清和皮肤组织SOD活性与皮肤组织HYP呈显著正相关,与LF呈显著负相关。结论:五鹤续断总皂苷具有明显的抗皮肤衰老作用,其作用机制与抗氧化损伤密切相关。     

不同剂量的X-射线全身照射对小鼠健康状况及涎腺的影响

目的:通过直线加速器全身照射昆明小鼠建立辐射损伤模型,探索不同放射剂量对小鼠健康状况及涎腺功能和结构的影响。方法:选取八种不同剂量对昆明小鼠行体外全身照射,于照射后一个月内观察小鼠生长情况、体重变化;照射后一周、一个月检测各组小鼠血象的变化;测定放射半数致死剂量;照射后两个月,测定各组小鼠的唾液流量及唾液淀粉酶含量,并对下颌下腺组织切片行HE染色。结果:13Gy和15Gy照射组小鼠的体重逐渐下降,一周后死亡,其余组小鼠体重最终呈增加趋势。X-射线全身照射的半数致死量为10Gy。照射后一周,照射组小鼠的白细胞数目明显降低,与对照组比较有明显统计学差异(P0.01);在其他血象方面,除了7Gy组外,其他照射组与对照组比较也均有统计学差异(P0.05)。照射一个月后,各照射组小鼠的血象均恢复正常。照射后两个月,9Gy组和11Gy组小鼠的唾液流量及唾液淀粉酶含量均显著低于0Gy组,且11Gy组较9Gy组亦明显降低,差异均有统计学意义(P0.05)。随照射剂量的增加,小鼠的下颌下腺腺泡细胞数目逐步减少,结构排列紊乱,组织损伤逐渐加重。结论:X-射线全身照射引起小鼠健康状况受损,免疫功能减低,损伤程度与放射线强度呈剂量依赖性,小鼠半数致死量为10Gy,该剂量适合建立全身放射损伤模型。     

地鳖多肽提取物的抗氧化衰老机制

目的探讨地鳖多肽（ESW polypeptide）提取物抗氧化衰老机制的研究。方法小鼠连续腹腔注射D-半乳糖20 d,建立衰老模型,同时小鼠灌服不同剂量地鳖多肽提取物每日（0、40、80、160 mg/kg）,观察小鼠的正常活动、运动和耐应激能力。分别采用黄嘌呤氧化酶法、分光光度法检测小鼠血液和不同组织中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH-PX）活力,以及丙二醛（MDA）和还原型谷胱甘肽（GSH）含量;免疫荧光法检测细胞核转录因子2（Nrf2）在Caco-2细胞的表达。结果与对照和多肽组比较,衰老组小鼠体重增重缓慢、组织脏器系数降低、运动时间缩短、抗应激能力降低、组织中抗氧化酶活力降低。随着地鳖多肽剂量增加,多肽组小鼠体重增加明显,肝脏、脾脏和肾脏指数增加显著,小鼠静力和动力运动时间明显延长,小鼠耐缺氧、耐高温和运动时间延长并接近对照组,血液和不同组织中的SOD、CAT、GSH-PX活力及GSH含量显著提高,但MDA含量降低。与对照组比较,地鳖多肽组Caco-2细胞核内Nrf2表达量明显增加,接近阳性对照组。结论地鳖多肽可能通过启动Nrf2-ARE抗氧化信号通路,提高D-半乳糖致衰老小鼠抗应激和抗氧化能力,从而延缓小鼠氧化衰老。     

红花水提取液对小鼠系统性硬皮病的防治作用及其机制*

目的:研究红花水提取液对系统性硬皮病(SSc)模型小鼠的防治作用及相关机制研究。方法:60只 BALB /C小鼠随机分为对照组、模型组、强的松组、红花低、中、高剂量组,每组10只。对照组背部注射生理盐水,其余5组均背部皮下注射100 μl浓度为 200 μg /ml的注射用盐酸博来霉素,每天1次,连续注射28 d,制备SSc模型;造模同时对照组和模型组给予生理盐水10 ml/kg灌胃,强的松组给予强的松溶液4.5 mg/kg (10 ml/kg)灌胃,红花低、中、高剂量组分别给予红花1.5、3、6 g/kg (10 ml/kg)灌胃,各组均连续灌胃28 d。给药28 d后,取各组小鼠背部注射博来霉素区皮肤组织切片测量真皮厚度,采用水解法检测皮肤组织羟脯氨酸(HYP)含量;采用ELISA法检测皮肤组织结缔组织生长因子(CTGF)、转化生长因子-β(TGF-β)含量及血清白细胞介素-6(IL-6)、白细胞介素-17(IL-17)水平。结果:与对照组比较,模型组皮肤真皮厚度,皮肤组织CTGF、TGF-β、HYP含量及血清 IL-6、IL-17 水平明显升高(P＜0.05);与模型组比较,强的松组、红花低、中、高剂量组皮肤真皮厚度,皮肤组织 CTGF、TGF-β、HYP含量及血清 IL-6、IL-17水平明显降低(P＜0.05)。结论:红花水提取液可改善SSc小鼠皮肤状况(或真皮厚度),其作用机制可能与减轻免疫炎症反应有关。     

小分子金枪鱼多肽对小鼠记忆力、皮肤弹性及睡眠的影响

探究不同剂量的小分子金枪鱼多肽对小鼠记忆力、皮肤弹性和睡眠质量的影响。将6周龄的昆明小鼠随机分成4组(n=12),剂量组分别灌胃0.25 g/kg bw、0.5 g/kg bw和1.0 g/kg bw的小分子金枪鱼多肽,对照组灌胃PBS,连续6周。采用分光光度法分别测定小鼠脑组织SOD、GSH-Px和MDA含量,采用HE染色法制作小鼠皮肤石蜡切片,采用实时荧光定量PCR检测与记忆力、皮肤弹性和睡眠相关的基因表达。研究结果显示,小分子金枪鱼多肽能通过增加小鼠脑组织SOD和GSH-Px的含量,降低MDA的含量,上调记忆力相关基因的表达量来增强小鼠的记忆力;通过改善小鼠皮肤胶原纤维形态数目,改变皮肤弹性相关基因表达量来增加小鼠皮肤弹性;通过改变睡眠相关基因表达量来改善小鼠睡眠质量,且主要作用于GABAA通路和Hypocretin通路来发挥促眠作用。     

小鼠低剂量辐射损伤模型的初步研究

目的:对小鼠低剂量辐射损伤模型进行初步研究并筛选敏感检测指标.方法:实验分7组,1组为正常组,其余6组用XHA600直线加速器射线照射,每组取一部分小鼠于照射后2、4、8、16h测定红细胞(RBC)、白细胞(WBC)、血小板(PLT)和血红蛋白(HGB)含量;测定小鼠肝脏和脾脏重量及其组织中的超氧化合物歧化酶(SOD)和丙二醛(MDA)含量;剩余小鼠于4周后观察骨髓切片中血细胞变化.结果:照射后4h测定血常规,发现累积辐射0.4Gy组小鼠RBC有降低趋势,WBC、HGB显著降低,PLT显著升高;肝脏、脾脏的湿质量显著降低;小鼠肝脏组织中MDA的含量显著升高、SOD的活力显著降低;4周后小鼠骨髓细胞的病理改变与单次照射剂量相关,单次较大剂量(如0.6Gy)照射对骨髓细胞影响较大,在4周观察期不能自身恢复,而多次累积照射对骨髓细胞病理改变较小.结论:小鼠低剂量辐射损伤模型的最佳造模剂量为累积照射0.4Gy即每次照射100mGy,间隔一天,连续照射4次.     

维生素C对光化学法诱导皮肤光损伤的光保护作用:OCT定量研究

8-甲氧补骨脂素(8-MOP)联合UVA辐射(即光化学疗法PUVA)因诱导皮肤光损伤的副作用,常被用于实验动物皮肤光损伤模型的构建。为研究维生素C(Vc)对皮肤光损伤的保护效应,本研究在Balb/c小鼠背部皮肤涂抹0.1%8-MOP溶液1h后进行UVA辐照(10 J/cn2)。然后分别涂抹7.5%、15%和30%浓度的Vc溶液的进行光保护治疗,利用光学相干层析成像分析皮肤厚度和光信号衰减的变化,从而评估Vc对皮肤的光保护作用。结果显示,相对于溶媒组,Vc治疗组皮肤厚度减小,光衰减系数增加,其中15%效果尤为明显结果表明,局部涂抹Vc溶液具有一定的光损伤保护效应。     

灵芝提取液延缓皮肤衰老的实验研究

目的观察灵芝提取液延缓D-半乳糖致亚急性衰老小鼠皮肤衰老的作用。方法以D-半乳糖致亚急性衰老小鼠为模型,同时背部涂抹灵芝提取液,治疗42 d后,测定小鼠背部皮肤组织匀浆中超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量、脂褐质(LF)含量、羟脯氨酸含量。结果灵芝提取液低、中、高剂量均能不同程度提高皮肤中SOD活力,降低MDA含量,减少LF含量,提高小鼠皮肤中羟脯氨酸的含量(P0.01,P0.05),且具有明显的量效关系。结论灵芝提取液具有延缓D-半乳糖所致衰老小鼠的皮肤衰老作用。     

Effects of vitamin C deficiency on the skin of the senescence marker protein-30 (SMP30) knockout mouse

Senescence marker protein-30 (SMP30) is a gluconolactonase required for vitamin C (VC) synthesis. We examined effects of VC deficiency on the mouse skin using SMP30 knockout (KO) mice. SMP30 KO or wild type male mice were weaned around day 30 of age, and fed VC-deficient diet. They were given either VC water or control water. VC deficiency for 36 days did not affect skin hydroxyproline contents, while VC deficiency for 60 days decreased the hydroxyproline levels. Levels of some collagen mRNAs were different among the groups, but did not correlate with skin VC levels. The epidermis was morphologically abnormal in VC-deficient SMP30 KO mouse at 60 days after the weaning. Interestingly, the hair cycle was not synchronized among the groups. These data suggest low susceptibility of the mouse skin to VC deficiency and involvement of VC in the regulation of keratinocyte function and hair cycle in vivo.     

Effect of dietary histidine content on the change in content of skin urocanic acid isomers in hairless mice irradiated with ultraviolet B

Hairless mice were fed with a 10% amino acid mixture diet (control diet, 0.42% histidine content), the control diet without histidine (histidine-free diet), or the control diet rich in histidine (histidine-rich diet; histidine content, 4.2%) for 32 days. They were irradiated with UV light of 312 nm for 30 min, and skin samples were periodically taken for measuring the urocanic acid isomers. Total urocanic acid isomers were decreased by UV irradiation in all the three groups, the recovery being the fastest in the histidine-rich group. The percentage increase in cis-urocanic acid/total urocanic acid was quickly increased by UVB irradiation. The recovery of the ratio was slightly higher in the histidine-rich group, although the total urocanic acid level was higher in the histidine-rich group than in the others. Therefore, the absolute cis-urocanic acid content in the skin was almost the same among the three groups. These results indicate that the increased histidine intake strengthened UVB protection without any decrease in immune suppression.     

Wnt/β-catenin信号通路在博来霉素诱导的系统性硬化症小鼠模型血管重塑中的作用

系统性硬化症(systemic sclerosis,SSc)是一种慢性可累及全身多脏器的自身免疫性疾病,以广泛的血管病变及皮肤和内脏的纤维化为特征,但其机制迄今尚不明确。已有研究证实,Wnt通路参与了SSc纤维化,但其在血管病变中的病理作用尚未见报道。本研究拟采用博来霉素(bleomycin,BLM)诱导的SSc小鼠模型,探讨Wnt通路在SSc皮肤血管病变中的作用。将18只Balb/C小鼠随机平均分为3组,分别设为对照组(于小鼠背部皮下注射PBS 100 μL/d)、模型组(于小鼠背部皮下注射浓度为 1 mg/mL 博来霉素BLM 100 μL/d)和治疗组(于小鼠背部皮下注射 1 mg/mL BLM 100 μL/d,同时腹腔注射Wnt及β-catenin的抑制剂 iCRT3 5 mg/kg·d),于造模第28 d处死小鼠。小鼠皮肤取材后,通过HE染色及Masson染色观察到经BLM诱导的模型组小鼠背真皮、表皮厚度较对照组皮肤均明显增加(P<0.05),同时模型组的皮脂腺、毛囊等皮肤附属器明显减少,脂肪层厚度变薄并被纤维组织包绕,模型组皮肤胶原沉积较对照组增加;通过免疫组织化学染色在组织学层面鉴定α-SMA表达情况,发现模型组及治疗组α-SMA在皮肤组织中均高表达,α-SMA阳性表达在血管周围较对照组明显增加;通过ELISA方法检测出模型组小鼠血清中IL-6及IL-17表达量较对照组明显升高(P<0.05),治疗组小鼠血清中IL-6及IL-17的表达量较模型组明显下降(P<0.05);提取皮肤微血管片段,通过q-PCR检测到模型组及治疗组小鼠皮肤微血管中β-联蛋白的mRNA基因表达水平较正常组升高;通过Western印迹检测皮肤微血管Wnt5A、β-联蛋白、α-SMA、col1A1的蛋白质表达情况,发现纤维化相关蛋白质α-SMA及col1A1在模型组表达升高,较对照组有统计学差异(P<0.05),治疗组较模型组表达下降(P<0.05),Wnt通路相关蛋白质β-联蛋白及Wnt5A在模型组表达明显升高,较之对照组有统计学差异(P<0.05)。本研究提示,BLM能成功诱导小鼠系统性硬化症皮肤表型,Wnt通路的异常激活参与了BLM诱导的硬皮病小鼠皮肤微血管病变,特异性Wnt通路抑制剂iCRT3可能通过直接或间接的方式下调细胞因子IL-6及IL-17,从而降低BLM诱导的小鼠皮肤微血管中的α-SMA及col1A1蛋白质表达,改善小鼠皮肤微血管病变,干预BLM诱导的小鼠血管病变的进展。     

上海交通大学医学院附属新华医院崇明分院检验科

目的:探讨富血小板血浆(platelet-rich plasma,PRP)结合组织工程皮肤对裸鼠巨大创面修复的影响。方法:首先,利用密度梯度离心法制备富含生长因子的浓缩血小板的血浆,并测其所含生长因子的量;其次,在裸鼠的背侧部分构建大面积皮肤创面,分别用人工真皮,组织工程皮肤,碱性成纤维细胞生长因子组织工程皮肤,表皮生长因子组织工程皮肤和PRP结合组织工程皮肤修复裸鼠巨大创面;最后,手术后不同时间间隔收集组织标本,采用HE染色,PAS染色和免疫组化等方法评估创面愈合情况。结果:PRP结合组织工程皮肤组创面修复愈合情况最好。     

595 nm脉冲染料激光非损伤嫩肤治疗对皮肤羟脯氨酸含量的影响

羟脯氨酸的含量是反映胶原纤维变化的敏感生化指标.本文利用昆明小鼠作为动物模型,研究不同能量的595 nm脉冲染料激光对皮肤羟脯氨酸含量的影响.20只昆明小鼠随机分为两组,1个实验组和1个对照组.三种能量的激光各照射5次,每次间隔3天,光斑有10%的重叠.照射后测皮肤羟脯氨酸含量的变化,SPSS进行数据分析. 8 J/cm2、10 J/cm2和12 J/cm2组羟脯氨酸含量分别比对照组增加39% 、50% 和58%(p<0.05),12 J/cm2能量组的效果最好,但他们之间没有统计学上的显著差异,表明595nm的脉冲染料激光照射能明显增加皮肤羟脯氨酸的含量,用于非损伤嫩肤治疗是有效的.     

EPS and SMP dynamics at different heights of a submerged anaerobic membrane bioreactor (SAMBR)

Membrane bioreactors (MBR) technology for wastewater offers many advantages over conventional technologies such as high effluent quality, less footprint and others. The main disadvantage of membrane bioreactors (MBR) is related to membrane fouling, which is mainly caused by extracellular polymeric substance (EPS) and soluble microbial products (SMP). This research studied EPS and SMP dynamics at different heights of a submerged anaerobic membrane bioreactor (SAMBR). The SAMBR was operated under two organic loading rates (OLR) (0.79 and 1.56 kg/m³ d) and was fed with synthetic wastewater with glucose as the carbon source. The results showed percentages of chemical oxygen demand (COD) removal above 95% and the highest COD removal rates were observed at the bottom of the reactor (>83%) for both OLR. The EPS showed a stratification with highest quantities in the supernatant. For the SMP the highest concentration was in the bottom of SAMBR where utilization predominated associated products whereas in the SAMBR supernatant predominated biomass associated products. The OLR change led to a significant increase in SMP accumulation but not in EPS. These facts showed that EPS and SMP dynamic in the SAMBR seemed to be mainly influenced by biological activity, total suspended solids concentration and substrate composition.     

基于OCT技术的鼠皮肤激光热损伤修复过程的监测

采用Er:YAG激光(波长为2 940 nm,能量密度为:2.5 J/cm2单光斑,扫描次数为4)照射活体小白鼠皮肤,利用光学相干层析成像(optical coherence tomography,OCT)技术在活体小鼠上观察其皮肤组织在激光作用之前及作用之后光热损伤修复的整个过程,得到了激光光热作用下引起损伤的皮肤组织在此过程中皮肤光学特性参数的变化情况,发现皮肤修复过程中光学参数有显著差异,并分析了这些差异引起的原因,以揭示激光美容中并发症主要因素。     

Soluble microbial products (SMP) in anaerobic chemostats

The production of soluble microbial products (SMP) in anaerobic systems was evaluated using chemostat reactors. Results from steady-state and tracer experiments with (14)C-glucose and (14)C-acetate showed that significant amounts of SMP were produced during the acidogenesis of glucose, but that SMP did not accumulate during methanogenesis from acetate. In addition, at a retention time of 40 days, SMP comprised almost all of the effluent COD from the glucose-fed chemostat. For shorter retention times, as low as 10 days, the SMP concentration remained almost constant, but its significance in the effluent COD was reduced due to the accumulation of intermediate volatile fatty acids. The results from a (14)C-tracer experiment in the glucose-fed chemostat were used to evaluate the importance of including SMP formation and degradation in kinetic modeling of the methanogenic chemostats. Three models were evaluated: a model without SMP production, a model with SMP production but no degradation, and a model with SMP production and degradation, The results of this kinetic analysis indicate that the model that includes SMP production and degradation was the only one able to adequately represent the fate of (14)C in the tracer experiment. The kinetic parameters were successfully used to predict steady-state concentrations of SMP and to characterize the formation and degradation characteristics of the SMP. (c) 1994 John Wiley & Sons, Inc.     

Senescence Marker Protein 30 Has a Cardio-Protective Role in Doxorubicin-Induced Cardiac Dysfunction

Background

Senescence marker protein 30 (SMP30), which was originally identified as an aging marker protein, is assumed to act as a novel anti-aging factor in the liver, lungs and brain. We hypothesized that SMP30 has cardio-protective function due to its anti-aging and anti-oxidant effects on doxorubicin (DOX)-induced cardiac dysfunction.

Methods and Results

SMP30 knockout (SMP30 KO) mice, SMP30 transgenic (SMP30 TG) mice with cardiac-specific overexpression of SMP30 gene and wild-type (WT) littermate mice at 12–14 weeks of age were given intra-peritoneal injection of DOX (20 mg/kg) or saline. Five days after DOX injection, echocardiography revealed that left ventricular ejection fraction was more severely reduced in the DOX-treated SMP30 KO mice than in the DOX-treated WT mice, but was preserved in the DOX-treated SMP30 TG mice. Generation of reactive oxygen species and oxidative DNA damage in the myocardium were greater in the DOX-treated SMP30 KO mice than in the DOX-treated WT mice, but much less in the SMP30 TG mice. The numbers of deoxynucleotidyltransferase-mediated dUTP nick end-labeling positive nuclei in the myocardium, apoptotic signaling pathways such as caspase-3 activity, Bax/Bcl-2 ratio and phosphorylation activity of c-Jun N-terminal kinase were increased in SMP30 KO mice and decreased in SMP30 TG mice compared with WT mice after DOX injection.

Conclusions

SMP30 has a cardio-protective role by anti-oxidative and anti-apoptotic effects in DOX-induced cardiotoxicity, and can be a new therapeutic target to prevent DOX-induced heart failure.