Sodium Aescinate Alleviates Neuropathic Pain in Rats by Suppressing the TLR4/NF KB Pathway Activation after Paclitaxel Chemotherapy

Background: Emerging evidence suggests that chemotherapy-induced peripheral neuropathy (CIPN) is a significant side effect of chemotherapeutic drugs. Many experiments have proved that sodium aescinate (SA) has definite pharmacological effects such as anti-infection, anti-exudation, anti-edema, anti-tumor as well as neuroprotection, and the drug side effects are mild. However, no study has explored whether SA is involved in the analgesic effect of paclitaxel (PAC) induced neuropathic pain in rats. Methods: Rats were given an intraperitoneal injection of PAC (2.5 mg/Kg intraperitoneally on days 1, 3, 5, and 7), while SA 25 mg/kg intraperitoneally was administered daily for 14 consecutive days. The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) of rats were examined on experimental days 3, 5, 7, 11, 14. All rats were sacrificed on day 15 of the experiment, and L4-6 spinal cords were removed. Subsequently, immunohistochemistry, HE staining, ELISA, RT-qPCR, Western blotting were applied to evaluate cytoskeletal protein expression (NF-L and NF-M), spinal nerve structural integrity, proinflammatory factor contents (TNF-α, IL-1β, and IL-6), and protein content of the TLR4/NF-κB pathway, respectively. Results: After the rats developed PAC induced pain behaviors, multiple injections of SA rendered the rats with elevated MWT and TWL values, decreased expression of NF-L and NF-M in the spinal cord, materially downregulated content of proinflammatory factors, and reduced amounts of TLR4 and p-NF-κB protein levels. Conclusions: The results of the present study preliminarily indicate that SA has an analgesic effect on rats with CIPN induced by PAC injection, and the mechanism may be related to blocking the TLR4/NF-κB signaling pathway, inhibiting the expression of proinflammatory factors, and alleviating cytoskeletal disorders.

CARMA3/NF-κB signaling contributes to tumorigenesis of hepatocellular carcinoma and is inhibited by sodium aescinate

BACKGROUND Primary hepatocellular carcinoma (HCC) is a very malignant tumor in the world. CARMA3 plays an oncogenic role in the pathogenesis of various tumors. However, the function of CARMA3 in HCC has not been fully clarified. AIM To study the biological function of CAEMA3 in HCC. METHODS Tissue microarray slides including tissues form 100 HCC patients were applied to access the expression of CARMA3 in HCC and its clinical relevance. Knockdown and overexpression of CARMA3 were conducted with plasmid transfection. MTT, colony formation, and apoptosis assays were performed to check the biological activity of cells. RESULTS Higher expression of CARMA3 in HCC was relevant to poor prognostic survival (P < 0.05). Down-regulation of CARMA3 inhibited proliferation and colony formation and induced apoptosis in HCC cell lines, while increasing its expression promoted tumorigenesis. We also found that sodium aescinate (SA), a natural herb extract, exerted anti-proliferation effects in HCC cells by suppressing the CARMA3/nuclear factor kappa-B (NF-κB) pathway. CONCLUSION Overexpression of CARMA3 in HCC tissues correlates with a poor prognosis in HCC patients. CARMA3 acts pro-tumorigenic effects partly through activation of CARMA3/NF-κB. SA inhibits HCC growth by targeting CARMA3/NF-κB.

Effect of β-sodium aescinate on hypoxia-inducible factor-1α expression in rat brain cortex after cardiopulmonary resuscitation

BACKGROUND:This study was undertaken to investigate the expression of hypoxia-inducible factor-1a(HIF-1a) in rat cerebral cortex and the effects of p-sodium aescinate(SA) administration after return of spontaneous circulation(ROSC).METHODS:Sixty rats were divided into three groups:SA group,injected intraperitoneally with SA instantly after ROSC;control group,injected intraperitoneally with normal saline;and shamoperated group,without cardiac arrest or SA.The cardiac arrest model was established using asphyxiation and intravenous potassium chloride.Blood was sampled 1,6,12,and 24 hours after ROSC.Protein and mRNA levels of HIF-1a,VEGF and EPO were detected in the cerebral cortex by immunohistochemistry and real-time RT-PCR;serum levels of NSE and S100 P were determined by enzyme-linked immunosorbent assays.RESULTS:Serum S100β and NSE were significantly increased in the control group versus the sham-operated group 1,6,12 and 24 hours after ROSC(P<0.05).Protein and mRNA levels of HIF-1a,VEGF and EPO were significantly increased in the control rats(P<0.05).Serum NSE and S100 P were significantly decreased in the SA group versus the control group 1,6,12 and 24 hours after ROSC(P<0.05).Protein and mRNA levels of HIF-1a,VEGF and EPO were significantly increased in the SA group(P<0.05).CONCLUSIONS:The expression of HIF-1a is increased in rat cerebral cortex after ROSC,and SA up-regulates the expression of HIF-1α.The up-regulation of HIF-1α improves the resistance of the cortex to ischemia and hypoxia and contributes to neuroprotection,possibly because of up-regulation of EPO and VEGF expression.

Anti-proliferation and apoptosis-inducing effects of sodium aescinate on retinoblastoma Y79 cells

AIM:To investigate the anti-proliferation and apoptosisinducing effects of sodium aescinate(SA)on retinoblastoma Y79 cells and its mechanism.METHODS:Y79 cells were cultured at different drug concentrations for different periods of time(24,48,and 72 h).The inhibitory effect of SA on proliferation of Y79 cells was detected by the cell counting kit-8(CCK-8)assay,and the morphology of Y79 cells in each group was observed under an inverted microscope.An IC50 of 48 h was selected for subsequent experiments.After pretreatment with SA for 24 and 48 h,cellular DNA distribution and apoptosis were detected by flow cytometry.Real-time qunatitative polymerase chain reaction(RT-qPCR)and Western blot were used to assess changes in related genes(CDK1,CyclinB1,Bax,Bcl-2,caspase-9,caspase-8,and caspase-3).RESULTS:SA inhibited proliferation and induced apoptosis of Y79 cells in a time-dependent and concentrationdependent manner.Following its intervention in the cell cycle pathway,SA can inhibit the expression of CDK1 and Cyclin B1 at the mRNA and protein levels,and block cells in the G2/M phase.In caspase-related apoptotic pathways,up-regulation of Bax and down-regulation of Bcl-2 caused caspase-9 to self-cleave and further activate caspase-3.What’s more,the caspase-8-mediated extrinsic apoptosis pathway was activated,and the activated caspase-8 was released into the cytoplasm to activate caspase-3,which as a member of the downstream apoptotic effect group,initiates a caspase-cascade reaction that induces cell apoptosis.CONCLUSION:SA inhibits the proliferation of Y79 cells by arresting the cell cycle at the G2/M phase,and induces apoptosis via the caspase-related apoptosis pathway,indicating that SA may have promising potential as a chemotherapeutic drug.

Influence of compound aescine gel on ultrastructure of vein infused mannitol and its mechanism

Purpose: When hypertonic solution 20% mannitol solution was injected into vein, inflammatory mediators and Mitogen-activated protein kinases activated by mannitol can directly induce the fading of vascular endothelial cell, which leads to phlebitis. The study aims to observe the influences of reparil-gel N coated at the proximal parts of the puncture point and basing on this along with infusing heated mannitol to veins to the injure and ultrastructure of veins which were infused the 20% mannitol solution by indwelling needle in vein. Methodology: There are 15 adult New Zealand rabbits. We randomly divided 24 ear veins of 12 adult New Zealand rabbits into Control group, Gelatum group, Gel heated group and injected 20% mannitol solution by vein detained needle in three groups. In Gelatum group, we coated the proximal end of the puncture point with a thin layer of compound aescine gel. Based on Gelatum group, we heated 20% mannitol solution to 35oC-37oC in Gel heated group. Then we observed the intravenous parts and took the veins of each group out to observe their morphology and ultrastructural after the second day of transfusion. 6 ear veins of the rest 3 rabbits as Health group weren’t given any interventions, the veins were taken out to observe their morphology. Results: Comparison between Gelatum group and Gelatum heated group on vascular dilatation, Infiltration of inflammatory cell and Formation of thrombus had no significance, P> 0.05, while the case was different for the comparison on injury of vascular wall, perivascular edema and perivascular hemorrhage, P< 0.05). The statistical significance exists between control group and Gelatum group and Gel heated group, P< 0.05. It was observed under the electron microscope that, in control group, the membrane of endothelial cell peeled off and the mitochondria swelled and vacuolized. In Gelatum group, the membrane of endothelial cell was defective, the parts of the mitochondria were fuzzy, intercellular substance was slightly edematous. In Gel heated group, the mitochondria were clear and intercellular substance slightly swelled. It could be found that the function of phagocyte was complete. Conclusions: Compound aescine gel can prevent phlebitis or reduce the incidence of phlebitis. The combined intervention of coating with a thin layer of compound aescine gel and heating mannitol solution can produce better effect.

The study on the mechanism of Sodium aescinate combined with Xuesaitong for cerebral thrombosis treatment

Objective:To observe the mechanism of Sodium aescinate combined with Xuesaitong for cerebral thrombosis treatment and offer clinical help to cerebral thrombosis treatment. Methods:A total of 92 patients with cerebral thrombosis were selected and randomly divided into groups: the observation group (46 people) and the control group (46 people).The patient in the control group were treated with conventional therapy and the patients in the observation group were treated with Sodium aescinate combined with Xuesaitong on the basis of conventional therapy. Inflammatory factors (IL-6, IL-10, TNF-α and CRP), Nerve cell factor (NSE, NGF and NTF) and blood rheology indexes (Fg, BVH, BVL and PV) were detected and analyzed before and after treatment.Results:The comparison of inflammatory factors, Nerve cell factor and blood rheology indexes in the two groups before treatment were not statistically significant. Inflammatory factors (IL-6, TNF-α and CRP), Nerve cell factor (NSE) and blood rheology indexes (Fg, BVH, BVL and PV) in both groups after treatment significantly decreased compared with that before treatment. Inflammatory factors (IL-10), Nerve cell factor (GF and NTF) in both groups after treatment significantly increased compared with that before treatment. The changes were statistically significant. Inflammatory factors (IL-6, TNF-α and CRP), Nerve cell factor (NSE) and blood rheology indexes (Fg, BVH, BVL and PV) in observation group after treatment decreased more significantly than that in control group, and inflammatory factors (IL-10), Nerve cell factor (GF and NTF) increased more significantly than that in control group. The difference between two groups was considered statistically significant.Conclusion:Sodium aescinate combined with Xuesaitong could inhibit the inflammatory reaction, improve the blood flow condition and promote rehabilitation in patients with cerebral thrombosis. So it has a very important clinical significance of the treatment to cerebral hemorrhage.

Intervening Effect of Sodium Aescinate on Pulmonary Fibrosis in Rats with Acute Lung Injury

[Objectives] The aim was to investigate the intervening effect of sodium aescinate on pulmonary fibrosis in rats with acute lung injury( ALI). [Methods] The rats were randomly divided into normal group,model group and sodium aescinate group. The rat model of ALI was induced by administration of oleic acid. The rats in the sodium aescinate group were intravenously injected with sodium aescinate according to the amount of 4 mg/kg for 14 consecutive d. Then,11 rats were selected randomly from each group and slaughtered on Day 1 and Day 14,respectively after last administration. The body mass index,arterial partial pressure of oxygen( PaO_2),oxygenation index( PaO_2/FiO_2),lung index,wet/dry mass ratio of lung,serum IL-1β,TNF-α,PC Ⅲ and TGF-β1 levels of the rats were analyzed. [Results]No significant differences were found in body mass index,lung index or lung wet/dry mass ratio among different groups. Compared with the model group,the PaO_2 and PaO_2/FiO_2 ratio increased significantly( P < 0. 05),the serum IL-1β,TNF-α,PC Ⅲ and TGF-β1 levels declined significantly( P <0. 05),the lung histopathological damage was reduced,and the semi-quantitative histological score( IQA) of damaged lung tissue decreased significantly( P < 0. 01). [Conclusions]Sodium aescinate can reduce the levels of inflammatory factors in rats with ALI,with certain intervening on pulmonary effect.

A Rapid and Simple Quantitative Method for the Active Ingredients of Aescin in the Extraction Process Using Near Infrared Spectroscopy

To achieve a rapid and simple detection for the active ingredients of Aescin in the extraction process using near-infrared spectroscopy (NIR) and to realize the state monitoring and quality control of the extraction process. Partial least square regression (PLS) was applied to build the near-infrared calibration models, and the applicability of the model was investigated by predicting the unknown samples in the extraction process. The correlation coefficients of the established Aescin models (A, B, C, D) were 0.9836, 0.9831, 0.9833, 0.9824, and the prediction standard deviations (SEP) were 0.05636, 0.05043, 0.02412, 0.05636, respectively. This study suggests that the proposed model has superior stability and accuracy. NIR spectroscopy technique provides a novel efficient and environmentally friendly approach to the rapid determination of four Aescin key quality indicators (A, B, C, D) in the extraction, which was solved the problem that the lack of state monitoring during the extraction of Aescin, thereby improved the quality of Aescin.

Mechanism of Sodium aescinate combined with Torasemide for cerebral hemorrhage treatment

Objective:To observe the mechanism of Sodium aescinate combined with Torasemide for cerebral hemorrhage treatment and offer clinical help to cerebral hemorrhage treatment. Methods:86 patients with cerebral hemorrhage were selected and randomly divided into groups: the observation group (43 people) and the control group (43 people). The patient in the control group were treated with conventional therapy and the patients in the observation group were treated with Sodium aescinate combined with Torasemide on the basis of conventional therapy. Oxidative stress indexes [Glutathione peroxidase (GSH-Px), catalase (CAT), malondialdehyde (MDA), superoxide dismutase (SOD)], serum inflammatory factors [interleukin-6 (IL-6), interleukin-2 (IL-2), C reactive protein (CRP) and tumor necrosis factor-α (TNF-α)] and hemorheology indexes [Hematokrit (PCV), plasma viscosity (PV), erythrocyte aggregation index (PAdT) and whole blood viscosity at high shear rate (WHV)] were detected and analyzed before and after treatment.Results:The comparison of oxidative stress indexes, serum inflammatory factors and hemorheology indexes in the two groups before treatment were not statistically significant (P>0.05). Serum inflammatory factors (TNF-α, IL-6, CRP), MDA and hemorheology indexes (WHV, PV, PCV and PAdT) in both groups after treatment significantly decreased compared with that before treatment (P<0.05);Oxidative stress index (GSH-Px, CAT, SOD and MDA) and IL-2 in both groups after treatment significantly increased compared with that before treatment (P<0.05). Serum inflammatory factors (TNF-α, IL-6, CRP), MDA and hemorheology indexes (WHV, PV, PCV and PAdT) in observation group after treatment decreased more significantly than that in control group (P<0.05), and oxidative stress index (GSH-Px, CAT, SOD and MDA) and IL-2 increased more significantly than that in control group (P<0.05).Conclusions:Sodium aescinate combined with Torasemide could decrease the oxidative stress and inflammatory reaction in patients with cerebral hemorrhage, which is beneficial to improving the condition of circulating blood stasis, accelerating microcirculation. So it has a very important clinical significance of the treatment to cerebral hemorrhage.

Protective Effects of the Preconditioning with Different Doses of Sodium Aescinate on Tourniquet-induced Ischemic Reperfusion

Objective：To evaluate the protective effects of sodium aescinate（SA）preconditioning on the tourniquet-induced ischemia-reperfusion（l/R）injury after limbs operation.Methods：Seventy-five patients with gradeⅠ-Ⅱissued by American Society of Anesthesiology undergoing lower limb operation were randomly assigned to 3 groups：the control group,low-dose SA-treated group and high-dose SA-treated group;each group enrolled 25 patients.The patients were treated with 5 mg and 10 mg SA 30 min before tourniquet inflation in the two treatment groups separately,while the patients in the control group received normal saline.Venous blood samples were obtained before tourniquet was inflated（T0 baseline）.And 5（T1）,10（T2）,20（T3）min after tourniquet was released.The nitric oxide（NO）,malondialdehyde（MDA）and superoxide dismutase（SOD）levels were determined by commercial kits.Meanwhile,arterial pressure（MAP）and heart rate（HR）were monitored from an automatic invigilator.Results：In the control group,MDA and NO levels were increased,and SOD and MAP were decreased significantly after tourniquet deflation compared to TO baseline（P0.05）.After tourniquet deflation,MDA and NO levels in the two treated groups were significantly decreased;meanwhile,SOD levels and MAP were increased,and the variations of HR were more stable compared with the control group（all P0.05）.There was no significant difference in all of the above between the two treated groups（P0.05）. Conclusion：The protective effects of SA preconditioning on tourniquet-induced limb l/R injury might possibly contribute to the increasing of SOD levels,and MAP and the decreasing of MDA and NO levels.