木犀草素对大鼠缺血性脑组织上调Nrf2、Bcl-2下调Bax的表达

目的探讨木犀草素对缺血性脑组织的保护机制。方法用线栓法制备大鼠大脑中动脉闭塞(MCAO)模型。实验采用Western blot和RT-q PCR观察Nrf2、Bcl2、Bax蛋白和基因表达变化,比较各组脑梗死体积、患侧脑水肿和神经功能缺损评分。结果木犀草素可显著改善神经功能缺失,减轻脑水肿,减小脑梗死体积(P<0.05)。木犀草素干预后,能明显诱导保护性基因Nrf2的表达,升高Bcl2的表达,降低Bax的表达(P>0.05)。结论 Nrf2、Bcl2和Bax可能是木犀草素的作用靶点,木犀草素可能是通过诱导Nrf2的表达、升高Bcl2的表达,降低Bax的表达起到了脑保护作用。

妇科再造丸含药血清对人子宫肌瘤细胞Bax和Bcl-2蛋白表达的影响

目的:探讨妇科再造丸(FKW)含药血清对体外培养人子宫肌瘤细胞凋亡相关蛋白Bax和B细胞淋巴瘤-2(Bcl-2)表达的影响。方法:对人子宫肌瘤细胞进行原代培养、传代并进行鉴定;60只SD雌性大鼠随机均分为空白组(无药血清组),FKW高、中、低剂量组和桂枝茯苓胶囊(GZFL)组,利用各组大鼠制备无药血清和含药血清分别作用于子宫肌瘤细胞72h时,采用WesternBlot检测细胞凋亡相关蛋白Bcl-2、Bax的表达。结果:培养的细胞经鉴定证实为人子宫肌瘤细胞;与无药血清组比较,FKW含药血清各剂量组均能增加Bax蛋白相对表达量,降低Bcl-2蛋白相对表达量(P<0.05或P<0.01),且作用随剂量的增加而有增强的趋势(P<0.05或P<0.01)。结论:FKW含药血清可影响人子宫肌瘤细胞凋亡相关蛋白Bcl-2、Bax的表达,这可能是FKW临床应用时缓解子宫肌瘤症状的机制之一。

大肠杆菌K5多糖对糖尿病性视网膜病变小鼠视网膜损伤的保护作用及组织中Bax、Nrf2水平的影响

目的探究并分析大肠杆菌K5多糖对糖尿病性视网膜病变(DR)小鼠视网膜损伤的保护作用及Bcl-2相关X蛋白(Bax)、核因子E2相关因子2(Nrf2)表达的影响。方法选取健康雄性C57BL/6小鼠120只,按照随机数字表法将其分为对照组和观察组,每组60只。两组小鼠进行糖尿病性视网膜病变造模处理,对照组小鼠又随机分为未处理组、处理(生理盐水)后1周组、处理(生理盐水)后2周组,每组20只。观察组小鼠又随机分为未处理组、处理(大肠杆菌K5多糖)后1周组、处理(大肠杆菌K5多糖)后2周组,每组20只。观察两组小鼠处理前、处理后1周、处理后2周视觉电生理指标及Bax、Nrf2蛋白表达水平。结果两组小鼠处理前波峰潜时、波峰振幅比较无显著差异(P>0.05),处理后,两组小鼠视觉电生理指标显著改善(P<0.05),观察组小鼠波峰潜时显著低于对照组(P<0.05),波峰振幅显著高于对照组(P<0.05);两组小鼠处理前视网膜节细胞、视网膜内网层细胞Bax蛋白水平比较无显著差异(P>0.05),处理后,两组小鼠视网膜节细胞、视网膜内网层细胞Bax蛋白水平显著降低,观察组视网膜节细胞、视网膜内网层细胞Bax蛋白水平明显低于对照组(P<0.05);两组小鼠处理前Nrf2蛋白水平比较无显著差异(P>0.05),处理后,两组小鼠Nrf2蛋白水平显著升高(P<0.05),观察组小鼠Nrf2蛋白水平明显高于对照组,差异具有统计学意义(P<0.05)。结论大肠杆菌K5多糖对DR小鼠视网膜损伤具有保护作用,同时能有效改善小鼠凋亡基因Bax及Nrf2的表达。

JTE-522-induced apoptosis in human gastric adenocarinoma cell line AGS cells by caspase activation accompanying cytochrome C release,membrane translocation of Bax and loss of mitochondrial membrane potential

AIM: To investigate the role of the mitochondrial pathway in JTE-522-induced apoptosis and to investigate the relationship between cytochrome C release, caspase activity and loss of mitochondrial membrane potential (Deltapsim). METHODS: Cell culture, cell counting, ELISA assay, TUNEL, flow cytometry, Western blot and fluorometric assay were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanism. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Caspases 8 and 9 were activated during apoptosis as judged by the appearance of cleavage products from procaspase and the caspase activities to cleave specific fluorogenic substrates. To elucidate whether the activation of caspases 8 and 9 was required for the apoptosis induction, we examined the effect of caspase-specific inhibitors on apoptosis. The results showed that caspase inhibitors significantly inhibited the apoptosis induced by JTE-522. In addition, the membrane translocation of Bax and cytosolic release of cytochrome C accompanying with the decrease of the uptake of Rhodamin 123, were detected at an early stage of apoptosis. Furthermore, Bax translocation, cytochrome C release, and caspase 9 activation were blocked by Z-VAD.fmk and Z-IETD-CHO. CONCLUSION: The present data indicate a crucial association between activation of caspases 8, 9, cytochrome C release, membrane translocation of Bax, loss of Deltapsim and JTE-522-induced apoptosis in AGS cells.

miR⁃21、ILK及Bax蛋白在CIN中的表达及临床意义

目的探究微小RNA⁃21(miR⁃21)、整合素连接激酶(ILK)、Bcl⁃2相关X蛋白(Bax)在宫颈上皮内瘤变(CIN)中的表达及临床意义。方法将2020年1月至2022年10月苏州科技城医院收诊并治疗的63例宫颈上皮内瘤变(CIN组)纳入研究,选择同期84名健康志愿者(对照组)共同参与本次研究,对比两组miR⁃21、ILK及bax蛋白的表达,分析宫颈组织中miR⁃21、ILK及Bax蛋白阳性表达与病理特征的关系。结果CIN组miR⁃21相对量高于对照组,差异有统计学意义(t=16.906,P<0.05);CIN组Bax蛋白阳性率低于对照组,差异有统计学意义(x^(2)=70.948,P<0.05);CIN组ILK阳性率高于对照组,差异有统计学意义(x^(2)=109.485,P<0.05);不同年龄阶段、不同组织类型的miR⁃21、Bax蛋白、ILK阳性表达率比较差异无统计学意义(P>0.05);而CIN分级越高,宫颈组织中ILK阳性率高,Bax阳性率越低,miR⁃21相对量越多,差异均有统计学意义(x^(2)=0.005,x^(2)=0.001,t=0.027,P<0.05)。结论miR⁃21、ILK、Bax蛋白均在CIN患者中异常表达,与病变细胞的凋亡密切关联。

Correlation of serum PDCD5 and Bax contents with the pathological features of tumor lesions in patients with lung cancer

Objective:To study the correlation of serum PDCD5 and Bax contents with the pathological features of tumor lesions in patients with lung cancer.Methods:Patients with lung cancer who underwent surgical treatment in West China Hospital between June 2014 and March 2017 were selected as the lung cancer group for the study, the serum specimens were collected before surgery, and the lung cancer lesion and adjacent lesion were taken after surgery;the healthy subjects who underwent physical examination in West China Hospital during the same period were selected as the control group, and the serum samples were taken during the physical examination;the contents of PDCD5 and Bax in serum as well as the contents of PDCD5 and Bax, and the expression of proliferation genes and invasion genes in the lung cancer lesion and adjacent lesion were measured.Results: PDCD5 and Bax contents in serum of lung cancer group were significantly lower than those of control group, PDCD5 and Bax contents in lung cancer lesion were significantly lower than those in adjacent lesion, and the PDCD5 and Bax contents in serum of patients with lung cancer were positively correlated with the PDCD5 and Bax contents in lung cancer lesion;C-myc, RACK1, CatL, MMP2 and N-cadherin mRNA expression in lung cancer lesion were significantly higher than those in adjacent lesion and negatively correlated with PDCD5 and Bax contents in serum whereas LAST1, PAQR3, TCF21, E-cadherin, TIMP1 and TIMP2 mRNA expression were significantly lower than those in adjacent lesion and positively correlated with PDCD5 and Bax contents in serum. Conclusion:The decrease of PDCD5 and Bax in serum of patients with lung cancer is closely related to the aggravation of cancer cell proliferation and invasion in tumor lesions.

急性脑梗死合并肺部感染病原菌耐药性及血清miR-5787、miR-133、Bax、Bcl-2表达水平

目的探讨急性脑梗死合并肺部感染(PI)患者病原菌耐药特点及血清微小核糖核酸-5787(miR-5787)、微小核糖核酸-133(miR-133)、B淋巴细胞瘤-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)表达水平。方法选取2021年1月-2023年8月峨眉山市人民医院收治的82例急性脑梗死合并PI患者作为A组,另选取同期收治的90例急性脑梗死无PI患者作为B组;分析A组患者病原菌分布特点及耐药性,比较两组血清miR-5787、miR-133、Bax、Bcl-2表达水平、红细胞免疫参数及炎性因子水平,并采用Pearson相关性分析血清miR-5787、miR-133、Bax、Bcl-2与红细胞免疫参数、炎性因子的相关性。结果82例急性脑梗死合并PI患者共分离出132株病原菌株,革兰阳性菌的占比为34.85%,主要为金黄色葡萄球菌,革兰阴性菌的占比为61.36%,主要为肺炎克雷伯菌,真菌的占比为3.79%;肺炎克雷伯菌对克林霉素、环丙沙星及磺胺甲噁唑/甲氧苄啶的耐药性较高,耐药率均为55.17%;金黄色葡萄球菌对阿莫西林、氨苄西林、青霉素、头孢他啶、环丙沙星的耐药性较高;与B组比较,A组血清miR-5787、miR-133、Bax水平均更高,血清Bcl-2水平更低(P<0.05);A组全血免疫黏附抑制因子(FEIR)及血清白细胞介素(IL)-17、肿瘤坏死因子-α(TNF-α)水平高于B组,全血协同肿瘤红细胞花环率(ATER)、自然肿瘤红细胞花环率(NTER)及血清IL-10水平低于B组(P<0.05);血清miR-5787、miR-133、Bax与全血FEIR、血清IL-17、TNF-α水平呈正相关,与全血ATER、NTER及血清IL-10水平呈负相关;血清Bcl-2与全血FEIR、血清IL-17、TNF-α水平呈负相关,与全血ATER、NTER及血清IL-10水平呈正相关(P<0.05)。结论急性脑梗死合并PI病原菌中占比较高的为肺炎克雷伯菌,且PI会导致患者机体促炎/抗炎机制失衡,抑制红细胞免疫功能,且血清miR-5787、miR-133、Bax、Bcl-2表达变化与患者红细胞免疫及炎症指标密切相关。

山慈菇通过PI3K/Akt信号通路影响乳腺癌MDA-MB-231细胞的增殖和凋亡

目的:研究山慈菇是否通过磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路影响乳腺癌MDA-MB-231细胞的增殖和凋亡。方法:将MDA-MB-231细胞分为对照组(C组)、山慈菇组(CAM组)、胰岛素生长因子1组(IGF-1组)、山慈菇+胰岛素生长因子1组(CAM+IGF-1组)。C组不加药物处理,CAM组加入10 mg/ml山慈菇处理,IGF-1组加入100μg/L IGF-1处理,CAM+IGF-1组加入10 mg/ml山慈菇和100μg/L IGF-1共处理。噻唑蓝(MTT)测定细胞增殖,流式细胞仪和Hoechst 33258染色测定细胞凋亡,Western blot法测定细胞活化的半胱天冬氨酸3(Cleaved-Caspase-3)、Bcl-2相关X蛋白(Bax)、B-细胞淋巴瘤因子(Bcl-2)、PI3K、磷酸化PI3K(p-PI3K)、Akt、p-Akt蛋白水平。结果:山慈菇抑制MDA-MB-231细胞增殖呈剂量和时间依赖性。与C组比较,CAM组MDA-MB-231细胞OD值降低(P<0.05),细胞凋亡率升高(P<0.05),Cleaved-Caspase-3、Bax蛋白水平升高(P<0.05),Bcl-2、p-PI3K、p-Akt蛋白水平降低(P<0.05);与CAM组比较,CAM+IGF-1组MDA-MB-231细胞OD值升高(P<0.05),细胞凋亡率降低(P<0.05),Cleaved-Caspase-3、Bax蛋白水平降低(P<0.05),Bcl-2、p-PI3K、p-Akt蛋白水平升高(P<0.05)。结论:山慈菇抑制乳腺癌细胞增殖、诱导乳腺癌细胞凋亡,其机制可能与山慈菇抑制PI3K/Akt信号通路活化有关。

生物钟基因Period2对卵巢癌裸鼠移植瘤生长抑制作用的机制研究

目的:探讨生物钟基因Period2对卵巢癌裸鼠移植瘤生长抑制的作用机制。方法:采用卵巢癌SKVO3细胞株构建裸鼠卵巢癌移植瘤,利用基因转染技术外源性导入重组基因质粒Period2。采用Real-time PCR法和Western bolt法检测移植瘤中Period2表达,治疗期间测量移植瘤体积。治疗后2周处死裸鼠,称取瘤重,采用Real-time PCR和Western blot法检测肿瘤组织中Bcl-2和Bax基因的表达。结果:外源性导入Period2基因在裸鼠移植瘤肿瘤组织中成功稳定表达。与对照组比较,Period2基因过表达组的肿瘤生长速度减慢,肿瘤体积和重量降低,抑瘤率升高,凋亡基因Bax表达明显升高,凋亡抑制基因Bcl-2表达明显降低,差异均有统计学意义(P<0.05)。结论:Period2可能通过下调肿瘤凋亡抑制基因Bcl-2,促进凋亡基因Bax表达,促进肿瘤细胞凋亡,从而抑制卵巢癌的生长转移,发挥抑瘤作用。

乙酰辅酶A合成酶2低表达人胃癌细胞株MGC80-3的顺铂敏感性观察

目的观察乙酰辅酶A合成酶2(ACSS2)低表达人胃癌细胞株MGC80-3的顺铂敏感性,并进一步探讨其可能作用机制。方法 采用Western blotting法检测人胃癌细胞株MGC80-3、AGS、SGC-7901、NCI-N87以及正常胃黏膜上皮细胞系RGM-1中ACSS2蛋白。对数生长期MGC80-3细胞分为A、B、C、D组,B及D组分别加入10 μL脂质体Lipofectamine 2000+5 μL siRNA-ACSS2转染, C组加入及A组分别加入50 μL无血清纯RPMI1640培养基+5 μL 阴性对照(NC);A组、B组均培养72 h,C组培养48 h时加入5 μg/mL的顺铂(DDP),D组转染48 h时加入5 μg/mL的DDP。培养24 h时,CCK8法检测各组细胞增殖活力,培养48、72 h时流式细胞分析术测算各组细胞早晚期凋亡率,Western blotting法检测各组细胞ACSS2、p65、p-p65、B淋巴细胞瘤-2蛋白(Bcl-2)、B细胞淋巴瘤-特大型蛋白(Bcl-xL)、凋亡蛋白Bcl-2相关的X蛋白(Bax)。结果 MGC80-3、AGS、SGC-7901、NCI-N87及RGM-1细胞ACSS2蛋白相对表达量分别为0.80± 0.03 、0.62±0.04、0.60±0.04、0.72±0.05、0.41±0.04。与RGM-1细胞比较,MGC80-3、AGS、SGC-7901、NCI-N87细胞ACSS2相对表达量升高( P 均<0.01)。A、B、C、D组细胞增殖活力分别为98.67%±4.89%、92.67%±7.86%、62.67%± 4.05%、34.67%±7.61%,A组与B组相比无明显差异( P >0.05);与A组比较,C组细胞增殖活力下降( P < 0.001 );与B、C组比较,D组细胞增殖活力均下降( P 均< 0.001 )。A、B、C、D组早期细胞凋亡率分别为1.06%±0.02%、1.13%±0.08%、7.62%±0.55%、14.33%±0.21%,晚期细胞凋亡率分别为8.01%± 0.05%、8.13%±0.06%、10.81%±0.43%、15.98±0.18%。与A组比较,C、D组细胞早、晚期凋亡率均升高( P 均<0.05);与B组比较,D组早、晚期凋亡率均升高( P 均<0.05);与C组比较,D组细胞早、晚期凋亡率均升高( P 均< 0.05 )。与A组比较,C组p-p65、Bax蛋白相对表达量升高,Bcl-2、Bcl-xL相对表达量降低( P 均<0.05);与B组比较,D组细胞p65、p-p65、Bcl-2、Bcl-xL相对表达量均降低、Bax相对表达量升高( P 均<0.05)。结论 ACSS2低表达的MGC80-3细胞DDP敏感性升高。ACSS2可能通过调控NF-κB信号通路及Bcl-2、Bcl-xL、Bax蛋白的表达提高MGC80-3细胞的DDP敏感性。

Effect of Lichong decoction on expression of Bcl-2 and Bcl-2-associated X protein mRNAs in hysteromyoma model rat

OBJECTIVE:To study on effects of Lichong decoction on expression of apoptosis-controlling genes,Bcl-2 and Bcl-2-associated X protein(Bax) mRNAs in hysteromyoma tissue of the hysteromyoma model rat.METHODS:Fifty Wistar female rats were randomly divided into a normal group,a model group,a Lichong decoction group,a Guizifuling capsule group and a Mifepristone group.The hysteromyoma rat model was established by intraperitoneal injection of exogenous estrin and progestogens.Pathological examination of uterine tissue,uterine coefficient and uterine transverse diameter were made under optic microscope and expressions of Bcl-2 and Bax mRNAs in uterine tissue in the groups were detected with real-time fluorescent quantitative polymerase chain reaction(PCR) technique.RESULTS:After treatment,under microscope it was found that in the Lichong decoction group myometrium thinned,muscle fiber slightly overgrowth or long and thin,regular arrangement,inserting phenomenon of inner circular muscle and external longitudinal muscle was occasionally or not seen in the Lichong decoction group.The uterine coefficient and the uterine transverse diameter significantly decreased(P<0.01),and Bcl-2 mRNA expression significantly decreased(P<0.01) and Bax mRNA expression significantly increased in hysteromyoma tissue(P<0.01) in the Lichong decoction group as compared with the model group.CONCLUSION:Therapeutic effects of Lichong decoction on hysteromyoma is related with decrease of Bcl-2 mRNA expression and increase of Bax mRNA expression.

Paclitaxel induces apoptosis in human gastric carcinoma cells

AIM;To investigate the apoptosis in gastric cancer cells induced by paclitaxel,and the relation between this apoptosis and expression of Bcl-2 and Bax. METHODS:In in vitro experiments,MTT assay was used to determine the cell growth inhibitory rate.Transmission electron microscope and TUNEL staining method were used to quantitatively and qualitively detect the apoptosis status of gastric cancer cell line SGC-7901 before and after the paclitaxel treatment.Immunohistochemical staining was used to detect the expression of apoptosis-regulated gene Bcl-2 and Bax. RESULTS:Paclitaxel inhibited the growth of gastric cancer cell line SGC-7901 in a dose-and time-dependent manner. Paclitaxel induced SGC-7901 cells to undergo apoptosis with typically apoptotic characteristics,including morphological changes of chromatin condensation,chromatin crescent formation,nucleus fragmentation and apoptotic body formation.Paclitaxel could reduce the expression of apoptosis-regulated gene Bcl-2,and improve the expression of apoptosis-regulated gene Bax. CONCLUSION:Paclitaxel is able to induce the apoptosis in gastric cancer.This apoptosis may be mediated by down- expression of apoptosis-regulated gene Bcl-2 and up- expression of apoptosis-regulated gene Bax.

Baicalin extracted from Huangqin(Radix Scutellariae Baicalensis) induces apoptosis in gastric cancer cells by regulating B cell lymphoma(Bcl-2)/Bcl-2-associated X protein and activating caspase-3 and caspase-9

OBJECTIVE:To evaluate the effects of baicalin in human gastric cancer cells, including apoptosis-inducing effects, and to investigate its underlying mechanisms of action.METHODS:Cell proliferation and apoptosis assays were performed to investigate the anti-proliferation effects of baicalin in human gastric cancer BGC-823 and MGC-803 cells.Real time-quantitative polymerase chain reaction and Western blotting analysis were performed to elucidate the molecular mechanisms underlying the anti-tumor properties of baicalin.RESULTS:In BGC-823 and MGC-803 gastric cancer cells treated with 80, 120, and 160 μmol/L baicalin for 48 h, a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay showed that baicalin significantly inhibited cell proliferation in a dose-dependent manner, while flow cytometric analysis demonstrated that baicalin could induce apoptosis, also in a dose-dependent manner.Moreover, baicalin up-regulated the expression of caspase-3, caspase-9, and B cell lymphoma(Bcl-2)-associated X protein and down-regulated the expression of Bcl-2 at both the m RNA and protein level.CONCLUSION:Baicalin has potential as a therapeutic agent for gastric cancer by inducing apoptosis in cancer cells.

Effect of quercetin on the expression of Bcl-2/Bax apoptotic proteins in endometrial cells of lipopolysaccharide-induced-abortion mice

OBJECTIVE: To explore the effect of quercetin on the expressions of Bcl-2/Bax apoptotic proteins in endometrial cells in mice with abortion induced by lipopolysaccharide.METHODS: For in vivo experiment, twenty five Kunming mice were randomly divided into five groups at day 4 of pregnancy, with 5 mice per group. The mice were treated with lipopolysaccharide(LPS)through tail vein intravenous injection at day 4 of pregnancy, followed by different concentrations of quercetin by oral gavage consecutively at days 5 to6 of pregnancy. On day 7 of gestation, the mice were sacrificed and the histopathological changes of the uterus tissues were observed. Immunohistochemical staining was applied to the detection of Bcl-2/Bax apoptotic proteins in the endometrial cells. For in vitro experiment, the primary endometrial cells werecultured using a uterus tissue mass culturing method sampled at day 4.5 of pregnancy. The cells were treated with LPS with or without different dosages of quercetin, respectively, for 12 h after 80% confluence. The expression of Bcl-2/Bax apoptotic proteins were detected by western blotting.RESULTS: Both the in vivo and in vitro experiments showed decreased expression of Bcl-2 and enhanced expression of Bax after LPS treatment, leading to a decreased Bcl-2/Bax ratio. The expression of Bcl-2 significantly increased while the expression of Bax was significantly elevated, in the LPS plus quercetin group compared to the LPS only group.CONCLUSION: These results suggest that quercetin has protective effect by partially regulating the expression of Bcl-2/Bax proteins, which in turn inhibits endometrial cell apoptosis and benefits the embryo implantation.

Effect of heparin on apoptosis in human nasopharyngeal carcinoma CNE2 cells

In order to study the mechanism of the effect of heparin on apoptosis in carcinoma cells, the nasopharyngeal carcinoma cell line CNE2 was used to identify the effect of heparin on apoptosis associated with the expression of c-myc, bax, bcl-2 proteins by use of Hoechst 33258 staining, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), agarose gel electrophoresis, and flow cytometry, as well as Western blot analysis. The results showed that heparin induced apoptosis of CNE2 cells including the morphologic changes such as reduction in the volume, and the nuclear chromatin condensation, as well as the 'ladder pattern' revealed by agarose gel electrophoresis of DNA in a concentration-dependent manner. The number of TUNEL-positive cells was dramatically increased to 33.6+/-1.2% from 2.8+/-0.3% by treatment with heparin in different concentrations (10 to approximately 40 kU/L). The apoptotic index was increased to 32.5% from 3.5% by detecting SubG1 peaks on flow cytometry. Western blot analysis showed that levels of bcl-2, bax and c-myc were significantly overexpressed by treatment with the increase of heparin concentrations. These results suggest that heparin induces apoptosis of CNE2 cells, which may be regulated by differential expression of apoptosis-related genes.

Effects of erythropoietin on the expression of tumor necrosis factor-alpha and Bax after facial nerve axotomy in rats

This study sought to evaluate the effect of high-dose erythropoietin （EPO; 5 000 IU/kg） on the expression of tumor necrosis factor-alpha （TNF-α） and Bax in the facial nucleus after facial nerve transection in rats. A total of 42 Wistar rats of both genders were used in this study, and 40 rats were randomly divided into 2 groups： EPO group and model group. The EPO group was treated with EPO once a day for 5 days at a dose of 5 000 IU/kg body weight. The model group was treated with saline of the same amount. At day 3 after EPO （or saline） treatment, the right facial nerves of the 40 rats were transected at the level of the stylomastoid foramen, with the left sides untreated. The remaining 2 rats that did not undergo axotomy served as the control group. The surviving motor neurons in operated rats were counted in coronal paraffin sections of the facial nucleus. The expression of TNF-a and Bax in the facial nucleus was detected by immunohistochemical staining at days 3, 7, 14, 21, and 28 after axotomy. At days 14, 21, and 28 after facial nerve axotomy, a significantly greater proportion of facial motor neurons survived in the EPO group than in the model group. After axotomy, the expression of TNF-a and Bax increased in motor neurons in both the EPO and the model groups. TNF-o expression reached its peak level at day 14 after axotomy, while Bax expression reached its peak level at day 21. TNF-α expression was much lower in the EPO group than in the model group at all time points. No significant difference in Bax expression was found between the EPO and the model groups. These results indicate that high-dose EPO treatment attenuates the increase in TNF-α expression in the facial nucleus and reduces the loss of motor neurons after facial nerve transection in rats. However, high-dose EPO treatment has little effect on Bax expression.

Yinchenhao decoction attenuates obstructive jaundice-induced liver injury and hepatocyte apoptosis by suppressing protein kinase RNA-like endoplasmic reticulum kinase-induced pathway

BACKGROUND Chronic biliary obstruction results in ischemia and hypoxia of hepatocytes,and leads to apoptosis.Apoptosis is very important in regulating the homeostasis of the hepatobiliary system.Endoplasmic reticulum(ER)stress is one of the signaling pathways that induce apoptosis.Moreover,the protein kinase RNA-like endoplasmic reticulum kinase(PERK)-induced apoptotic pathway is the main way;but its role in liver injury remains unclear.Yinchenhao decoction(YCHD)is a traditional Chinese medicine formula that alleviates liver injury and apoptosis,yet its mechanism is unknown.We undertook this study to investigate the effects of YCHD on the expression of ER stress proteins and hepatocyte apoptosis in rats with obstructive jaundice(OJ).AIM To investigate whether YCHD can attenuate OJ-induced liver injury and hepatocyte apoptosis by inhibiting the PERK-CCAAT/enhancer-binding protein homologous protein(CHOP)-growth arrest and DNA damage-inducible protein 34(GADD34)pathway and B cell lymphoma/leukemia-2 related X protein(Bax)/B cell lymphoma/leukemia-2(Bcl-2)ratio.METHODS For in vivo experiments,30 rats were divided into three groups:control group,OJ model group,and YCHD-treated group.Blood was collected to detect the indicators of liver function,and liver tissues were used for histological analysis.For in vitro experiments,30 rats were divided into three groups:G1,G2,and G3.The rats in group G1 had their bile duct exposed without ligation,the rats in group G2 underwent total bile duct ligation,and the rats in group G3 were given a gavage of YCHD.According to the serum pharmacology,serum was extracted and centrifuged from the rat blood to cultivate the BRL-3A cells.Terminal deoxynucleotidyl transferase mediated dUTP nick end-labelling(TUNEL)assay was used to detect BRL-3A hepatocyte apoptosis.Alanine aminotransferase(ALT)and aspartate transaminase(AST)levels in the medium were detected.Western blot and quantitative real-time polymerase chain reaction(qRT-PCR)analyses were used to detect protein and gene expression levels of PERK,CHOP,GADD34,Bax,and Bcl-2 in the liver tissues and BRL-3A cells.RESULTS Biochemical assays and haematoxylin and eosin staining suggested severe liver function injury and liver tissue structure damage in the OJ model group.The TUNEL assay showed that massive BRL-3A rat hepatocyte apoptosis was induced by OJ.Elevated ALT and AST levels in the medium also demonstrated that hepatocytes could be destroyed by OJ.Western blot or qRT-PCR analyses showed that the protein and mRNA expression levels of PERK,CHOP,and GADD34 were significantly increased both in the rat liver tissue and BRL-3A rat hepatocytes by OJ.The Bax and Bcl-2 levels were increased,and the Bax/Bcl-2 ratio was also increased.When YCHD was used,the PERK,CHOP,GADD34,and Bax levels quickly decreased,while the Bcl-2 levels increased,and the Bax/Bcl-2 ratio decreased.CONCLUSION OJ-induced liver injury and hepatocyte apoptosis are associated with the activation of the PERK-CHOP-GADD34 pathway and increased Bax/Bcl-2 ratio.YCHD can attenuate these changes.

Effect of Icariin on apoptosis and expression of Fas,Fas ligand,B cell lymphoma,and Bcl-2-associated X protein in CD4＋ T lymphocytes from patients with ankylosing spondylitis

OBJECTIVE:To investigate the effects of icariin on apoptosis and the expression of Fas, Fas ligand(Fas L), B cell lymphoma(Bcl-2), and Bcl-2-associated X protein(Bax) in CD4+ T lymphocytes from patients with ankylosing spondylitis.METHODS:Primary cultures of peripheral blood CD4+ T lymphocytes were established and treated with icariin at high, medium, and low doses(0.5,0.25, and 0.125 mg/mL).Sulfasalazine treated and helthy cells were used as controls.Apoptosis of treated cells was determined by flow cytometry.Reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assays were used to determine the effects of icariin on the expression of Fas, Fas L, Bcl-2, and Bax.The activity of caspase 8 and caspase 3 was determined by a colorimetric assay.RESULTS:The m RNA and protein expression of Fas,and activity of caspase 8 and caspase 3 in CD4+ T lymphocytes were increased by icariin(P < 0.05).Conversely, the m RNA and protein expression of Bcl-2 was decreased(P < 0.05).The expression of Fas L and Bax were not significantly different between groups.The proapoptotic effects of icariin were dose-dependent.CONCLUSION:Icariin induces the apoptosis of CD4 + T cells from patients with AS comparing to normal control.Therefore, the induction of apoptosis may be the likely mechanism of action of icariin's antirheumatics activities.

拉贝洛尔辅助硫酸镁对早发重度子痫前期患者胎盘组织中Fas、Bax表达及动脉血流指标影响观察

目的:观察拉贝洛尔辅助硫酸镁对早发重度子痫前期患者胎盘组织中凋亡相关因子(Fas)及其配体(Fas L)、B淋巴细胞瘤-2相关X蛋白(Bax)表达及动脉血流指标影响。方法:早发重度子痫前期患者144例随机分为对照组和观察组各72例。对照组予硫酸镁治疗,观察组在对照组基础上加用拉贝洛尔治疗。两组均治疗10 d,并随访至分娩。比较两组患者治疗前后血压、动脉血流指标以及血管内皮功能变化,观察两组妊娠结局及胎盘组织凋亡分子表达。结果:观察组胎盘组织中Fas、Fas L、Bax表达均明显低于对照组(P<0.05);且各类妊娠不良结局发生率均低于对照组(P<0.05)。治疗后,两组患者的收缩压、舒张压,搏动指数、阻力指数,以及血清高迁移率族蛋白B1、内皮素及血管性假血友病性因子水平均较治疗前显著降低,且观察组各项指标均明显低于对照组(P<0.05)。结论:拉贝洛尔辅助硫酸镁可显著降低早发重度子痫前期患者胎盘组织中Fas、Fas L、Baxs表达,降低血压,改善血管内皮功能及动脉血流指标,最终改善妊娠结局。

微小RNA-141-3p靶向调控含CUE结构域蛋白2基因对雨蛙素诱导的大鼠胰腺腺泡细胞损伤的影响

目的 探讨微小RNA-141-3p(miR-141-3p)靶向调控含CUE结构域蛋白2(CUEDC2)对雨蛙素诱导的大鼠胰腺腺泡细胞损伤的影响。方法 2018年7月至2019年12月,从美国ATCC购买大鼠胰腺腺泡细胞AR42J。100 nm/L雨蛙素(CAE)刺激大鼠胰腺腺泡细胞AR42J 6 h建立急性胰腺炎细胞模型。实时荧光定量PCR(qRT-PCR)和蛋白质印迹法(Western blotting)检测雨蛙素干预后AR42J细胞中miR-141-3p和CUEDC2的表达水平。双荧光素酶报告基因实验和蛋白质印迹法验证miR-141-3p和CUEDC2的靶向关系。利用脂质体转染法将miR-141-3p抑制物(anti-miR-141-3p)、miRNA抑制物阴性对照(anti-miR-NC)、CUEDC2过表达载体(pcDNA-CUEDC2)、空载体(pcDNA)分别转染AR42J细胞,经雨蛙素干预处理后,流式细胞术检测细胞凋亡,蛋白质印迹法检测B细胞淋巴瘤/白血病-2(Bcl-2)和Bcl-2相关X蛋白(Bax)的表达水平,酶联免疫吸附试验(Elisa)试剂盒检测细胞培养上清中肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平。结果 雨蛙素干预处理后AR42J细胞中miR-141-3p的表达水平显著升高[(2.43±0.24)比(1.00±0.09)],CUEDC2的表达水平显著降低。miR-141-3p靶向负性调控CUEDC2表达。与转anti-miR-NC和雨蛙素协同处理组比较,转染anti-miR-141-3p和雨蛙素协同处理组AR42J细胞凋亡率显著增加[(27.48±2.33)%比(15.64±1.51)%],Bax蛋白的表达显著增加,Bcl-2蛋白的表达显著降低,TNF-α[(136.54±14.58)ng/L比(226.48±20.54)ng/L]和IL-6[(115.89±11.65)ng/L比(193.47±18.63)ng/L]的分泌显著降低;与转染pcDNA和雨蛙素协同处理组比较,转染pcDNA-CUEDC2和雨蛙素协同处理组AR42J细胞凋亡率显著增加[(23.87±2.35)%比(14.23±1.44)%],Bax蛋白的表达显著增加,Bcl-2蛋白的表达显著降低,TNF-α[(158.74±15.32)ng/L比(236.87±18.66)ng/L]和IL-6[(135.77±14.97)ng/L比(189.67±17.32)ng/L]的分泌显著降低;转染si-CUEDC2可逆转转染anti-miR-141-3p对雨蛙素诱导的大鼠胰腺腺泡细胞损伤的影响。结论 抑制miR-141-3p通过靶向CUEDC2可促进急性胰腺炎腺泡细胞的凋亡,抑制TNF-α和IL-6分泌,从而减轻雨蛙素诱导的大鼠胰腺腺泡细胞损伤。