Ginkgolide K inhibited neurovascular unit injury against oxygen-glucose deprivation and reperfusion by regulating hypoxia-inducible factor-1α pathway / 中国药理学通报

Aim To investigate the protective effects of ginkgolide K (GK) on neurovascular unit injured by ischemic stroke and the potential mechanism associated with hypoxia-inducible factor-1α (HIF-1α) pathway. Methods The BV-2 cells and EA. hy926 cells suffered from oxygen-glucose deprivation and reperfusion (OGD/R) were applied to mimic the injury of neurovascular unit induced by cerebral ischemia in vitro. After 4 h OGD insult, BV-2 cells and EA. hy926 cells received reperfusion and treated with GK. The levels of inflammatory cytokines in the supernatant of BV-2 cells were detected, while the protective effects of GK on EA. hy926 cells were also evaluated after GK administration for 24 h. The p-Akt and p-Erk expressions were examined by Western blot after 1 h of GK treatment, while HIF-1α was detected after 6 h of GK treatment. In addition, PI3K inhibitor LY294002 was applied to further verify the potential mechanisms underlying the beneficial effects of GK. The expressions of p-Akt after 1 h of GK treatment, and the protein levels of HIF-1α pathway after 6 h of GK treatment were also analyzed by Western blot. Results GK significantly inhibited the levels of TNF-α, IL-6 and IL-1β in supernatant of BV-2 cells injured by OGD/R, through increasing p-Akt and decreasing p-Erk expressions, and then affecting HIF-1α pathway. In addition, LY294002 reduced the regulatory effect of GK. Furthermore, GK significantly improved viability and inhibited the release of LDH in supernatant of EA. hy926 cells suffered from OGD/R, and up-regulated the expressions of p-Akt, HIF-1α, HO-1 and VEGF, while cleaved caspase-3/9 was inhibited. Conclusions GK exerts multi-effects on reducing neurovascular unit injury induced by ischemic stroke, and the potential mechanism might be associated with the different regulatory effects of HIF-1α in different cells.

Molecular mechanism of Epimedii Folium in treatment of osteoporosis based on network pharmacology / 中国中药杂志

Osteoporosis is a systematic bone disease,characterized by deterioration in bone mass or micro-architecture,and increasing risk of fragility and fractures. With the development of aging problems,osteoporosis has been a global health problem. At present,due to the undesirable side effects of synthetic osteoporosis inhibitors,more efforts are made in treatment of osteoporosis by traditional Chinese medicine and its prescriptions. Epimedii Folium,one of the most common herbs for osteoporosis,has attracted great attentions worldwide.In this study,network pharmacology was employed to analyze the active components and potential molecular mechanism of Epimedii Folium on osteoporosis. Component-target network analysis showed that those with higher molecular network degree were flavonoids,with estrogen-like activity,antioxidation and free radical-scavenging activities,playing certain roles in preventing and treating osteoporosis. On the other hand,the targets with high degree were mostly related with sex hormone,osteoclast differentiation,bone matrix degradation,and reactive oxygen species in drug-target network. Multiple components of Epimedii Folium could be interacted with these targets. This study shows that Epimedium could prevent and treat osteoporosis through multiple active ingredients acting on multiple targets.

Microenvironmental cues influence the reprogramming of somatic cells to induced pluripotent stem cells / 中国组织工程研究

BACKGROUND: Induced pluripotent stem cells hold enormous potential as a tool to generate cells for disease therapy and modeling. However, the lower efficiency of cell reprogramming using exogenous transcription factors limits the clinical use of induced pluripotent stem cells. Recent studies have indicated that microenvironmental cues can promote the production of pluripotent cells. Further summarization on these reports will provide new strategies to enhance the safety and efficiency of the reprogramming technology. OBJECTIVE: To summarize the effect of microenvironmental cues on the reprogramming of somatic cells to induced pluripotent stem cells and the maintenance of pluripotency. METHODS: Scientific papers from 2006 to present included in Web of Science were searched by the first author. Eighty papers focusing on the effect of microenvironmental cues on the reprogramming of somatic cells to induced pluripotent stem cells and the maintenance of pluripotency were included. RESULTS AND CONCLUSION: Cell-matrix interaction and cell-cell adhesion regulate gene expression and have effects on the epigenetic states of the chromatin through a direct connection between actin cytoskeletons and the nuclear membrane; thus, manipulation of the cell-matrix interaction and cell-cell adhesion can improve the induction of pluripotency. In addition, other environmental factors including hypoxia, dynamic culture and electrical stimulation can also promote cell reprogramming.

Effect of compatibility of ginkgolide A, ginkgolide B and ginkgolide K / 中国中药杂志

To investigate the best active compatibility of ginkgolide A, B and K (GA，GB，GK). The effects of GA, GB, GK alone, combinations of each two of them, and combinations of these three components on platelet-activating factor (PAF)-induced platelet aggregation activity and rat cerebral ischemia reperfusion model (tMCAO) were compared in this study. Different compatibilities of GA, GB and GK could significantly reduce the maximum aggregation rate of PAF-induced platelet aggregation, and the effect was most obvious in combination of the three. Different compatibilities of GA, GB and GK could alleviate the neural function, cerebral infarction volume and cerebral edema in the tMCAO model of rats to different degrees, and the effect of combinations of the three was stronger than those of combinations of two and single use. The combination of all of GA, GB and GK had the strongest effect on nerve injury caused by anti-platelet aggregation in tMCAO rats.

Antagonistic effect of ginkgolide homologues on PAF-induced platelet aggregation and neuroprotective effect / 中国中药杂志

To study the antagonistic effect of ginkgolide homologues on platelet-activating factor (PAF)-induced platelet aggregation and investigate its neuroprotective effect. PAF was used as a coagulant, and ginkgolides were added to the rabbit blood samples respectively. The inhibitory effect of each compound on platelet aggregation was detected by turbidimetry. In L-glutamate induced primary cortical neuron cell injury model, MTT assay was used to detect cell viability. Intracellular free Ca2+ concentration in neurons was measured by using the fluorescent Ca2+ indicator Fura-2 AM. Morphological observation and Hoechst 33258 staining were used to detect the inhibitory effect of ginkgolide on neuronal apoptosis. The results showed that the inhibitory effect on PAF-induced platelet aggregation activity in ginkgolide homologues was ginkgolide K (GK), ginkgolide B (GB), ginkgolide A (GA), ginkgolide C (GC), ginkgolide M (GM), ginkgolide J (GJ) and ginkgolide (GL) from high to low. GB and GK (1-100 μmol•L ⁻¹) could significantly reduce the cell damage caused by L-glutamate, with survival rate increasing, intracellular calcium concentration reducing and cell morphology restoring. This paper has identified the activities and characteristics of various compounds of ginkgolide homologues on PAF-induced platelet aggregation as well as its neuroprotective effect.

Effects of ginkgolide A, B and K on platelet aggregation / 中国中药杂志

To investigate the effects of ginkgolide A (GA), ginkgolide B (GB) and ginkgolide K (GK) on platelet aggregation in rabbits, and compare the similarities and differences among these three components. The effects of different doses of ginkgolide A, B and K on platelet aggregation induced by platelet activating factor (PAF) were observed by using in vitro experiment. The results showed that three compounds could inhibit platelet aggregation induced by PAF in vitro, and the intensity was GK> GB> GA. It was further found that all of them can mobilize [Ca2+]i and enhance intracellular c-AMP level in a dose-dependent manner, which was consistent to the ability to antagonize PAF receptor. These findings indicated that GK was highly selective for PAF receptor, and may inhibit platelet aggregation by activating cAMP signaling pathway and inhibiting intracellular [Ca2+]i mobilization; GB and GA also had strong antagonism to PAF receptor, but the effect was weaker than that of GK.

Effects of ginkgolide K on platelet aggregation activity and neuroprotection / 中国中药杂志

To investigate the antagonism effects of different concentrations of ginkgolide K(GK) on platelet activating factor (PAF)-induced platelet aggregation and neuroprotective effect on cells and animal models of ischemia-reperfusion injury. GK-containing serum in rabbit was prepared, and the effects of GK-containing serum on PAF-induced platelet aggregation was observed by platelet aggregation assay. The effect of different concentrations of GK on apoptosis of SH-SY5Y cells injured by oxygen-glucose deprivation/reoxygenation (OGD/R) was investigated by Hoechst 33342/PI double staining in OGD/R cell model. The focal cerebral ischemia-reperfusion model (I/R)was established in rats to detect the effects of GK on neurobehavioral scores and cerebral infarction volume. GK could inhibit PAF-induced platelet aggregation, reverse the apoptosis induced by OGD/R injury and improve the neurobehavioral score and cerebral infarction volume after cerebral ischemia-reperfusion injury in rats in a dose-dependent manner. GK can inhibit PAF-induced platelet aggregation and improve nerve injury after cerebral ischemia-reperfusion.

Protective effects of Ginkgo Terpene Lactones Meglumine Injection on focal cerebral ischemia in rats / 中国中药杂志

To investigate the protective effects of ginkgo diterpene lactone meglumine injection (GDLMI) on cerebral focal ischemia reperfusion injury induced by middle cerebral artery occlusion (MCAO) in rats, and explore its possible mechanism. One hundred and forty male SD rats were randomly divided into sham operation group, model group, ginkgo biloba extract injection (Ginaton, 1.0 mL•kg⁻¹) group, nimodipine (0.4 mg•kg⁻¹) group, and GDLMI (5.2, 2.6, 1.3 mg•kg⁻¹) groups; All of rats received corresponding drugs by tail vein injection 4 days before operation (normal saline in model group and sham operation group). Except the sham operation group, the cerebral ischemic stroke model was established by MCAO method in right brain of the other rats. After 3 h of ischemia, all the animals received intravenous administration again. The neurobehavioral scores of rats after ischemia-reperfusion were evaluated and the infarct rate of brain tissue was observed by TTC staining. The super oxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA) and lactic acid (LA) contents in brain tissue homogenate and the concentration of Ca2+, glutamate (Glu) and aspartate (Asp), creatine phosphate kinase (CK-BB) and lactate dehydrogenase (LDH) content changes in cerebrospinal fluid were measured. As compared with the sham operation group, the cerebral infarction rate was increased significantly in the model group; the content of MDA and LA in the homogenate of brain tissue was increased, and the content of GSH and SOD was decreased; in cerebrospinal fluid, Ca2+ concentration was decreased, and the content of Glu and Asp, CK-BB and LDH increased significantly. As compared with the model group, the high and medium dose GDLMI groups can significantly reduce the cerebral infarction rate and improve the symptoms of neurological impairment; increase SOD and GSH activity, reduce MDA and LA content in serum; increase Ca2+ concentration in cerebrospinal fluid and decrease the content of neurotransmitter Glu and Asp as well as CK-BB and LDH. GDLMI could obviously improve neurologic impairment in model rats, and the mechanism may be related to recovering the blood brain barrier, scavenging free radicals, decreasing free Ca2+ inflow into the cells and the content of excitatory amino acid in cerebrospinal fluid to improve its protective effect on cerebral ischemia.

Cerebral vascular protective effect of ginkgo diterpene lactone meglumine injection / 中国中药杂志

The left middle cerebral artery occlusion (MCAO) model was made by inserting the nylon thread plug into the internal carotid artery. The behavioral score, cerebral infarction area, brain water content, ethidium bromide (EB) spillover, coagulation four indices, occludin and MMP-9 expression in brain tissues were detected after 14 days of administration, to investigate whether the protective effect of ginkgo diterpene lactone meglumine injection (GDLMI) which had obvious protective effect on cerebral ischemic injury in the previous experiment was related to reducing the permeability of the blood-brain barrier (BBB) and reducing the risk of bleeding, and to explore its possible mechanism of action. The results showed that GDLMI could effectively alleviate the behavioral changes caused by MCAO at 24 h, reduce the behavioral score, improve the edema of brain tissue, reduce the EB overflow rate, reduce the bleeding tendency caused by long-term administration, significantly reduce the occlusion deficiency in ischemic brain tissue of model rats, and down-regulate MMP-9 expression. The above results indicate that GDLMI has obvious effect on cerebral ischemia, and the therapeutic effect of GDLMI may mainly depend on lowering the permeability of blood-brain barrier to improve brain edema.

Antiviral effects of Reduning injection against Enterovirus 71 and possible mechanisms of action / 中国天然药物

The present study was designed to evaluate the protective effects of Reduning injection against Enterovirus 71 (EV71) in Vero cells and in mice. The Vero cells were infected with 100 and 50 TCID50 (50% tissue culture infective dose) of EV71, respectively. The inhibition of Reduning injection on cytopathic effect (CPE) was detected. Meanwhile, a mouse model produced by intraperitoneal EV71-infection (10(6) TCID50), was used to investigate the protective effects of Reduning injection. The total survival rate, living time, daily survival rate, weight ratio, and score for symptoms were examined. The viral loads in Vero cells and muscle tissues were detected using real-time PCR. Finally, the content of cytokines was analyzed by ELISA. In the Vero cells, 2.5 mg crude drug·mL(-1) of Reduning injection inhibited CPE induced by EV71 infection. In the mice, 1.3 g crude drug·kg(-1) of Reduning injection rescued death triggered by infection, in comparison with model group. Moreover, the survival rate, weight ratio, and clinical scores were also improved. The viral RNA copies in the Vero cells and the mice muscle tissues were reduced. Besides, the steep EV71-induced accumulations of TNF-α and MCP-1 were decreased by Reduning injection. In conclusion, Reduning injection showed promising protective effects against EV71 in Vero cells and in mice.

Study on contribution of main components in Guizhi Fuling capsule based on molecular imprinting technique and activity screening / 中国中药杂志

To clarify the active components in Guizhi Fuling capsule in treatment of intrinsic dysmenorrhea, pelvic inflammation and hysteromyoma, main components were gradually knocked out from the capsules, the effects of knockout capsules on uterine contraction, TNF-α secretion, murine splenocytes (SPL) and hysteromyoma cells proliferation were evaluated, respectively. The inhibition of capsules on uterine contraction was weakened by gradient knockout of paeoniflorin, paeonol, and amygdalin. The suppression of capsulte on TNF-α secretion was reduced by gradient knockout of gallic acid, cinnamaldehyde, pentagalloylglucose, and pachyman. The promotion of SPL cells proliferation was reversed by gradient knockout of gallic acid, paeoniflorin, cinnamaldehyde, quercetin, and pachyman. The depression of capsules on hysteromyoma cells proliferation was attenuated by gradient knockout of paeoniflorin, paeonol, pentagalloylglucose, and albiflorin. In conclusion, the compounds mentioned-above could be the key active basis of Guizhi Fuling capsule in treatment of intrinsic dysmenorrhea, pelvic inflammation and hysteromyoma.

Study of effective components and molecular mechanism for Guizhi Fuling formula treatment of dysmenorrhea, pelvic inflammatory disease and uterine fibroids / 中国中药杂志

In this study, the active components and potential molecular .mechanism of Guizhi Fuling formula in treatment on dysmenorrhea, pelvic inflammation, and hysteromyoma were investigated using network pharmacological methods. Sterols and pentacyclic triterpenes, with high moleculal network degree, revealed promising effects on anti-inflammatory, analgesic, anti-tumor, and immune-regulation, according to D-T network analysis. On the other hand, the targets with high degree were involved in inflammatory, coagulation, angiopoiesis, smooth muscle contraction, and cell reproduction, which showed the novel function in anti-dysmenorrhea, pelvic inflammation, and hysteromyoma. Furthermore, the formula was indicated to play a key role in smooth muscle proliferation, inhibition of new vessels, circulation improvement, reduction of hormone secretion, alleviation of smooth muscle, block of arachidonic acid metabolism, and inflammation in uterus. Thus, the main mechanism of Guizhi Fuling formula was summarized. In conclusion, Guizhi Fuling formula was proven to alleviated dysmenorrhea, pelvic inflammation, and hysteromyoma by acting on multiple targets through several bioactive compounds, regulating 21 biological pathways.

Antiviral effect of Reduning Injection against coxsackievirus A16 / 中草药

Objective: To verify the antiviral effects of Reduning Injection against coxsackievirus A16 (CoxA16) in vivo and in vitro. Methods: Vero cells and 5-day-old suckling mice, injected with 75/50 and 106 TCID50 CoxA16, were used as evaluation models. The preventive influences of Reduning Injection against CoxA16 in Vero cells were assessed in the models. The effects of Reduning Injection on the mortality, survival time, change rate of body weight, and clinical symptom scores of suckling mice were observed. Results: In Vero cells, cytopathic effects induced by 75 and 50 TCID50 CoxA16 were obviously relieved by crude drug5.0 mg/mL Reduing Injection respectively. Meanwhile, mice death caused by CoxA16 was markedly rescued, survival time was prolonged, and growth inhibition was recovered by Reduing Injection. Meanwhile, the clinical symptom induced by virus was also improved. Conclusion: The endogenous and exogenous antivirus effects of Reduning Injection on CoxA16 are proven.