Ramulus Cinnamomi extract attenuates neuroinflammatory responses via downregulating TLR4/MyD88 signaling pathway in BV2 cells

Ramulus Cinnamomi （RC）, a traditional Chinese herb, has been used to attenuate inflammatory responses. The purpose of this study was to investigate the effect of RC extract on lipopolysaccharide （LPS）-induced neuroinflammation in BV2 microglial cells and the underlying mechanisms involved. BV2 cells were incubated with normal medium （control group）, LPS, LPS plus 30 pg/mL RC extract, or LPS plus 100 pg/mL RC extract. The BV2 cell morphology was observed under an optical microscope and cell viability was detected by MTT assay. Nitric oxide level in BV2 cells was detected using Griess regents, and the levels of interleukin-6, interleukin-1 β, and tumor necrosis factor u in BV2 cells were determined by ELISA. The expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 proteins were detected by western blot assay. Compared with the LPS group, both 30 and 100 μg/mL RC extract had no significant effect on the viability of BV2 cells. The levels of nitric oxide, interleukin-6, interleukin-1β and tumor necrosis factor ct in BV2 cells were all significantly increased after LPS induction, and the levels were significantly reversed after treatment with 30 and 100 μg/mL RC extract. Furthermore, RC extract significantly inhibited the protein expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 in LPS-induced BV2 cells. Our findings suggest that RC extract alleviates neuroinflammation by downregulating the TLR4/MyD88 signaling pathway.

The extract of Ramulus Cinnamom attenuates inflammatory injury induced by LPS via regulation of TLR4/MyD88 signaling pathway in BV2 cells

OBJECTIVE To investigate the effects of ex-tract of Ramulus Cinnamom(RC)against LPS-induced inflammation in microglia.METHODS Activated microglia releases various pro-inflammatory cytokines to induce neuroinflammation in stroke.Lipopolysaccaride(LPS)is an endotoxin from the outer membrane of Gram-negative bacteria that activates microglia.MTT assay was used to observe the cell viability.The content of NO in supernatant was measured by Griess reagent.The levels of IL-1β,IL-6 and TNF-αin supernatant were detected by ELISA kits.The intracellular COX-2,TLR4,and My D88expression was assayed by Western blotting.RESULTS RC extract 30 and 100μg·m L-1significantly decreased the production of related inflammatory factors such as NO(P<0.05,P<0.01),IL-1β(P<0.01,P<0.01),IL-6(P<0.05,P<0.01)and TNF-α(P<0.01,P<0.01).Furthermore,RC extract significantly inhibited the COX-2,TLR4,and My D88 expression induced by LPS in BV2cells.CONCLUSION RC extract may have therapeutic potential for the improvement of neuroinflammation,and the mechanism may be involved in down-regulation of TLR4/My D88 inflammation pathway.

Advances in Toxicity of Ramulus et Uncus Uncariae

Ramulus et Uncus Uncariae is one of the 31 regional characteristic Chinese medicinal materials identified by Guangxi Zhuang Autonomous Region in 2021,with antihypertensive,antiarrhythmic,anti-cerebral ischemia,sedative,antithrombotic,anti-tumor and other pharmacological effects,and is neurotoxic and hepatotoxic.In this article,the chemical constituents,pharmacological effects,toxic effects,toxic manifestations in whole animals and mechanism of toxicity of Ramulus et Uncus Uncariae are reviewed.

Mechanisms of Cinnamomi Ramulus for coronary heart disease treatment:prediction based on network pharmacology

Chinese medicine.However,little is known about the potential mechanism.Elucidating the effective components and mechanism based on network pharmacology was our purpose.Methods:Traditional Chinese Medicine Systems Pharmacology was screened to collect the possible active ingredients and their CAS and SMILES was searched in Pubchem,and the related potential targets were further predicted in Swiss Target Prediction database.Coronary heart disease molecular members were gained from GeenCards database,and the predicted targets of Cinnamomi Ramulus for coronary heart disease’s treatment were selected by Wayne diagram.As for mechanism analysis,String was used to construct the protein-protein interactions,and DAVID was used to conduct the GO and KEGG analysis.Results:Through GO and KEGG analysis,we found that cAMP signaling pathway,estrogen signaling pathway,calcium signaling pathway was related with coronary heart disease.Using network-based systems biology,we predicted that 10 active ingredients in Cinnamomi Ramulus have the treating effects with 78 potential targets.PIK3CG,MAPK8,BCL-2,BAX,PRKACA,CASP3,CALM1,CALM2,CALM3,NOS2,NOS3 were mainly involved in the treating effects of Cinnamomi Ramulus.Conclusion:Cinnamomi Ramulus may treat coronary heart disease through cAMP signaling pathway,estrogen signaling pathway,calcium signaling pathway.PIK3CG,MAPK8,BCL-2,BAX,PRKACA,CASP3,CALM1,CALM2,CALM3,NOS2,NOS3 were supposed to be considerable targets for treating coronary heart disease.

Research progress on the effect of the combination of Jing acupoints bloodletting therapy and Sangzhi (Mori Ramulus) on shoulder-hand syndrome after stroke

Shoulder-hand syndrome(SHS)is one of the common complications of ischemic stroke.The pathogenesis is not completely clear and the therapeutic effects are not very satisfactory.As one of the Five-Shu acupoints(the general terms of acupoints that the twelve meridians are located below the elbow and knee of the body),Jing acupoints is distributed at the end of fingers and toes where the twelve meridians of the human body pass by,and has the functions of stimulating the meridians and dredging the channels and collaterals.For the effects of discharging neurons,promoting cerebral blood flow and improving the brain micro-circulation,Jing acupoints bloodletting therapy can effectively relieve the clinical symptoms of the patients with SHS after stroke.Sangzhi(Mori Ramulus),with the ability of dredging the meridian and relieving the pain,is also has certain treatment functions to the SHS.In clinical practice,the combination of Jing acupoints bloodletting and Sangzhi(Mori Ramulus)have been widely used in the treatment of various diseases,and in terms of their mechanism of action,the combined treatment has a positive effect on post-stroke SHS,but there are few reports on this.Therefore,it is worth affirming the efficacy of combined treatment of SHS after stroke.This article elaborates the theoretical basis of Jing acupoints bloodletting on SHS after stroke,and the research progress of Sangzhi(Mori Ramulus)in treating SHS after stroke,which provide the theoretical guidance for the combination.

Quantification of seven phenylpropanoid compounds in Chinese Cinnamomi Cortex and Ramulus by HPLC

In the present study, we developed and validated a high-performance liquid chromatography method for the simultaneous determination of seven phenylpropanoid compounds （2-hydroxyl cinnamaldehyde, coumarin, cinnamyl alcohol, cinnamic acid, 2-methoxy cinnamic acid, cinnamaldehyde and 2-methoxy cinnamaldehyde） in Cinnamomi Cortex and Cinnamomi Ramulus. The levels of seven phenylpropanoid compounds in Cinnamomi Cortex and Cinnamomi Ramulus were compared using this method. A total of 48 samples （27 Cinnamomi Cortex and 21 Cinnamomi Ramulus） were purchased in China and analyzed. Quantities of seven phenylpropanoid compounds ranged from 17.5 to 61.6 mg/g in Cinnamomi Cortex and ranged from 9.91 to 23.4 mg/g in Ciunamomi Ramulus. The level of 2-methoxy cinnamic acid in the Cinnamomi Cortex samples was below the LOD, whereas it ranged from 0 to 0.119 mg/g in the Cinnamomi Ramulus samples. The （cinnamyl alcohol＋cinnamic acid）/cinnamaldehyde ratios （R346） of Ciunamomi Cortex and Cinnamomi Ramulus ranged from 0.0121 to 0.0467 and 0.0598 to 0.182, respectively. This ratio could be used to discriminate Cinnamomi Cortex （〈0.05） and Cinnamomi Ramulus （〉0.05）. The extraction rates （Dn） of seven compounds in boiling water were different, with the lowest dissolution for cinnamaldehyde （〈3%） and the highest for cinnamic acid （about 60%）.

Antipyretic Effect of Herba Ephedrae-Ramulus Cinnamomi Herb Pair on Yeast-Induced Pyrexia Rats: A Metabolomics Study

Objective： To investigate the antipyretic mechanism of Herba Ephedrae （Eph）-Ramulus Cinnamomi （RC） herb pair on yeast-induced pyrexia in rats. Methods： Totally 30 qualified male SD rats were randomly assigned to the normal control （NC） group, the pyrexia model （model） group, the Eph, RC and Eph-RC treatment groups by a random digital table, 6 rats in each group. Each rat received a 20% aqueous suspension of yeast （10 mL/kg） except the NC group. The 3 treatment groups were administered 8.1, 5.4 and 13.5 g/kg Eph, RC and Eph-RC respectively at 5 and 12 h after yeast injection, the NC group and the model groups were administered equal volume of distilled water. Rectal temperatures were measured at 0, 6, 8, 10, 12, 15, 18, 24 and 30 h and urine was collected prior to yeast injection and at 6, 10, 18, 24, 30, and 36 h after yeast injection. Then urine metabolomic profiling by gas chromatography tandem mass spectrometry, coupled with multivariate statistical analysis and pattern recognition techniques were used to explore the antipyretic effects of Eph-RC. Partial least squares discriminate analysis was used to analyze the metabolomics dataset including classification and regression in metabolomics plot profiling. Results： Compared with the NC group, rectal temperatures were significantly higher in the model group （P〈0.01）, while 3 treatment groups decreased significantly compared with the model group （P〈0.05 or P〈0.01）. Rectal temperatures of Eph-RC-treated rats started to go down at 6 h, and markedly decreased at 8, 12, 15, 18 and 24 h （P〈0.05 or P〈0.01）, while those of the Eph and RC groups had decreased firstly at 8 h and were markedly lower at 12 h （P〈0.05 or P〈0.01）. Seventeen potential biomarkers related to pyrexia were confirmed and identified, including pyruvic acid, L-phenylalanine, L-tyrosine, phenylacetic acid, hippuric acid, succinic acid, citrate and so on. Eight potential alterations of metabolic pathways including phenylalanine metabolism, citrate cycle, tryptophan metabolism, biosynthesis of valine, leucine and isoleucine, were identified in relation to the antipyretic effects of Eph-RC using MetPA software. Conclusion： The antipyretic effect of Eph-RC herb pair on yeast-induced pyrexia in rats involved correction of perturbed amino acid, fatty acid, and carbohydrate metabolism according to the metabolic pathway analysis with MetPA.

Effects of Tianma(Rhizoma Gastrodiae)and Gouteng(Ramulus Uncariae Rhynchophyllae cum Uncis)on cytochrome P450 enzyme activities in rats

OBJECTIVE:To investigate the efficacy of Tianma(Rhizoma Gastrodiae)and Gouteng(Ramulus Uncariae Rhynchophyllae cum Uncis)on cytochrome P450(CYP450)enzyme activities in rats.METHODS:A cocktail strategy was followed to evaluate the influence of Tianma(Rhizoma Gastrodiae)and Gouteng(Ramulus Uncariae Rhynchophyllae cum Uncis)on the activities of CYP450 isoforms(CYP1A2,CYP3A4,CYP2E1,CYP2C19,CYP2C9,CYP2D6),which were determined by changes in the pharmacokinetic parameters of six probe drugs,theophylline,dapsone,chlorzoxazone,omeprazole,tolbutamide and dextromethorphan.Study groups included,Control group(CG),Tianma(Rhizoma Gastrodiae)group(TM),Gouteng(Ramulus Uncariae Rhynchophyllae cum Uncis)group(GT)and Tianma Gouteng(Gastrodia Uncaria)group(TMGT).RESULTS:No significant differences between Tianma(Rhizoma Gastrodiae)and control groups were found.Compared with the control group,in the Gouteng(Ramulus Uncariae Rhynchophyllae cum Uncis)group both the AUC and t1/2 of dapsone and tolbutamide were reduced,whereas the CL(clearance rate)of dapsone and tolbutamide were increased.Compared with the control group,in the Tianma Gouteng group,the AUC and t1/2 of dapsone and tolbutamide were reduced,the CL of dapsone and tolbutamide were increased,and the AUC and t1/2 of chlorzoxazone were increased and the CL of chlorzoxazone was reduced.CONCLUSION:Tianma(Rhizoma Gastrodiae)has no significant effect on the six CYP450 subtypes.The activities of CYP3A4 and CYP2C9 were increased by Gouteng(Ramulus Uncariae Rhynchophyllae cum Uncis).The activities of CYP3A4 and CYP2C9 were increased,whereas the activity of CYP32E1 was reduced by combined Tianma(Rhizoma Gastrodiae)and Gouteng(Ramulus Uncariae Rhynchophyllae cum Uncis).

Effect of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomi Compatibility on Uric Acid Metabolism and Urinary Neutrophil Gelatinase-Associated Lipocalin and Kidney Injury Molecule-1 in Rats with Hyperuricemia

Objective：To explore the effects of Rhizoma Polygoni Cuspidati and Ramulus Cinnamomicompatibility（PR） on uric acid metabolism and the expression of urinary neutrophil gelatinase-associated lipocalin（NGAL） and kidney injury molecule-1（KIM-1） in rats with hyperuricemia. Methods：Seventy male Sprague Dawley（SD） rats were randomly divided into 7 groups with 10 rats per group, including the normal group, model group, allopurinol group, benzbromarone group and PR groups at 3 doses（3.5, 7, 14 g/kg）. Except the normal group, rats of the other groups were intragastrically administered 100 mg/kg hypoxanthine and 250 mg/kg ethambutol, and subcutaneously injected with 200 mg/kg potassium oxonate. All rats were continuously modeled for 17 days, and gavaged with corresponding drugs. The rats of the normal and model groups were gavaged with saline, once a day, for 2 weeks. The levels of serum uric acid（SUA）, blood urea nitrogen（BUN） and creatinine（Cr） were determined. In addition, the contents of NGAL and KIM-1 in urine and the m RNA and protein expressions of xanthine oxidase（XOD） in liver of hyperuricemia rats were measured by reverse transcription polymerase chain reaction（RT-PCR） and Western blot, respectively. Moreover, the pathological changes of kidney were analyzed by hematoxylin and eosin（HE） stain method. Results：Compared with the normal group, the levels of SUA, BUN, NGAL and KIM-1 and the expressions of hepatic XOD m RNA and protein in the hyperuricemia rats were increased significantly（P〈0.01）. PR significantly decreased the levels of SUA, BUN, NGAL and KIM-1 and down-regulated the m RNA and protein expressions of hepatic XOD（P〈0.05 or P〈0.01）. In addition, the pathological changes of kidney were significantly suppressed by oral administration of PR. Conclusions：PR ameliorated uric acid metabolism and protected renal function, the underlying mechanism was mediated by decreasing the levels of SUA, BUN, NGAL and KIM-1, inhibiting the expression of hepatic XOD and ameliorating the pathological change of kidney.

Preventive effect of different compatibilities of Ramulus Cinnamomai and Radix Paeomiae alba in Guizhi decoction on cardiac sympathetic denervation induced by 6-OHDA

Objective To observe the preventive effect of different compatibilities of Ramulus Cinnamomi(RC)and Radix Paeomiae alba(RPA)in Guizhi Decoction(GZD)on neurotransmitters and their rate-limiting enzymes,and neurotrophic factors of cardiac sympathetic denervation model rats induced by 6-hydroxydopamine(6-OHDA).Methods Totally 54 male Wistar rats were randomly divided into 6 groups,i.e.,the blank control group。

Advances in Research of Epidemic Lung Preventive Mixture

This paper summarizes the Epidemic Lung Preventive Mixture from three aspects of research status, development trends and prospects, so as to deepen the understanding of Epidemic Lung Preventive Mixture and provide a more detailed theoretical basis for its further clinical research and development.

HPLC法测定钩藤提取物中钩藤碱的含量

[目的]测定钩藤提取物中有效成分钩藤碱的含量。[方法]采用HPLC法测定钩藤提取物中钩藤碱的含量,色谱柱为DiamonsilC18(250 mm×4.6 mm,5μm);流动相为无水甲醇-水(60∶40,V/V)并含10 mmol/L三乙胺,冰醋酸调pH 7.5;柱温为30℃;流速为1.0ml/min;紫外检测波长为254 nm。[结果]钩藤碱浓度在11.0～110.0μg/ml范围内与峰面积的线性关系良好(R=0.999 6),平均回收率为96.84%,RSD为1.46%。[结论]该方法简单、快速、准确,可用于钩藤提取物中钩藤碱的含量测定。

钩藤的化学成分研究

[目的]对钩藤的化学成分进行研究。[方法]利用柱层析对钩藤的化学成分进行分离纯化,然后根据化合物的理化性质和光谱数据鉴定所得到化合物的结构。[结果]从钩藤中分离得到了5个化合物,分别鉴定为乌苏酸、β-谷甾醇、β-胡萝卜苷、柯诺辛碱和钩藤碱。[结论]该方法分析研究了钩藤的化学成分,为钩藤的进一步开发和利用提供参考。

麻黄、桂枝及其药对中矿质元素初级形态含量的测定

采用火焰原子吸收分光光度法测定了麻黄(Herbal ephedra)、桂枝(Ramulus cinnamomi)及其药对中Ca、Cr、Fe、Mg、Mn、Zn等6种矿质元素的含量。结果表明,麻黄、桂枝的Ca含量较高,其次是Mg和Fe;Mg的提取率最高,分别为64.74%和45.88%,药对的提取率均介于两种单味药材之间。对这两种药材及其药对中矿质元素初级形态的研究,为进一步探讨矿质元素与药效之间的关系提供参考。

钩藤中钩藤碱的提取和含量测定

[目的]研究钩藤中钩藤碱的提取和含量测定方法。[方法]采用生物碱回流提取方法提取,采用高效液相色谱法对钩藤中钩藤碱的含量进行测定并计算提取率。[结果]专属性考察结果显示,钩藤碱的保留时间约为18.843 min,异钩藤碱的保留时间约在22.323min,两峰分离度良好,且钩藤药材中其他成分不干扰测定;含量测定结果显示,钩藤碱和异钩藤碱的含量分别为23.23、129.17μg/g,加样回收试验平均回收率分别为99.12%、101.33%,RSD分别为1.84%、1.56%。[结论]该方法稳定、有效、准确度高,可用于钩藤中钩藤碱的含量测定。

桂枝抗菌活性成分及其作用机制研究进展

桂枝(Cinnamomi ramulus)是中国中医药经典名方中不可或缺的一味药,主要含挥发油类、有机酸类以及香豆素类等化学成分,其挥发油中的桂皮醛是桂枝的主要有效成分,也是发挥抗菌作用的主要活性成分,桂枝中的其他成分如桂皮酸、香豆素、原儿茶酸、丁香酚、肉桂醇等也具有抗菌活性。通过查阅相关文献,对桂枝的主要抗菌成分、抗菌效果以及作用机制进行了整理和分析,为桂枝的临床应用、新药用价值的挖掘和资源开发利用提供参考。

UFLC‑PDA‑MS/MS Profling of Seven Uncaria Species Integrated with Melatonin/5‑Hydroxytryptamine Receptors Agonistic Assay

Uncariae Ramulus Cum Uncis(Gou-Teng),the dried hook-bearing stems of several Uncaria plants(Rubiaceae),is a wellknown herbal medicine in China.The clinical application of Gou-Teng is bewildered for the morphological and chemical similarity between diferent species.In order to discern their chemical and biological diference,an ultra-fast liquid chromatography equipped with ion trap time-of-fight mass spectrometry(UFLC-IT/TOF-MS)combining with melatonin(MT1 and MT2)and 5-hydroxytryptamine(5-HT1A and 5-HT2C)receptors agonistic assay in vitro was conducted on seven Uncaria species.As a result,57 compounds including 35 indole alkaloids,ten favonoids,fve triterpenoids,fve chlorogenic analogues,and two other compounds were characterized based on their MS/MS patterns and UV absorptions.Specifcally,cadambine-type and corynanthein-type alkaloids were exclusively present in U.rhynchophylla and U.scandens,whereas corynoxine-type alkaloids were commonly detected in all the seven Uncaria plants.Three Uncaria species,U.rhynchophylla,U.macrophylla,and U.yunnanensis showed obviously agnostic activity on four neurotransmitter receptors(MT1,MT2,5-HT1A,and 5-HT_(2C)).This frst-time UFLCMS-IT-TOF analyses integrated with biological assay on seven Uncaria plants will provide scientifc viewpoints for the clinical application of Gou-Teng.

Clinical experience of treating diabetes and its complications with mulberry plants by Chinese medicine master Lv Ren-he

Lv Renhe,a master of Chinese medicine,was familiar with the sources of medicinal materials,the four nature and five flavors,and the characteristics of ascending,descending,floating and sinking.The study of medicine in clinical practice reflects the principle of“using medicine as a soldier”.In the treatment of diabetes mellitus and its complications,Professor Lv made good use of mulberry leaves,mulberry branches,mulberry white bark,mulberry and mulberry parasitism,and gave full play to the modern pharmacological effects of mulberry drugs such as hypoglycemic,hypotensive and lipidemic in the combined use,and achieved satisfactory results.

Investigation on the Mode of Action of the Traditional Chinese Medical Prescription-Yiqihuoxue Formula, an Effective Extravasation Treatment for Cerebral Vascular Microemboli in ApoE-/- mice

Objective: The objective of this study was to investigate the mechanisms underlying anti-embolism and extravasational effects of traditional Chinese medical prescription YiqiHuoxue(YQHX) formula in ApoE-/-mice with cerebral vascular microemboli. Materials and Methods: An ApoE-/-mice model with microemboli was developed by infusing fluorescently labeled heterologous fibrin-rich microparticles into the internal carotid artery of ApoE -/-gene knockout male mice through the common carotid artery. Before microemboli injection, the animals were randomly divided into four groups of 10 animals, treated daily for 6 weeks by intragastric administration: The ApoE-/-control group(physiological saline, 0.2 mL/10 g/d), YQHX group(0.2 ml/10 g/d), clopidogrel group(3 mg/kg/d), and atorvastatin group(3 mg/kg/d);a further group was constituted of normal male C57 BL/6 J mice(with the same genetic background as ApoE-/-mice;normal control group;no treatment;microemboli injection). The mice in each microemboli group were divided into three subgroups, the 2-h, 24-h, and 72-h subgroups, corresponding to the time after microemboli injection. Two hours(or 24 h or 72 h) after microemboli injection, the changes in aortic intima and brain tissue were analyzed by histopathology, the amounts of fluorescent emboli being measured by fluorescence microscopy image analysis. Comparison points included the microemboli induced loss of aorta functions and pathological changes, atherosclerotic plaque, brain ultrastructure and functions, and embolus extravasation. Results: Loss of aorta functions and adverse pathological changes, atherosclerotic plaque, serious damage in brain ultrastructure and functions, and reduced thrombus elimination were obviously serious in microemboli injected ApoE-/-mice. These symptoms were significantly relieved by the YQHX pretreatment:(i) the ratio of thrombus accumulation was increased with a significant decrease in thrombus extravasation in ApoE-/-mice, while YQHX induced an increased thrombus extravasation;(ii) the degree of aortic intimal thickening and brain tissue structural disorders were significantly increased in ApoE-/-mice, but overtly inhibited in the YQHX group;(iii) YQHX restored cell viability and homeostasis in the brain;(iv) YQHX regulated the expression of pro-and anti-inflammatory cytokines in the aorta;and(v) YQHX reduced cortical nerve nuclei pyknosis, edema, liquefaction, and necrosis induced by brain hypoxia, especially in the 24 h and 72 h groups. Conclusions: These findings indicate that the protective effects of YQHX on the brain against microemboli-induced injury may be attributed to the activation of extravasation mechanisms, which are involved in the cerebrovascular injury pathway and constitutively important in the progression of ischemic stroke.