参地补肾胶囊含药血清对高糖诱导转分化HK-2细胞ZO-1、α-SMA表达的影响

目的:通过观察参地补肾胶囊含药血清对高糖诱导转分化的人肾小管上皮(HK-2)细胞ZO-1(紧密连接蛋白)、α-SMA(α平滑肌肌动蛋白)表达的影响,从分子生物学水平探讨参地补肾胶囊干预肾小管上皮细胞转分化的机制。方法:制备空白及参地补肾胶囊、贝那普利含药血清,以高糖诱导的人肾小管上皮细胞(HK-2)作为观察对象,实验分为正常对照组、高糖模型组、参地补肾胶囊组和贝那普利组,各组HK-2细胞培养48 h,采用Western Blot方法检测ZO-1、α-SMA蛋白的表达情况,Real-time PCR技术检测ZO-1mRNA的表达情况。结果:与高糖模型组比较,贝那普利组、参地补肾胶囊组HK-2细胞ZO-1表达量增加(P 0. 05),参地补肾胶囊组ZO-1表达较贝那普利组增强(P 0. 05);贝那普利组及参地补肾胶囊组HK-2细胞a-SMA表达均低于高糖模型组(P 0. 05),参地补肾胶囊组aSMA表达低于贝那普利组(P 0. 05);参地补肾胶囊组ZO-1mRNA表达增加(P 0. 05),高于贝那普利组(P 0. 05)。结论:参地补肾胶囊可能是通过调节细胞ZO-1、α-SMA的表达抑制肾小管上皮细胞向肌成纤维细胞的转分化。     

当归红芪超滤物对放射性H9C2细胞损伤TNF-α/CTRP9信号通路相关因子表达的影响

目的观察当归红芪超滤物对放射性H9C2细胞损伤的影响,探讨其对心肌损伤的保护作用机制。方法采用大鼠灌胃方法制备药物血清。将H9C2细胞随机分为空白组、阴性对照组、模型组、盐酸贝那普利组、当归红芪超滤物组和盐酸贝那普利+当归红芪超滤物组。药物血清干预及X线照射H9C2细胞后,ELISA检测肌钙蛋白T(cTnT)、肌钙蛋白Ⅰ(c TnⅠ)及C-反应蛋白(CRP)含量,Western blot和RT-PCR检测肿瘤坏死因子-α(TNF-α)和肿瘤坏死因子相关蛋白9(CTRP9)的表达,免疫荧光检测转化生长因子-β(TGF-β)和α-平滑肌肌动蛋白(α-SMA)的表达。结果与空白组比较,模型组H9C2细胞c Tn T、c TnⅠ、CRP含量明显增加,TNF-α、CTRP9蛋白和基因表达明显升高,α-SMA和TGF-β蛋白表达明显升高;与模型组比较,盐酸贝那普利组、当归红芪超滤物组和盐酸贝那普利+当归红芪超滤物组cTnT、c TnⅠ、CRP含量明显减少,TNF-α、CTRP9蛋白和基因表达明显降低,α-SMA和TGF-β蛋白表达明显降低。结论当归红芪超滤物可通过下调TNF-α/CTRP9信号通路相关因子的表达,达到保护放射性H9C2细胞损伤的目的。     

肾炎2号对IgA肾病大鼠α-平滑肌肌动蛋白表达的实验研究

目的:探讨肾炎2号对IgA肾病大鼠肾小球内α-平滑肌肌动蛋白(α-SMA)表达的影响。方法:使用口服和尾静脉注射牛血清白蛋白(BSA)的方法进行IgA肾病模型的制作,治疗组、对照组造模同时分别给予肾炎2号(15g/kg)和福辛普利(4.17mg/kg)至12周末。观察尿常规、24h尿蛋白定量、肾组织的病理改变及肾小球内α-平滑肌肌动蛋白(α-SMA)的表达。结果:①第7~12周末模型组的24h尿蛋白定量明显高于正常组、治疗组与对照组(P<0.01)。②与模型组比较,治疗组大鼠肾小球内α-SMA的表达减弱(P<0.01),但治疗组、对照组相比无明显统计学上的差异(P>0.05)。结论:肾炎2号具有减轻肾小球内α-SMA的表达的作用。     

肾疏宁防治肾小球硬化机制的实验研究

目的:观察肾疏宁防治肾小球硬化的作用机制。方法:采用单侧肾切除并阿霉素尾静脉注射建立大鼠肾小球硬化模型,观察肾疏宁给药8周后肾小球病理表现,免疫组织化学染色法观察肾小球内α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)、层粘连蛋白(LN)、Ⅳ型胶原(ColⅣ)阳性表达,并进行图像分析。结果:肾疏宁治疗组肾小球内α-SMA、FN、LN、ColⅣ和系膜基质较模型组显著减少,与苯那普利无显著性差异。结论:肾疏宁能抑制肾小球系膜细胞表型转化,减少细胞外基质蓄积,对阿霉素肾小球硬化大鼠肾脏具有保护作用。     

肾病Ⅰ号方对单侧输尿管结扎大鼠肾间质纤维化的影响

目的:观察肾病Ⅰ号方对肾纤维化大鼠α平滑肌肌动蛋白(α-SMA)、Ⅳ型胶原(COL-Ⅳ)和转化生长因子-β1(TGF-β1)表达的影响。方法:24只雄性SD大鼠随机分为假手术组、UUO模型组、洛丁新组和肾病Ⅰ号方组,每组6只。采用单侧输尿管结扎的方法制备大鼠肾纤维化模型。假手术组和UUO模型组每日予2 m L生理盐水灌胃,洛丁新组按洛丁新片1.67 mg·kg~(-1)·d~(-1)给药,肾病Ⅰ号方组按18.75 g·kg~(-1)·d~(-1)给药。灌胃14 d后处死大鼠,采集血清检测尿素氮(blood urea nitrogen,BUN)和血肌酐(serum creatinine,SCr);取肾组织行HE染色评价肾小管间质纤维化损伤程度;采用免疫组织化学染色方法检测α-SMA、COL-Ⅳ、TGF-β1在肾组织中的表达情况;免疫印迹试验(Western blot)检测α-SMA相对蛋白表达量。结果:与模型组比较,洛丁新组、肾病Ⅰ号方组大鼠的SCr,BUN明显下降(P<0.05),肾组织α-SMA、COL-Ⅳ、TGF-β1表达量下降显著,差异具有统计学意义(P<0.05)。结论:肾病Ⅰ号方对肾纤维化具有一定改善作用,其作用机制可能是通过下调α-SMA、COL-Ⅳ、TGF-β1的表达,从而达到肾脏保护的作用。     

雷公藤多苷对不可逆性肾小球硬化模型系膜损伤的抑制作用

目的观察雷公藤多苷(GTW)对不可逆性肾小球硬化模型系膜损伤的抑制作用,阐明其在体内延缓肾小球硬化的分子药理机制。方法运用2次注射单克隆抗体(mAb)1223的方法,建立大鼠不可逆性肾小球硬化模型,以经口灌胃GTW(50mg·kg^-1BW)干预42d,并设立对照组。比较24h尿蛋白排泄量(Upro)、肾功能(Scr,BUN)以及肾小球形态学(LM,IF)变化,并检测(RTPCR)模型鼠肾组织中I型胶原(collagentypeI)、转化生长因子(TGFβ)mRNA表达水平。结果GTW抑制肾小球硬化模型鼠Upro和系膜增殖。肾小球总细胞数GTW组(65.67±3.43),对照组(87.02±2.41),P<0.05;细胞外基质积分GTW组(1.20±0.06),对照组(2.77±0.23),P<0.05;α平滑肌肌动蛋白积分GTW组(1.75±0.33),对照组(2.62±0.15),P<0.05;I型胶原积分GTW组(1.68±0.31),对照组(2.06±0.24),P<0.05。GTW使肾小球硬化模型鼠肾组织中collagentypeI,TGFβmRNA表达水平下调53.5%和14.7%,与对照组比较,二者差异均有显著性(P<0.05)。结论GTW抑制肾小球硬化模型鼠Upro和系膜增殖的作用是通过减少细胞因子mRNA的表达来实现的。     

IgA肾病中肾小球系膜细胞表型转化及肾络宁的防治作用

目的探讨IgA肾病大鼠肾小球系膜细胞的表型转化及中药复方肾络宁的作用。方法SD大鼠随机分为正常组、模型组、肾络宁组、肾炎康复片组;采用牛血清白蛋白和葡萄球菌肠毒素B复合感染的方法建立大鼠IgA肾病模型;常规检测尿红细胞、尿蛋白、血肌酐,光镜、电镜、荧光免疫观察肾组织病理;免疫组织化学染色和计算机图象分析系统观察大鼠肾小球系膜细胞α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)、细胞间黏附分子(ICAM-1)的表达情况。结果模型组肾小球系膜区α-SMA、FN、ICAM-1明显阳性表达(P<0.01),系膜区重度IgA沉积,系膜细胞中度增生,基质增多,毛细血管狭窄,血尿及蛋白尿、血肌酐明显增高;肾络宁及肾炎康复片组α-SMA、FN、ICAM-1表达明显减低(P<0.01),IgA轻度沉积,系膜细胞及基质轻度增多,血尿、蛋白尿及血肌酐明显减低。结论IgA肾病大鼠肾小球系膜细胞发生表型转化,中药复方肾络宁可明显抑制IgA肾病大鼠肾小球系膜细胞的表型转化,改善临床及病理变化,肾络宁是防治IgA肾病的有效方药。     

贝那普利联合参麦注射液治疗糖尿病肾病的临床观察

目的探讨血管紧张素转换酶抑制剂(贝那普利)联合参麦注射液治疗糖尿病肾病的临床疗效。方法将68例糖尿病肾病患者随机分为治疗组与对照组,两组均给予基本降糖治疗。治疗组给予贝那普利合参麦注射液,对照组仅给予贝那普利。结果治疗组疗效优于对照组,治疗组24 h尿蛋白定量(24 hUAE),尿素氮(BUN),血肌酐(Cr)等指标的改善优于对照组。结论贝那普利联合参麦注射液治疗糖尿病肾病的疗效较单用贝那普利为佳。     

益肾通络方对膜性肾病大鼠足细胞表型转化的影响

目的通过观察膜性肾病(MN)足细胞标志蛋白Nephrin、α-平滑肌肌动蛋白(α-SMA)的变化,探讨益肾通络方对膜性肾病大鼠足细胞转分化的干预作用及其可能机制。方法从60只实验鼠中随机选取15只为正常对照组,其余通过尾静脉注射阳离子化牛血清白蛋白(C-BSA)法建立膜性肾病动物模型,造模成功后将大鼠随机分为模型组、贝那普利(10 mg/kg)组、益肾通络方(21.33 g/kg)组。各组依据对应的剂量分别灌胃给药,治疗结束后检测24 h蛋白尿(UTP)、甘油三酯(TG)、总胆固醇(TC)、白蛋白(ALB)、总蛋白(TP)、肌酐(Scr)、尿素氮(BUN)评估大鼠治疗情况;观察大鼠的肾脏病理形态学变化;免疫组化检测大鼠肾组织Nephrin、α-SMA蛋白表达。结果经贝那普利和益肾通络方干预后,UTP、TC、TG较模型组显著下降,TP、ALB较模型组显著增高(P0.05);BUN、Scr差异无统计学意义(P0.05)。免疫组化显示,贝那普利组与益肾通络方组Nephrin表达较模型组上升,a-SMA表达较模型组降低。结论益肾通络方能够降低膜性肾病大鼠蛋白尿,升高血浆白蛋白,降低血脂,其作用机制可能与抑制足细胞发生表型转分化有关。     

肾衰泄浊汤及其补虚、祛邪组分对UUO大鼠肾间质纤维化的影响

目的:研究肾衰泄浊汤及其补虚,祛邪组分对单侧输尿管梗阻(UUO)大鼠肾间质纤维化的影响。方法:通过结扎单侧输尿管建立UUO大鼠模型。SD大鼠随机分为6组,分别为假手术组,模型组,贝那普利组,肾衰泄浊汤组,补虚方组,袪邪方组;每组24只。手术后第3天,肾衰泄浊汤组,补虚方组,袪邪方组大鼠均按0. 8 g·mL-1肾衰泄浊汤浓缩剂给药,即给药剂量为8. 0 g·kg-1·d-1体质量灌胃;贝那普利组大鼠给贝那普利1. 5 mg·kg-1·d-1体质量灌胃,假手术组,模型组大鼠给等量生理盐水灌胃。术后第7,14,21天,分别用蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应(Real-time PCR)检测肾组织中周细胞及相关信号通路标志物表达情况。结果:与模型组比较,肾衰泄浊汤组,补虚方组,袪邪方组在UUO后第14,21天的间质损伤评分和间质胶原的积累均显著降低(P0. 05)。除假手术组外,各组在第14,21天的肾组织中α-平滑肌肌动蛋白(α-SMA),血小板衍生生长因子A(PDGFA),神经/胶质细胞2型硫酸软骨素糖蛋白(NG-2),血管内皮生长因子A(VEGFA),血管内皮生长因子受体1(VEGFR1),血小板源性生长因子β(PDGF-β),血小板源性生长因子受体β(PDGFR-β)的蛋白表达均较第7天显著增加(P0. 05)。在手术后第21天,肾衰泄浊汤组的肾组织中NG-2,VEGFA,VEGFR1,PDGFR-β蛋白表达均显著低于模型组(P0. 05)。与模型组比较,肾衰泄浊汤组在手术后第21天的肾组织中α-SMA,PDGFA,NG-2,VEGFA,VEGFR1,PDGF-β,PDGFR-βmRNA相对表达量均显著降低(P0. 05),并且肾衰泄浊汤组在手术后第21天α-SMA,NG-2,VEGFA,VEGFR1,PDGFR-βmRNA相对表达量显著低于贝那普利组(P0. 05)。结论:肾衰泄浊汤复方及其补虚,祛邪组分对UUO大鼠肾间质纤维化均具有拮抗作用,其作用机制与抑制周细胞及其相关细胞信号通路激活有关。     

半夏、掌叶半夏及禹白附凝集素蛋白的刺激性研究

目的:研究半夏凝集素(Pinellia ternata agglutinin,PTA),掌叶半夏凝集素(Pinellia pedtaisecta agglutinin,PPA),禹白附凝集素(Typhonium giganteum agglutinin,TGA)的刺激性作用。方法:SD雄性大鼠随机分为空白组,PTA,PPA及TGA高中低剂量组(300,200,100 mg.L-1),ip给药,4 h后处死大鼠,ip生理盐水,取腹腔液测中性粒细胞迁移数目,测定腹腔液中总蛋白,一氧化氮(NO),前列腺素E2(PGE2)的含量;家兔随机分为空白组,半夏,掌叶半夏及禹白附的凝集素组及毒针晶组,半夏,掌叶半夏及禹白附的针晶加其凝集素组,观察家兔眼结膜充血,水肿及分泌物,考察PTA,PPA,TGA的刺激性作用。结果:大鼠ip PTA,PPA,TGA后,与空白组腹腔液中性粒细胞迁移数目(633.33±233.81)×106/L比较,100 mg.L-1组PTA(1 083.33±116.9)×106/L,PPA(1 033.3±150.55)×106/L及TGA(1 133.33±103.28)×106/L均具极显著性差异(P<0.01),均可引起严重的中性粒细胞向腹腔迁移,且浓度升高,作用增强;大鼠腹腔液的总蛋白,NO及PGE2的含量显著升高,均可加重针晶刺激家兔眼结膜强烈水肿。结论:天南星科中具有刺激性毒性作用的半夏,掌叶半夏及禹白附所含凝集素蛋白具有刺激性作用,是其毒性刺激性成分。     

Scientific validation of the different purification steps involved in the preparation of an Indian Ayurvedic medicine, Lauha bhasma

Ethnopharmacological relevance

Lauha bhasma (iron ash) is one of the iron-based herbo-metallic preparations used in Ayurvedic medicine for treating various ailments due to iron deficiency.

Materials and methods

The preparation of Lauha bhasma (iron ash) requires normal purification (heat treatment in vegetable and animal products), special purification (treatment with herbal constituents) and calcination steps aimed at converting the raw material to a suitable therapeutic form. In this study, we have systematically and scientifically evaluated through a series of qualitative tests and modern analytical tools the importance of the treating media.

Results

Our data demonstrates that these steps are necessary to remove the grease and scales in the raw material. While heating, microcracks appeared on the surface of the iron, which improved the reactivity with the herbal constituents in addition to incorporating nanostructured features. Further, the use of plant products facilitated the removal of Fe³⁺ present in the raw material by forming soluble complexes. The Fe²⁺ present in the raw materials also forms an insoluble complex with the herbal constituents in the presence of UV radiation.

Conclusions

In conclusion, our data summarily suggest that the purification steps involved in the preparation of Lauha bhasma (iron ash) are critical.     

2株内生真菌对菊花抗旱特性的影响

目的:以PEG6000模拟干旱条件,研究接种内生真菌(葡萄孢属C1菌Botrytis sp.、球毛壳菌G4菌Chaetomium globosum对药用菊花Ch.morifolium抗旱性的影响.方法:分别用0%,10%,20%,30%,40%PEG6000胁迫菊花组培苗4 d,测定各处理组菊花生物量,叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)活性及叶片丙二醛(MDA)、可溶性蛋白含量.结果:模拟干旱胁迫后,接种内生真菌的菊花长势好于对照(未接菌),PEG6000胁迫4 d后,随着胁迫浓度的增加,菊花总生物量不断减少,接菌组生物量显著高于对照,C4组高于C1组;各处理组MDA含量随着胁迫浓度的增加而不断增加;各处理组POD活性、可溶性蛋白含量随着胁迫浓度的增加均呈现先增加后减小的趋势;对照SOD活性随着胁迫浓度的增加而逐渐增加,接菌组SOD活性基本保持不变;对照PAL活性随着胁迫浓度的增加而逐渐增加.不同浓度PEG胁迫后,接菌组MDA含量始终低于对照,SOD活性、POD活性、可溶性蛋白含量、PAL活性均始终高于对照.结论:接种内生真菌提高菊花的抗旱能力.     

Prophylactic and curative activities of extracts from Warburgia ugandensis Sprague (Canellaceae) and Zanthoxylum usambarense (Engl.) Kokwaro (Rutaceae) against Plasmodium knowlesi and Plasmodium berghei

Ethnopharmacological relevance

This work reports the anti-plasmodial activities of Warburgia ugandensis and Zanthoxylum usambarense commonly used as phytomedicines against malaria by some Kenyan communities.

Aim of study

To determine the anti-plasmodial activities of extracts from Warburgia ugandensis and Zanthoxylum usambarense against Plasmodium knowlesi and Plasmodium berghei.

Materials and methods

Eight plant extracts were screened for in vitro anti-plasmodial activity against Plasmodium knowlesi, in a 96-well plate incubated at 37 °C on a RPMI culture medium supplemented with baboon serum. Of the eight, three were investigated for prophylactic and curative activities in BALB/c mice against drug-sensitive Plasmodium berghei in a 4-day test at a dose rate of 200 mg/kg/day.

Results

Inhibitory concentrations (IC₅₀) values of between 3.14 and 75 μg/ml, up to 69% chemosuppression of parasites growth and over 80% survivorship of treated mice were observed.

Conclusion

The two medicinal plants, Warburgia ugandensis and Zanthoxylum usambarense possess bioactive compounds against malaria parasites and could be exploited for further development into malaria therapy.     

Citrus limetta leaves extract antagonizes the hypertensive effect of angiotensin II

Ethnopharmacological relevance

Citrus limetta Risso (Rutaceae) is widely used in Mexico for healing purposes, among them as antihypertensive treatment.

Aim of the study

To assess the antihypertensive effect of C. limetta leaves as one of its ethnomedical uses.

Materials and methods

The acute response of blood pressure to angiotensin II administration was measured in mice. Additionally, the acute oral toxicity profiles were determined.

Results

The findings of the current investigation showed that different concentrations of the aqueous extract prevented the raise of systolic blood pressure (p ≤ 0.001 vs. vehicle), diastolic blood pressure (p ≤ 0.0002 vs. vehicle) and mean blood pressure (p ≤ 0.0000 vs. vehicle); with a dose dependent effect for diastolic pressures at 125–500 mg/kg dosages. The 500 and 1000 mg/kg doses inhibited the action of Ang II in similar extent to telmisartan. Toxic signs or deaths were not observed in mice treated at 2000 mg/kg of C. limetta extract.

Conclusions

All doses of C. limetta aqueous extract, used in this assay, were safe and effective.     

Lipoxygenase inhibitory activity of crude bark extracts and isolated compounds from Commiphora berryi

Ethnopharmacological relevance

Commiphora berryi is traditionally used for the treatment of cold and fever as well as for wound healing in the southern parts of India.

Aim of study

The present study was designed to investigate in vitro soybean lipoxygenase inhibitory activity of crude extracts and compounds isolated from Commiphora berryi.

Materials and methods

The bark of Commiphora berryi was extracted with different organic solvents and subjected to chromatographic separation for isolation of bioactive compounds. Structures of isolated compounds were elucidated by spectroscopic methods. The anti-inflammatory activity of bark extracts and bioactive compounds were assessed by in vitro soybean lipoxygenase (SBL) assay.

Results

3β-Hydroxyglutin-5-ene (1), friedelin (2), cycloeucaneol (3) nimbiol (4), sugiol (5), surianol (6), daucosterol (7) and ursolic acid (8) were isolated from crude bark extracts of the Commiphora berryi. The structure of nimbiol (4) was also confirmed by single crystal X-ray analysis. The petroleum ether, methanol, chloroform and ethyl acetate extracts of bark of Commiphora berryi showed SBL inhibitory activity with the IC₅₀ values of 15.3, 54.2, 71.5 and 87.8 μg/ml respectively. Among all the isolates, friedelin (2) showed significant SBL inhibitory activity with IC₅₀ 35.8 μM.

Conclusion

The overall results provide evidence that the studied plant might be a potential source of anti-inflammatory agents.     

Anti-inflammatory compounds of “Qin-Jiao”, the roots of Gentiana dahurica (Gentianaceae)

Ethnopharmacological relevance

“Qin-Jiao” is a well-known traditional Chinese medicinal (TCM) herb having been used generally for fighting rheumatoid arthritis (RA) since ancient times. The root of Gentiana dahurica Fisch (Gentianaceae) is one of the four officially validated “Qin-Jiao” as listed in the Chinese Pharmacopoeia. In addition, it is a common Tibetan medicinal herb used for the treatment of tonsillitis, urticaria, and RA, while the flowers have been used as a Mongolian herb for curing cough sore throat and eliminating the phlegm due to its anti-inflammatory effect.

Aim of the study

The aim of the study was to characterize the anti-inflammatory compounds in “Qin-Jiao”, on the basis of detailed investigation on not only the phytochemical study of Gentiana dahurica, but also the bioactive evaluation on compounds obtained presently and previously from different “Qin-Jiao” origins and Gentiana species.

Materials and methods

The ethanol extract of air-dried roots of Gentiana dahurica was suspended into H₂O and extracted with EtOAc and n-BuOH, successively. Repeated column chromatography (CC) and semi-preparative HPLC were carried out on each of the fractions. The isolated compounds were determined by detailed spectroscopic analysis and acidic hydrolysis. Anti-inflammatory activities of 18 isolates, together with 12 typical compounds obtained previously by our group from the other “Qin-Jiao” origins (Gentiana crassicaulis, Gentiana straminea) and Gentiana rigescens, were tested by inhibitory effects on LPS-induced NO production in macrophage RAW264.7 cells and TPA-induced cyclooxygenases-2 and -1 (COXs-2/1) production on zebrafish model.

Results

A new lignan glycoside (1) was identified, together with 20 known compounds, including 10 iridoid glycosides (2–11), three steroids (12–14), four lignans (15–18), one phenylpropanoid (19) and two triterpenes (20–21). Anti-inflammatory bioassay showed that only compound 21 displayed potential inhibitory effect on NO production (IC₅₀=16.85 μM), while 20 tested compounds had inhibitory activities on COXs-2/1. Among them, the triterpenoid 21 was the most active compound with an inhibitory value of 78% at a concentration of 30 μM. All the tested compounds showed no cytotoxicity on five human cancer cell lines (40 μM) and zebrafish (30 μM), except for 21 displaying weak cytotoxicity on human myeloid leukemia HL-60 (IC₅₀=16.43 μM).

Conclusion

Most of compounds particularly iridoid glycosides from “Qin-Jiao” display potential inhibitory effect on COXs-2/1. The results support the historical importance of the well-known TCM herb, “Qin-Jiao”, having been commonly used for fighting RA. As major components, the bioactive iridoid glycosides should play important role in the anti-inflammatory effect of “Qin-Jiao”. Although further research will be required to evaluate the selective activities of the COXs-2/1 inhibitors, this work validates the medicinal use of “Qin-Jiao” and provides information for different “Qin-Jiao” origins having different treating effects on RA.     

Immunomodulation of RAW264.7 macrophages by GLIS, a proteopolysaccharide from Ganoderma lucidum

The immunomodulatory effect of Ganoderma lucidum immunomodulating substance (GLIS) on macrophages has been investigated as part of on-going research into the anti-cancer properties of Ganoderma lucidum. Proliferation of bone marrow macrophages (BMMs) was enhanced by GLIS in a dose-dependent manner. Microscopic examination revealed that numerous GLIS-treated RAW264.7 macrophages were enlarged and formed pseudopodia. Exposure of RAW264.7 macrophages to GLIS resulted in significant increases in NO production, induction of cellular respiratory burst activity, and increased levels of IL-1beta, IL-12p35 and IL-12p40 gene expression. Our data indicate that GLIS activates the immune system by modulating cytokine production.     

独活挥发油对N-脂肪酰基乙醇胺水解酶的抑制作用及抗炎作用研究

目的:研究独活挥发油对内源性大麻素水解酶N-脂肪酰基乙醇胺水解酶(N-acylethanolamine-hydrolyzing acid amidase,NAAA)水解活性的影响以及对脂多糖(LPS)诱导的小鼠巨噬细胞RAW264.7炎症反应模型的抗炎作用.方法:采用水蒸气蒸馏法提取独活挥发油,GC-MS检测化学成分;采用LC-MS检测NAAA水解活性;采用LPS诱导RAW264.7细胞建立细胞炎症反应模型;采用LC-MS检测细胞内棕榈酸乙醇胺(N-palmitoylethanolamine,PEA)水平;采用实时定量PCR检测肿瘤坏子因子-α(TNF-α)、诱导型一氧化氮合酶(iNOS)和白细胞介素-6(IL-6)mRNA表达;采用酶联免疫吸附法(ELISA)检测TNF-α含量;采用Griess法检测一氧化氮(NO)含量.结果:独活挥发油可抑制NAAA水解活性,升高LPS诱导的RAW264.7细胞内PEA水平;独活挥发油可下调LPS诱导的RAW264.7细胞炎症因子TNF-α,iNOS,IL-6 mRNA表达;独活挥发油可抑制LPS诱导的RAW264.7细胞TNF-α和NO释放.结论:独活挥发油能够抑制NAAA水解活性,升高细胞内PEA水平,降低炎症因子表达,具有一定的抗炎作用.     

Therapeutic effects of dichloromethane fraction of Securidaca inappendiculata on adjuvant-induced arthritis in rat

Ethnopharmacological relevance

Securidaca inappendiculata (SI) is a traditional antirheumatic medicine used in China. The present study was designed to investigate the therapeutic efficacy of dichloromethane fraction of SI (SID) at three different doses on adjuvant induced arthritis (AA) rats.

Methods

Arthritis severity was evaluated by arthritic score, body weight loss, paw circumference, histological changes and hyperplasia of lymphatic tissues. Serum samples were collected for estimation of superoxide dismutase (SOD), glutathione (GSH), hydroxy radical (OH·), nitric oxide (NO), malondialdehyde (MDA), N-acetyl glucosaminidase (NAG), sialic acid (SA), alkaline phosphatase (ALP), aspartate transaminase (AST) and alanine transaminase (ALT). The levels of GSH, MDA, NAG and SA in liver were also assessed. The levels of interleukin-1 (IL-1), tumor necrosis factor alpha (TNF-α), monocyte chemotactic protein 1 (MCP-1) and vascular endothelial growth factor (VEGF) were determined using ELISA method. Another portion of blood was used for total and differential leucocyte counts.

Results

Administration with SID (at high dose with 100 mg/kg) significantly ameliorated the AA severity, suggested by the modulatory effects on body weight loss, paw swelling, hyperplasia of lymphatic tissues and synovial membrane, neutrocytosis and lymphocytosis. It also decreased levels of NO, MDA and OH·, restored SOD and GSH levels in serum. The abnormal increased levels of AST, ALT, ALP, NAG and SA significantly were reverted (compared with AA rats, P<0.01). A similar result was observed in livers. Levels of IL-1, TNF-α, MCP-1 and VEGF were reduced dramatically by SID too.

Conclusion

The results suggest SID possesses substantial anti-arthritic activity. The therapeutic efficacy may be due to immumodepressive effects, cytokines regulation, increasing membrane stability and antioxidantive activity.