雷公藤红素抑制MGC - 823细胞增殖和迁移及诱导凋亡

目的 探索雷公藤红素对MGC - 823细胞的增殖，迁移及诱导凋亡的作用，并探讨其作用机制。方法 0、1、2、4、6、8、12 μM雷公藤红素作用于MGC - 823细胞，采用MTT法检测增殖比率。0、1、2和4 μM雷公藤红素作用于MGC - 823细胞0、24、48和72 h，显微镜下观察细胞形态，采用细胞划痕实验检测细胞迁移能力，流式细胞术检测细胞凋亡，western blot法检测细胞中凋亡相关蛋白Bcl - 2、Bax、Akt、p - Akt等的表达水平。结果 经雷公藤红素作用后，MGC - 823细胞的增殖率均下降（P＜0.05）；细胞数量减少，体积缩小，皱缩变圆，部分不再贴壁，细胞核缩小；细胞迁移率减小（P＜0.05）；早期凋亡率增加（P＜0.05）；MGC - 823细胞中Bax表达升高，Bcl - 2和p - Akt表达降低（P＜0.05）。结论 雷公藤红素对MGC - 823细胞的增殖和迁移有抑制作用，并诱导其凋亡。     

雷公藤红素对人肺癌A549细胞增殖活性及能量代谢的影响

目的 探究雷公藤红素对人肺癌细胞（A549）的增殖活性以及能量代谢的影响。方法 噻唑蓝(MTT)法和生长曲线检测雷公藤红素对细胞的增殖抑制情况;HE染色观察细胞形态变化;分光光度法测定糖酵解途径中己糖激酶（HK）、丙酮酸激酶（PK）、乳酸脱氢酶（LDH）活力和三羧酸循环中琥珀酸脱氢酶（SDH）活力及能量代谢终产物ATP的含量。结果 雷公藤红素对A549细胞有增殖抑制作用,浓度在2 μg?mL^-1时,对A549细胞的抑制率达到62.71%,远远大于阳性药组5-FU的抑制率;生长曲线结果显示,雷公藤红素作用第4天细胞浓度最低,第6天细胞抑制程度最明显;HE结果表明雷公藤红素处理48 h后,细胞形态变圆,体积缩小,核深染且颜色变深;试剂盒检测结果显示,雷公藤红素非常显著的降低乳酸脱氢酶酶活力（P<0.01）,显著降低琥珀酸脱氢酶活力和ATP含量均显著降低(P<0.05),但对己糖激酶和丙酮酸激酶无明显作用。结论 雷公藤红素能够显著降低人肺癌A549细胞中乳酸脱氢酶和琥珀酸脱氢酶的活力,干扰能量代谢,抑制能量ATP生成,达到抑制A549细胞增殖的作用。     

雷公藤红素通过抑制MMP-9/sFlt-1/TNF-α减轻子痫前期模型大鼠症状的作用研究

目的:探讨雷公藤红素通过抑制金属基质酶-9/可溶性FMS样酪氨酸激酶-1/肿瘤坏死因子α(MMP-9/sFlt-1/TNF-α)减轻子痫前期模型大鼠症状的作用机制。方法:72只怀孕未经产的雌性SD大鼠随机分为6组:正常对照组、模型组、硫酸镁组(30mg·kg^-1)、雷公藤红素低、中、高剂量组(10,20,40 mg·kg^-1),每组12只。除正常对照组外,其余各组采用N-硝基-L-精氨酸甲酯盐酸盐诱导子痫前期模型,建模成功后各给药组灌胃给予相应药物,正常对照组和模型组给予等体积生理盐水。于妊娠期(第9天)测定大鼠24 h尿蛋白、血压、尿素氮(BUN),实时荧光逆转录法(RT-PCR)及蛋白质免疫印迹法(West blot)测定胎盘组织MMP-9 mRNA和蛋白水平,酶联免疫吸附法(ELISA)法测定sFlt-1、白细胞介素-4(IL-4)、TNF-α蛋白表达水平。结果:与模型组比较,硫酸镁组、雷公藤红素各剂量组收缩压、舒张压、24 h尿蛋白、BUN水平、MMP-9 mRNA和蛋白、sFlt-1、IL-4、TNF-α蛋白表达水平明显降低(P<0.05),且雷公藤红素各组呈剂量相关性(P<0.05);与硫酸镁组相比,雷公藤红素低、中剂量组以上各指标均明显升高(P<0.05)。结论:雷公藤红素能明显降低子痫前期模型大鼠血压、尿蛋白、BUN水平,缓解子痫前期症状,其机制可能与雷公藤红素通过抑制胎盘组织中MMP-9 mRNA和蛋白表达水平进而降低sFlt-1、IL-4、TNF-α蛋白表达有关。     

Celastrol protects kidney against ischemia–reperfusion-induced injury in rats

Background

Ischemia–reperfusion (IR) causes various damages in renal tissues, which is exacerbated by hypoxia-induced excessive inflammation and deteriorates the prognosis of patients after kidney surgery. Celastrol is a potent inflammation inhibitor that has little toxicity. In this report, we investigated whether celastrol protects against IR-induced renal injury in rats.

Materials and methods

Renal IR injury was induced by occlusion of the bilateral renal pedicles for 45 min followed by reperfusion for 6 h. Celastrol or vehicle solution was intraperitoneally injected 30 min before renal ischemia, respectively. Renal histology, function, and pro-inflammatory cytokines and mediators were assessed. The effect of celastrol on nuclear translocation of nuclear factor kappa B (NF-κB) was also measured.

Results

Celastrol significantly suppressed elevation of the renal function markers and the lipid peroxidation level, alleviated renal tubular damage, and decreased the levels of tumor necrosis factor-α, interleukin-1β, and monocyte chemotactic protein-1 (MCP-1) messenger RNA in kidney caused by IR. Moreover, celastrol prevented IR-induced expression of pro-inflammatory mediators, which was associated with suppression of nuclear translocation of NF-κB subunit p65.

Conclusions

Celastrol ameliorated the acute kidney injury caused by IR, which was associated with inhibiting local NF-κB activation and inflammation. Our findings suggest that celastrol could be useful for preventing IR-induced renal injury.     

南蛇藤醇抑制宫颈癌细胞生物学行为的机制研究

目的　探讨南蛇藤醇抑制宫颈癌细胞增殖和侵袭的作用机制。 方法　将正常培养HeLa细胞分为对照组、南蛇藤醇低、中、高剂量组（终浓度分别为4 μmol/L、8 μmol/L和12 μmol/L）、阳性对照组（顺铂,终浓度为10 μmol/L）、LY294002组（LY294002,终浓度为20 μmol/L）和（南蛇藤醇+LY294002）组（南蛇藤醇和LY294002,终浓度分别为12 μmol/L和20 μmol/L）。MTT法检测各组细胞存活情况;流式细胞仪检测细胞凋亡情况;Transwell检测细胞的侵袭;蛋白质印迹法检测PI3K/AKT/NF-κB信号通路的表达;采用皮下注射HeLa细胞建立裸鼠模型,观察南蛇藤醇（120 mg/kg）对裸鼠移植瘤体积和重量的影响。 结果　与对照组相比,南蛇藤醇组、阳性对照组和LY294002组HeLa细胞的相对增殖率、侵袭细胞数均明显下降（P<0.001）,细胞凋亡率明显升高（P<0.001）,p-PI3K、p-AKT、NF-κB p65蛋白表达明显下调（P<0.001）,且随着南蛇藤醇浓度升高,其作用增强;与LY294002组相比,（南蛇藤醇+LY294002）组HeLa细胞的相对增殖率和侵袭细胞数明显下降（q=10.182,q=10.217,P均<0.001）,细胞凋亡率明显升高（q=23.636,P<0.001）;与对照组相比,南蛇藤醇组裸鼠体重、移植瘤的体积和重量明显下降（q=4.100,P<0.020;q=13.501,P<0.001;q=5.078,P=0.005）。 结论　南蛇藤醇可能通过抑制HeLa细胞的PI3K/Akt/NF-κB信号通路,抑制其恶性生物学行为。     

雷公藤单体体外抑制胶质瘤细胞的实验研究

背景与目的：已有研究发现雷公藤具有抗肿瘤作用,本研究旨在探讨雷公藤单体对胶质瘤细胞的体外抑制作用。方法：通过MTT法测定3种雷公藤单体（甲素,红素和WilforolA)对胶质瘤细胞株SHG44,C6,U251的体外抑制作用;应用免疫组化法观察雷公藤甲素与雷公藤红素对SHG44胶质瘤细胞中Bax,Bcl-2蛋白表达的影响。结果：雷公藤二萜类单体雷公藤甲素对胶质瘤细胞有极显著的抑制作用;雷公藤三萜类单体中红素的抑制作用次之,两者均使SHG44细胞Bax表达增加,Bcl-2表达下降。结论：雷公藤甲素与雷公藤红素对胶质瘤细胞有明显的抗肿瘤作用,其作用与促进Bax表达,抑制Bcl-2表达,导致细胞凋亡有关。     

Celastrol regulates innate immunity response via NF-κB and Hsp70 in human retinal pigment epithelial cells

Elevated nuclear factor kappa B (NF-κB) activity and interleukin-6 (IL-6) secretion participates in the pathology of several age and inflammatory-related diseases, including age-related macular degeneration (AMD), in which retinal pigment epithelial cells are the key target. Recent findings reveal that heat shock protein 70 (Hsp70) may affect regulation of NF-κB. In the current study, effects of Hsp70 expression on NF-κB RelA/p65 activity were evaluated in human retinal pigment epithelial cells (ARPE-19) by using celastrol, a novel anti-inflammatory compound. Anti-inflammatory properties of celastrol were determined by measuring expression levels of IL-6 and endogenous NF-κB levels during lipopolysaccharide (LPS) exposure by using enzyme-linked immunosorbent assays (ELISA). Cell viability was measured by MTT and LDH assay, and Hsp70 expression levels were analyzed by Western blotting. ARPE-19 cells were transfected with hsp70 small interfering RNA (siRNA) in order to attenuate Hsp70 expression and activity of NF-κB RelA/p65 was measured using NF-κB consensus bound ELISA.Simultaneous exposures to LPS and celastrol reduced IL-6 expression levels as well as activity of phosphorylated NF-κB at serine 536 (Ser536) in ARPE-19 cells when compared to LPS exposure alone. In addition, inhibition of NF-κB RelA/p65 activity by celastrol was attenuated when Hsp70 response was silenced by siRNA. Favorable anti-inflammatory concentrations of celastrol showed no signs of cytotoxic response. Our findings reveal that celastrol is a novel plant compound which suppresses innate immunity response in human retinal pigment epithelial cells via NF-κB and Hsp70 regulation, and that Hsp70 is a critical regulator of NF-κB.     

雷公藤红素体外抑制人类急性髓系白血病细胞的实验研究

 目的　研究雷公藤红素体外对人类急性髓系白血病（AML）细胞的体外抑制作用。方法　应用四甲基偶氮唑蓝（MTT）法和流式细胞术（FCM）等方法研究雷公藤红素对人类AML细胞的影响。结果　MTT实验显示,与空白组比较,培养不同时间后不同浓度组雷公藤红素的细胞增殖抑制率均明显升高,差异有统计学意义（P＜0.01）。FCM检测结果显示,1 μmol／L以上浓度雷公藤红素作用不同时间,AML细胞均可出现凋亡现象,凋亡率明显高于不加药物的对照组（P＜0.01）。结论　雷公藤红素对AML有明显的抑制作用,其作用可能与其诱导细胞凋亡有关。     

南蛇藤素抑制豚鼠体外精子的受精能力

南蛇藤素(Cel)对豚鼠精子前向运动(FM)、获能(Cap)、顶体反应(AR)和穿透去透明带仓鼠卵(SPA)均有明显的抑制作用,其作用强度随剂量而增加;对Cel的敏感性依次为精子Cap>FM>SPA>AR。Cel对豚鼠精子AR,FM和Cap的抑制作用比乙酸棉酚(GA)明显为强。