We investigated whether Panax notoginseng saponin (PNS-R1) attenuates allergic rhinitis (AR) through AMPK/Drp1-mediated mitochondrial fission. AR model was established in mice by Ovalbumin (OVA). In vitro, human nasal epithelial cells (HNEpCs) were stimulated using recombinant human interleukin 13 (IL-13). PNS-R1 was administrated in vivo and in vitro. Then, HE staining of nasal tissue, ELISA detection of immunoglobulin E (IgE) and proinflammatory cytokine levels in serum and nasal lavage fluid, flow cytometry analysis of Th1/Th2 ratio and apoptosis, TUNEL staining, Western blot, detection of reactive oxygen species (ROS) and mitochondrial ROS, immunofluorescence analysis of Tom20 and mitochondrial fission protein Drp1 co-localization, and mitochondrial membrane potential detection, were performed. PNS-R1 attenuated allergic symptoms in AR mice, decreased OVA-specific IgE, IL-4, IL-6, IL-8, IL-13, and TNF-α levels, and restored the Th1/Th2 imbalance. Meanwhile, we found that PNS-R1 treatment significantly reduced apoptosis, ROS production, and co-localization of Tom20 and Drp1 in the nasal epithelium of AR mice. In vitro, we found that PNS-R1 upregulated mitochondrial membrane potential and reduced ROS and mitochondrial ROS production as well as Cleaved-caspase-3/9, Bax, Cyt-c, Apaf-1 expression and mitochondrial fission. Mechanistically, we found that PNS-R1 downregulated Drp1 phosphorylation (Ser 616) and Drp1 translocation in an AMPK-dependent manner, promoted MFN2 expression, and reduced TXNIP, NLRP3, Caspase-1, and IL-1β expression. PNS-R1 may protect mitochondrial integrity by inhibiting AMPK/Drp1 and TXNIP/NLRP3 signaling pathway, thereby alleviating AR symptoms in mice. PNS-R1 may have great potential as a therapeutic agent for AR.
目的 研究玫瑰花总黄酮(total flavonoids from Rosa rugosa,TFR)对大鼠脑缺血/再灌注损伤(cerebral ischemia reperfusion injury,CIRI)的影响,探讨TFR是否通过磷脂酰肌醇-3-激酶/蛋白激酶B(phosphoinositide 3-kinase/protein ki-nase B,PI3K/AKT)信号通路和内质网应激(endoplasmic re-ticulum stress,ERS)途径调控神经细胞凋亡.方法 将SD大鼠随机分成假手术组、模型组、TFR低、中、高剂量组(50、100、200 mg·kg-1·d-1)组,灌胃7 d,末次给药1 h后线栓法制备大脑中动脉阻塞/再灌注(middle cerebral artery occlu-sion/reperfusion,MCAO/R)模型.24 h后检测大鼠神经行为学变化、脑梗死面积、脑组织含水量;HE和尼氏染色观察病理相关指标;TUNEL染色观察神经细胞凋亡情况;Western blot检测Bcl-2、Bax、cleaved Caspase-3、PI3K、p-PI3K、AKT、p-AKT、GRP78、CHOP和Caspase-12的蛋白表达水平.结果 与MCAO/R组比,中、高剂量TFR给药组大鼠神经行为学功能改善,脑梗死面积下降,脑水肿程度降低,脑皮质区病理损伤减轻,神经细胞凋亡率明显减少,抗凋亡蛋白Bcl-2表达升高,促凋亡蛋白Bax和cleaved Caspase-3表达降低,p-PI3K/PI3K、p-AKT/AKT表达升高.ERS相关蛋白GRP78、CHOP、Caspase-12表达降低.结论 TFR可通过调控PI3K/AKT信号通路和ERS途径抑制神经细胞凋亡,从而发挥对CIRI大鼠的保护作用.
Aim To observe the effect of corticotropin-releasing factor(CRF)-expressing neurons on presym-pathetic neurons in hypothalamic paraventricular nucle-us(PVN)of normotensive Wistar Kyoto(WKY)rats or spontaneously hypertensive rats(SHR),and to elu-cidate the underlying neuronal circuit mechanism of central sympathetic hyperexcitability.Methods The expression levels of CRF protein in WKY rats and SHR PVN were determined by Western blot.Meanwhile,the WKY and SHR PVN CRF-expressing neurons and presympathetic neurons were observed by immunofluo-rescent staining.Adult WKY rats and SHR were used in this study.By microinjection of Cre-dependent ade-no-associated viruses(AAV)that specifically recog-nized the CRF promoter and AAV of chemogenetics in-to the PVN,CRF-expressing neurons expressed desig-ner receptors exclusively activated by designer drugs(DREADDs).Human M3 muscarinic DREADD cou-pled to Gq receptor(hM3Dq)was specifically ex-pressed in PVN CRF-expressing neurons in WKY rats,while human M4 muscarinic DREADD coupled to Gi receptor(hM4Di)was specifically expressed in PVN CRF-expressing neurons in SHR.Clozapine-N-oxide(CNO),as a designer ligand,would couple to excita-tory hM3Dq or inhibitory hM4Di to regulate the excita-bility of PVN CRF-expressing neurons.Then the PVN presympathetic neurons were retrogradely labeled by microinjection of fluosecent tracer into the intermedio-lateral column(IML)of spinal cord.Lastly,whole cell patch clamp was used to determine the effect of CNO(10 μmol·L-1)on spontaneous excitatory postsynaptic currents(sEPSCs)and current-evoked firing of PVN presympathtic neurons of WKY rats and SHR.Results The expression of CRF protein in the PVN of SHR was significantly higher than that of WKY rats,and the activity and number of CRF-expressing neurons in the PVN of SHR were increased.PVN CRF-expressing neurons were expressed with chemoge-netic DREADDs and PVN presympathetic neurons were retrogradely labeled with fluorescent tracer in WKY rats and SHR.In SHR expressed with chemogenetic inhibi-tory hM4Di-mCherry of PVN CRF-expressing neurons,bath application of CNO to the brain slices resulted in a significant decrease in sEPSCs frequency,but no change in their amplitude of labeled PVN presympa-thetic neurons.In contrast,in WKY rats expressed with excitatory hM3Dq-eGFP of PVN CRF-expressing neurons,CNO had no obvious effect on the sEPSCs frequency and amplitude in PVN presympathetic neu-rons.Furthermore,bath application of CNO had no significant effect on current-evoked firing of PVN pres-ympathetic neurons of either WKY rats with hM3Dq-eGFP expression in CRF neurons or SHR with hM4Di-mCherry expression in CRF neurons.Conclusions The activity and number of PVN CRF-expressing neu-rons are increased in SHR,and CRF-expressing neu-rons enhance the excitability of presympathetic neu-rons,which acts as a regulatory neuronal microcircuit between CRF neurons and presympathetic neurons in the PVN.
Mother-to-child transmission of hepatitis B virus(HBV)is one of the main ways of transmission and the main cause of chronic hepatitis B after infection.Therefore,preven-ting mother-to-child transmission of HBV is particularly impor-tant in reducing the incidence of chronic hepatitis B.Currently,nucleoside(acid)analoids(Nas)used for mother-to-child blocking of HBV include lamivudine(LAM),tibivudine(LdT)and tenofovir fumarate(TDF).Propofol tenofovir fumarate(TAF)has also been used in pregnant chronic hepatitis B pa-tients.This paper summarizes the efficacy,safety and antiviral treatment indications and termination time of the above-men-tioned drugs in mother-to-child preventing to provide suggestions for the selection and rational application of mother-to-child pre-venting Nas.
Aim To study the effect of baicalin on the activation of NLRP3 inflammasomes in human fibro-blast like synoviocytes of rheumatoid arthritis(HFLS-RA)and its mechanism.Methods To confirm that baicalin alleviated the activation of NLRP3 inflamma-some in HFLS-RA,immunofluorescence was used to observe the expression of NLRP3 before and after ba-icalin treatment.Western blot was used to detect the protein expression of p-PI3K,p-Akt,NF-κB p65,NL-RP3,ASC and caspase-1 after baicalin treatment for 48 h,and ELISA was employed to detect the contents of IL-1 and IL-18 in the supernatents.In order to ex-plore the mechanism of baicalin alleviating the activa-tion of NLRP3 inflammasome,double luciferin and Westen blot analysis were applied to verify the corre-sponding relationship between let-7i-3p and PIK3CA.RT-qPCR was utilized to determine the expression of let-7i-3p and PI3K before and after baicalin interven-tion.let-7i-3p interference was used to verify whether baicalin mitigated the activation of enhanced NLRP3 inflammasomes.Results Baicalin(50,100 mg· L-1)significantly reduced the activation of NLRP3 in-flammasomes,inhibited the protein expressions of p-PI3K,p-Akt,NF-κB p65,NLRP3,ASC and caspase-1,and the secretion of IL-1 and IL-18.let-7i-3p and PIK3CA had a targeted correspondence,and baicalin up-regulated the expression of let-7i-3p and down-reg-ulated the expression of PIK3CA.Baicalin attenuated the activation of NLRP3 inflammasomes enhanced by let-7i-3p interference.Conclusions Baicalin can up-regulate let-7i-3p expression,inhibit PI3K/Akt/NF-κB signal transduction,and thus reduce the activation of NLRP3 inflammasomes in HFLS-RA.
