The Protective Effects of Flavonoids from Scutellaria Baicalensis Georgi Stems and Leaves on Oligodendrocyte Damage Induced by Aβ1-42

Aim: This study aimed to investigate the protective effects of flavonoids from the stem and leaves of Scutellaria baicalensis Georgi (SSFs) against Aβ1-42-induced oligodendrocytes (OL) damage. Methods: Immunofluorescence was used for the detection of myelin-associated glycoprotein (MAG), a characteristic protein of rat oligodendrocytes (OLN-93 cells). To evaluate the potential protective effects of SSFs on OLN-93 cells injured by Aβ1-42, an injury model was established by subjecting OLN-93 cells to Aβ1-42 exposed. Cell morphology was examined using an inverted microscope, while cell viability was assessed using the colorimetric method of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Additionally, lactate dehydrogenase (LDH) was measured using the pyruvic acid reduction assay. The Ginkgo biloba leaf extract (GBE) injection was used as a positive control. Results: A total of >95% of the MAG immunofluorescence-positive cells were identified as oligodendrocytes. Gradually increasing concentrations of SSFs impaired the cells, and the maximum nondetrimental dose for OLN-93 cells was 75 mg/L. This study assessed the effects of SSFs on OLN-93 cells damaged by Aβ1-42. The results indicated that SSFs significantly improved OLN-93 cell morphological abnormal changes, increased the OLN-93 cell survival rate, and reduced LDH release. Conclusion: SSFs can alleviate Aβ1-42-induced damage of OL.

Pretreatment with scutellaria baicalensis stem-leaf total flavonoid prevents cerebral ischemia-reperfusion injury

Pretreatment with scutellaria baicalensis stem-leaf total flavonoid has protective effects against ischemia and attenuates myocardial ischemia-reperfusion injury. In this study, rats were given scutellaria baicalensis stem-leaf total flavonoid intragastrically at 50, 100, and 200 mg/kg per day for 7 days before focal cerebral ischemia-reperfusion injury models were established using the suture method. We then determined the protective effects of scutellaria baicalensis stem-leaf total flavon- oid pretreatment on focal cerebral ischemia-reperfusion injury. Results showed that neurological deficit scores increased, infarct volumes enlarged, apoptosis increased and Bcl-2 and Bax protein expression were upregulated at 24 hours after reperfusion. Pretreatment with scutellaria baicalensis stem-leaf total flavonoid at any dose lowered the neurological deficit scores, reduced the infarct volume, prevented apoptosis in hippocampal cells, attenuated neuronal and blood-brain barrier damage and upregulated Bcl-2 protein expression but inhibited Bax protein expression. Doses of 100 and 200 mg/kg were the most efficacious. Our findings indicate that pretreatment with scutel- laria baicalensis stem-leaf total flavonoid at 100 and 200 mg/kg can improve the neurological func- tions and have preventive and protective roles after focal cerebral ischemia-reperfusion injury.

Pretreatment with scutellaria baicalensis stem-leaf total flavonoid protects against cerebral ischemia/reperfusion injury in hippocampal neurons

Previous experimental studies have shown that cerebral infarction can be effectively reduced following treatment with scutellaria baicalensis stem-leaf total flavonoid （SSTF）. However, the mechanism of action of SSTF as a preventive drug to treat cerebral infarction remains unclear. In this study, Sprague-Dawley rats were pretreated with 50, 100, 200 mg/kg SSTF via intragastric ad- ministration for 1 week prior to the establishment of focal cerebral ischemia/reperfusion injury. The results showed that pretreatment with SSTF effectively improved neurological function, reduced brain water content and the permeability of blood vessels, ameliorated ischemia-induced morphology changes in hippocampal microvessels, down-regulated Fas and FasL protein expression, elevated the activity of superoxide dismutase and glutathione peroxidase, and decreased malondialdehyde content. In contrast to low-dose SSTF pretreatment, the above changes were most obvious after pretreatment with moderateand high-doses of SSTF. Experimental findings indicate that SSTF pretreatment can exert protective effects on the brain against cerebral ischemia/reperfusion injury. The underlying mechanisms may involve reducing brain water content, increasing microvascular recanalization, inhibiting the apoptosis of hippocampal neurons, and attenuating free radical damage.

Pharmacological Activity and Application of Roots, Stems and Leaves of Scutellaria baicalensis Georgi

The root of Scutellaria baicalensis Georgi is traditionally used as medicine,and it has been confirmed that S.baicalensis Georgi has flavonoid chemical constituents,pharmacological activity and cosmetic efficacy.With the extensive application of S.baicalensis Georgi roots,the resource of S.baicalensis Georgi has been increasingly short.The above-ground part of stems and leaves of S.baicalensis Georgi has also been gradually recognized and developed.Studies have found that the chemical constituents from stems and leaves of S.baicalensis Georgi are also a group of flavonoids with a lot of pharmacological activity and have a great application value.Based on this,the present review will be reported on the chemical constituents and application of the roots,stems and leaves of S.baicalensis Georgi.

Effects of compatibility of Scutellaria baicalensis stems and Polygonum cuspidatum on TRPV1 expression and inflammatory cytokines in rats with acute lung injury

Objective:The protective effect of Scutellaria baicalensis Stems and Polygonum Cuspidatum compatibility on lipopolysaccharide(LPS)-induced acute lung injury(ALI)in rats was studied by observing the expression of TRPV1 and inflammatory cytokines.Methods:48 male SD rats were randomly divided into 6 groups:control group,model group,dexamethasone group(5mg/kg)and Scutellaria baicalensis Stems-Polygonum Cuspidatum(3.5,7 and 14g/kg).The administration group was gavaged for 7 days,and the control group and model group were given the same amount of 0.9%sodium chloride.On the 8th day,except the control group,rats in other groups were injected with 8mg/kg LPS through caudal vein to induce Ali model.Take the rat lung tissue 6 hours after modeling,and calculate the wet/dry weight ratio(W/D)of the rat lung tissue;HE staining to observe the pathological changes of lung tissue;Determine the content of tumor necrosis factor-α(TNF-α)and interleukin-1β(1L-1β)in alveolar lavage fluid(BALF)and the activity of superoxide dismutase(SOD)in serum;Detect the mRNA and protein expression levels of TRPV1 receptor in rat lung tissue.Results:Compared with the model group,Scutellaria baicalensis Stems-Polygonum Cuspidatum can significantly reduce the damage of lung tissue structure and bleeding state,W/D value and TNF-α、IL-1βThe content of TRPV1 decreased,the level of SOD increased,and the mRNA and protein expression of TRPV1 receptor decreased.Conclusion:The combination of Scutellaria baicalensis Stems-Polygonum has a protective effect on acute lung injury in rats,and its mechanism may be related to down-regulating the expression of TRPV1 and inhibiting the levels of TNF-αand IL-1βin inflammatory cells.

Inhibition of Flavonoids from Scutellaria baicalensis Stems and Leaves against Abnormal Formation of PHF and Regulation of PP in OA Induced Brain Injury in Rats

[Objectives] To explore the inhibitory effects of flavonoids from Scutellaria baicalensis Georgi stems and leaves against abnormal formation of PHF and regulation mechanism of protein phosphatase(PP) in okadaic acid(OA) induced brain injury in rats.[Methods]Male Sprague-Dawley rats were treated with OA(200 ng/kg) in the lateral ventricle to establish a memory impairment model,and Morris water maze was used to screen the memory impairment model.Rats were intragastrically administered with 25,50 and 100 mg/kg SSF daily for 36 d.The protein expressions of PHF,PP1,PP2A-Cα,PP2A-Cβ,PP2 CA and PP2 CB in rat cortex and hippocampus were determined by Western blot.Ginkgo biloba leaf flavonoids were used as positive control drugs.[Results]Compared with the sham-operated rats,the expression of PHF protein in the cortex and hippocampus of the model group was significantly increased(P < 0.01);protein expression of PP2A-Cα and PP2A-Cβin the cortex and hippocampus,and PP2 CB in the hippocampus were significantly decreased(P < 0.05);protein expression of PP2 CA and PP2 CB in the cortex was significantly increased(P < 0.01);protein expression of PP1 in the cortex was significantly decreased(P < 0.01).Compared with the model rats,25,50 and 100 mg/kg SSF 36 d reversed the protein expression of PHF,PP2A-Cα and PP2ACβ in rat cortex and hippocampus and expression of PP1 in rat cortex induced by OA,but had no significant effect on the protein expression of PP2 CA and PP2 CB,G.biloba flavonoids also showed similar results to SSF.[Conclusions] SSF can significantly inhibit the abnormal formation of PHF in rat brain induced by OA.The inhibitory effect may be related to the regulation of PP1,PP2A-Cα and PP2A-Cβ protein expression in cerebral cortex and hippocampus by SSF,but less related to the regulation of PP2 CA and PP2 CB.

黄芩茎叶葡萄糖醛酸水解酶提取工艺、酶学性质、实际应用研究

目的研究黄芩茎叶葡萄糖醛酸水解酶(sbslGUS)提取工艺、酶学性质、实际应用。方法在单因素试验基础上,以粒度、加水量、提取时间、提取次数为影响因素,酶活性为评价指标,正交试验优化提取工艺。分析底物(黄芩苷)浓度与酶解速度的关系,计算V_(max)、K_(m),考察pH值、温度、金属离子对酶活性的影响,评价pH稳定性、热稳定性。sbslGUS酶解黄芩苷以制备黄芩素,考察pH值、温度、反应时间、底物初始浓度、加酶量对转化率的影响。结果最优提取工艺为粒度40目,加水量10倍,提取时间15 min,提取次数3次。酶解符合酶促反应动力学,K_(m)为0.0063 mol/L,V_(max)为70.42μmol/h,在pH值6.0、温度45℃、金属离子100 mmol/L Cu2+下酶活性最强,sbslGUS在pH值4.0~7.0、温度4~30℃范围内稳定性良好。最优制备工艺为pH值6.0,温度45℃,反应时间12 h以上,底物初始浓度67.2 mmol/L,加酶量1 mL/0.269 mmol黄芩苷,转化率为97.83%。结论sbslGUS酶解效率高,条件温和,可为获取黄芩素提供简便的制备方法,并且拓展了黄芩茎叶应用途径。

酶法制备野黄芩素的工艺研究

目的应用黄芩茎叶葡萄糖醛酸水解酶(Scutellaria baicalensis stems and leaves glucuronic hydrolase,sbsl GUS)酶解灯盏花中野黄芩苷制备野黄芩素,经分离纯化获得高纯度的野黄芩素提取物。方法采用正交试验优选灯盏花提取工艺参数;以野黄芩苷酶解转化率为指标,对酶用量、酶解pH值、温度、时间、抗氧化剂等进行考察,优选野黄芩素制备工艺参数;采用乙醇提取、活性炭脱色和分步结晶精制纯化粗提物。结果灯盏花提取工艺为灯盏花切段,加10倍水煎煮提取2次,每次1 h。野黄芩素制备工艺为sbsl GUS提取液和灯盏花水煎液加入量(以生药计)比为1∶10,加0.5%偏重亚硫酸钠,酶解pH值约6.0,温度约45℃,时间20 h,得到含量大于60%的野黄芩素粗提物。经分步结晶对粗提物进行纯化,用80%乙醇回流提取,活性炭脱色,浓缩、析晶,得到含量大于85%的野黄芩素提取物。结论该工艺应用sbsl GUS酶解技术制备野黄芩素,生产简便可行,为野黄芩素的生产制备提供了新的途径。

黄芩茎叶发酵黄酮的体外抑菌及抗炎作用研究

试验旨在探究黄芩茎叶发酵黄酮的抗菌抗炎效果,以大肠杆菌、沙门氏菌、志贺氏菌和金黄色葡萄球菌为试验菌种,测定最小抑菌浓度(MIC)、最小杀菌浓度(MBC)及抑菌圈大小。50只SPF级昆明小鼠随机分为5组,每组10只,除对照组(Con组)各组小鼠均腹腔注射脂多糖(LPS,4 mg/kg),Con组灌胃相同剂量的生理盐水,阳性药物组(LPS+DM组)灌喂0.5 mg/kg地塞米松,原药组(LPS+SBSL组)和发酵药物组(LPS+FSBSL组)灌喂8 mg/kg原药和发酵黄酮。试验期7 d。结果显示,发酵黄酮MIC、MBC值最低的分别是大肠杆菌和金黄色葡萄球菌;在250 g/L条件下,发酵黄酮对大肠杆菌、痢疾志贺氏菌和金黄色葡萄球菌具有中等敏感性。与LPS组相比,LPS+FSBSL组小鼠死亡率明显降低;LPS+SBSL组小鼠肝脏指数极显著高于LPS+FSBSL组(P<0.01),LPS组脾脏指数极显著高于Con组(P<0.01)。与LPS组比,LPS+DM组和LPS+SBSL组小鼠血清白细胞介素-6(IL-6)含量极显著升高(P<0.01);与LPS+DM组比,LPS+SBSL组和LPS+FSBSL组小鼠血清IL-6含量极显著升高(P<0.01)。与Con组比,LPS组小鼠血清肿瘤坏死因子-α(TNF-α)含量显著升高(P<0.05)。与LPS组相比,LPS+FSBSL组小鼠肺脏组织中IL-6含量显著升高(P<0.05)。与LPS+SBSL组相比,LPS+FSBSL组小鼠肺脏组织中白细胞介素-1β(IL-1β)含量显著升高(P<0.05)。研究表明,黄芩茎叶发酵黄酮对痢疾志贺氏菌和金黄色葡萄球菌均有较好的抑菌效果,安全剂量为8 mg/kg,对LPS感染后小鼠的死亡率具有抑制作用,同时可以调节LPS感染后小鼠血清和肺脏中炎症因子IL-6、IL-1β及TNF-α含量。

微波辅助热水浸提黄芩茎叶多糖的工艺优化

为确定黄芩茎叶多糖的最佳提取工艺,采用单因素逐级优化试验与响应面分析法,以提取液中的多糖含量为指标,优化微波辅助热水浸提黄芩茎叶多糖的提取工艺。试验结果表明:单因素逐级优化试验筛选获得黄芩茎叶多糖的最佳提取条件为:微波预处理时间5 min,微波预处理功率800 W,料液比1∶35(g/mL),水提时间4 h,水提温度100℃,在此条件下黄芩茎叶多糖的提取率为17.39%。通过响应面分析法优化后所得黄芩茎叶多糖的最佳提取条件为:微波预处理时间312 s,微波预处理功率800 W,料液比1∶34(g/mL),水提时间280 min,水提温度100℃,在此条件下黄芩茎叶多糖的提取率为17.49%。优化得到的黄芩茎叶多糖提取工艺操作简单,合理可行。

响应面法优化黄芩茎叶黄酮类成分提取工艺及其含量测定

采用单因素和响应面试验考察乙醇浓度、料液比、提取时间和提取温度对黄芩茎叶中黄酮成分提取量的影响;在此基础上,采用HPLC法测定了17个黄芩茎叶中黄酮成分的含量。结果表明,黄芩茎叶中黄酮成分的最佳提取工艺为乙醇浓度55%、料液比1∶50、提取时间60 min、提取温度50℃,应用最佳提取工艺黄酮成分提取量为24.43 mg/g;不同产地黄芩茎叶中黄酮成分的含量存在差异,含量范围为15.839~26.044 mg/g。响应面法优化黄芩茎叶中黄酮成分的提取工艺稳定、准确可靠,所建立的黄酮成分含量测定方法稳定可靠,可为黄芩茎叶的质量评价及深度开发研究提供参考。

SSTF上调miR-495表达减缓高糖诱导的视网膜神经节细胞氧化损伤和凋亡

目的探讨黄岑茎叶总黄酮(SSTF)对高糖诱导的视网膜神经节细胞(RGC-5)损伤的保护机制。方法根据处理方法不同将RGC-5细胞分成高糖(HG)组、对照(Con)组、HG+SSTF-低剂量(L)组、HG+SSTF-中剂量(M)组、HG+SSTF-高剂量(H)组、HG+miR-NC组、HG+miR-495组、HG+SSTF+anti-miR-NC组、HG+SSTF+anti-miR-495组,每组设置9复孔。检测MDA含量以及CAT、SOD活性。流式细胞术检测细胞凋亡。实时荧光定量PCR(qRT-PCR)检测miR-495表达。结果HG组RGC-5细胞凋亡率、MDA含量高于Con组(P<0.05),CAT、SOD的活性、miR-495表达低于Con组(P<0.05)。HG+SSTF-L组、HG+SSTF-M组、HG+SSTF-H组RGC-5细胞凋亡率、MDA含量低于HG组(P<0.05),CAT和SOD的活性、miR-495表达高于HG组(P<0.05)。HG+miR-495组RGC-5细胞凋亡率[(14.96±1.26)%]、MDA含量[(4.62±0.44)μmol/g]分别低于HG+miR-NC组[(33.28±2.25)%、(9.73±0.81)μmol/g],差异有统计学意义(P<0.05),CAT、SOD的活性、miR-495表达水平高于HG+miR-NC组(P<0.05)。HG+SSTF+anti-miR-495组RGC-5细胞凋亡率[(22.21±2.18%)]、MDA含量[(8.07±0.75)μmol/g]高于HG+SSTF+anti-miR-NC组[(9.19±0.84)%、(3.59±0.32)μmol/g],差异有统计学意义(P<0.05),CAT和SOD的活性、miR-495表达水平低于HG+SSTF+anti-miR-NC组(P<0.05)。结论SSTF通过上调miR-495表达减缓高糖诱导的视网膜神经节细胞氧化损伤和凋亡。

黄芩茎叶总黄酮对小鼠肉瘤S_(180)瘤株体内增殖的抑制作用

目的:初步分析黄芩茎叶总黄酮的抗肿瘤药效。方法:采用小鼠肉瘤S180模型,观察黄芩茎叶总黄酮腹腔注射给药对移植瘤重量的影响。结果:2次重复实验,3个剂量的黄芩茎叶总黄酮皆可抑制移植肉瘤S180瘤株的体内增殖,量效关系较显著,其中,100～200mg/kg黄芩茎叶总黄酮的抑瘤率>30%,与对照组比较差异有高度显著性和显著性(P<0.01,P<0.05),但其活性低于50mg/kg环磷酰胺(平均抑瘤率达90.50%)。结论:黄芩茎叶总黄酮具有高效低毒的抗肿瘤活性,有可能开发成为新的抗肿瘤药物。

黄芩茎叶总黄酮治疗2型糖尿病性高脂血症大鼠的实验研究

目的观察黄芩茎叶总黄酮(SSTF)对2型糖尿病性高脂血症大鼠血糖、血脂代谢及胰岛素抵抗的影响,探究其可能的作用机制。方法高脂饮食加小剂量链脲佐菌素(STZ)建立2型糖尿病合并高脂血症大鼠模型,观察SSTF对大鼠血糖、血脂、胰岛素敏感性指数、抗氧化能力等方面的影响。结果SSTF大、小剂量组(150,75mg·kg-1)能显著降低实验大鼠空腹血糖(FBG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)水平,不同程度的降低甘油三酯(TG)水平,显著升高高密度脂蛋白胆固醇(HDL-C)水平和胰岛素敏感性指数(ISI)。SSTF能显著降低血清和肝脏中丙二醛(MDA)的含量,显著增加血清超氧化物歧化酶(SOD)的活性,不同程度地提高血清谷胱甘肽(GSH)含量。结论黄芩茎叶总黄酮降糖、降脂作用确切,并能改善胰岛素抵抗作用。SSTF的作用机制可能和其显著的抗氧化作用有关。

黄芩茎叶总黄酮预处理对缺血再灌注心肌脂质过氧化损伤的保护作用

目的：探讨黄芩茎叶总黄酮(SSTF)预处理对缺血再灌注心肌脂质过氧化损伤的保护作用。方法：雄性大鼠分为SSTF预处理组、缺血再灌组和假手术组,缺血再灌术前给SSTF组大鼠SSTF(50、100、200mg·kg^1·d^(-1))灌胃,制备缺血再灌模型,分别测定心肌组织超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,血清乳酸脱氢酶(LDH)、肌酸激酶(CK)水平。结果：SSTF预处理可不同程度显著降低缺血再灌注引起的MDA含量,提高SOD活性,降低LDH、CK水平。结论：SSTF预处理能通过保护心肌抗氧化酶的活性,抵制脂质过氧化反应,减轻自由基对心肌的损害,对缺血再灌注心肌产生保护作用。

不同产地黄芩茎叶UPLC指纹图谱与化学模式识别研究

目的采用超高效液相色谱法(UPLC)建立不同产地黄芩茎叶指纹图谱,超高效液相色谱串联四极杆飞行时间质谱法(UPLC-QTOF/MS)快速鉴定共有峰,并结合化学模式识别研究,为黄芩地上茎叶资源循环利用与质量控制提供参考。方法采用UPLC,色谱条件为ACQUITY UPLC BEH C18(100mm×2.1mm,1.7μm)色谱柱,以乙腈-0.1%甲酸水为流动相梯度洗脱,流速0.4mL/min,检测波长254nm,柱温35℃。质谱检测采用负离子模式,电压3.0kV,离子源温度120℃,雾化温度400℃,雾化气800L/h。通过相似度评价结合聚类分析和主成分分析进行分析评价,并通过UPLC-QTOF/MS对黄芩茎叶共有峰进行鉴定。结果确定了不同产地黄芩茎叶药材的19个共有峰,建立了UPLC指纹图谱。10个产地11批黄芩茎叶样品的指纹图谱相似度均>0.9,各产地黄芩茎叶药材的相似度较高,聚类分析、主成分分析结果与相似度分析结果一致。结论本方法快速、准确、可靠、重复性好,黄芩茎叶UPLC指纹图谱的构建和化学模式的识别可更好的评价其质量,为黄芩地上部分资源价值的利用与质量的控制提供参考。

黄芩茎叶总黄酮对培养大鼠心肌细胞H_2O_2损伤的保护作用

目的:探讨黄芩茎叶总黄酮保护培养大鼠心肌细胞抗过氧化氢(H2O2)损伤。方法:应用H2O2培养大鼠心肌细胞损伤模型,采用乳酸脱氢酶(LDH)活性及MTT代谢率的测定等方法。结果:与H2O2损伤组比较,黄芩茎叶总黄酮保护组的LDH释放量显著减少(P<0.01),而MTT代谢率显著增高(P<0.01)。结论:黄芩茎叶总黄酮能保护培养大鼠心肌细胞,减轻自由基损伤。

黄芩茎叶总黄酮调节EAE大鼠Th17/Treg平衡的研究

目的观察黄芩茎叶总黄酮(Scutellaria baicalensisstem-leaf total flavonoids,SSTF)对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)大鼠疾病严重程度和Th17、Treg细胞的影响,初步探讨黄芩茎叶总黄酮对多发性硬化症(multiple sclerosis,MS)的治疗作用。方法将50只大鼠随机分为正常组、模型组、黄芩茎叶总黄酮高剂量组(200 mg/kg)、黄芩茎叶总黄酮低剂量组(100 mg/kg)和醋酸泼尼松组(6 mg/kg),建模后分别行神经功能评分。灌胃给药16 d后,大鼠脊髓组织行HE染色,ELISA法测定大鼠血清中IL-17、TGF-β水平,流式细胞术检测大鼠外周血Th17、Treg细胞数量。结果与模型组大鼠比较,治疗组大鼠平均神经功能评分降低(P<0.05),炎性细胞浸润减少(P<0.05);血清中IL-17水平显著降低(P<0.05),而TGF-β水平显著升高(P<0.05);外周血中Th17细胞比例明显降低(P<0.05),而Treg细胞比例显著升高(P<0.05)。结论黄芩茎叶总黄酮灌胃给药可以调节CD4+T淋巴细胞亚群Th17/Treg细胞平衡,对EAE大鼠有一定的保护作用。