1.The Mechanisms of Quercetin in Improving Alzheimer’s Disease
Yu-Meng ZHANG ; Yu-Shan TIAN ; Jie LI ; Wen-Jun MU ; Chang-Feng YIN ; Huan CHEN ; Hong-Wei HOU
Progress in Biochemistry and Biophysics 2025;52(2):334-347
Alzheimer’s disease (AD) is a prevalent neurodegenerative condition characterized by progressive cognitive decline and memory loss. As the incidence of AD continues to rise annually, researchers have shown keen interest in the active components found in natural plants and their neuroprotective effects against AD. Quercetin, a flavonol widely present in fruits and vegetables, has multiple biological effects including anticancer, anti-inflammatory, and antioxidant. Oxidative stress plays a central role in the pathogenesis of AD, and the antioxidant properties of quercetin are essential for its neuroprotective function. Quercetin can modulate multiple signaling pathways related to AD, such as Nrf2-ARE, JNK, p38 MAPK, PON2, PI3K/Akt, and PKC, all of which are closely related to oxidative stress. Furthermore, quercetin is capable of inhibiting the aggregation of β‑amyloid protein (Aβ) and the phosphorylation of tau protein, as well as the activity of β‑secretase 1 and acetylcholinesterase, thus slowing down the progression of the disease.The review also provides insights into the pharmacokinetic properties of quercetin, including its absorption, metabolism, and excretion, as well as its bioavailability challenges and clinical applications. To improve the bioavailability and enhance the targeting of quercetin, the potential of quercetin nanomedicine delivery systems in the treatment of AD is also discussed. In summary, the multifaceted mechanisms of quercetin against AD provide a new perspective for drug development. However, translating these findings into clinical practice requires overcoming current limitations and ongoing research. In this way, its therapeutic potential in the treatment of AD can be fully utilized.
2.Effect of Yinchenhao Tang Combined with Yinchen Zhufu Tang on Treg/Th17 Cells in Vitro from Patients with Hepatitis B Virus-related Acute-on-chronic Liver Failure
Menghui ZENG ; Shan DU ; Nianhua TAN ; Jie PENG ; Bin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):43-51
ObjectiveTo study the regulatory effects of Yinchenhao Tang combined with Yinchen Zhufu Tang on the expression of regulatory T (Treg)/helper T 17 (Th17) cells cultured in vitro from the patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF). MethodsFresh peripheral blood was collected from the patients with HBV-ACLF for the separation of peripheral blood mononuclear cells (PBMCs). Immunomagnetic beads were used to isolate primary Treg and naive CD4+ T cells. After in vitro expansion, naive CD4+ T cells were induced to differentiate into Th17 cells. Rats were treated with the clearing method (Yinchenhao Tang), warming method (Yinchen Zhufu Tang), and combination of clearing method with warming method (Yinchenhao Tang combined with Yinchen Zhufu Tang, also known as Wenyang Jiedu Huayu Prescription), respectively, and then the medicated plasma samples were collected. Meanwhile, blank plasma was collected from the rats treated with normal saline. Cells were classified into blank, clearing method (5.04 g·kg-1), warming method (6.21 g·kg-1), and combination of clearing method with warming method (17.1 g·kg-1) groups and treated with corresponding plasma. The frequency of Treg/Th17 cells was detected by flow cytometry. The level of transforming growth factor-β (TGF-β) was measured by the enzyme-linked immunosorbent assay. The cytometric bead array (CBA) was employed to measure the levels of interleukin-10 (IL-10), interleukin-17A (IL-17A), tumor necrosis factor-α (TNF-α), and interleukin-23 (IL-23). The mRNA and protein levels of Forkhead box P3 (FoxP3) and retinoic acid-related orphan receptor-gamma t (ROR-γt) were determined by Real-time PCR and Western blot, respectively. ResultsCompared with the blank group, the combination of clearing method with warming method group showed decreased frequency of Treg and Th17 cells, lowered levels of Treg cytokines (TGF-β and IL-10) and Th17 cytokines (TNF-α, IL-17A, and IL-23), and down-regulated mRNA and protein levels of FoxP3 and ROR-γt (P<0.01). Compared with the clearing method group, the combination of clearing method with warming method group showed decreased Treg cell frequency and down-regulated mRNA and protein levels of FoxP3. Meanwhile, the combination group showed decreased Th17 cell frequency, lowered levels of TGF-β, IL-10, TNF-α, IL-17A, and IL-23, and down-regulated mRNA and protein levels of ROR-γt (P<0.05, P<0.01). Compared with the warming method group, the combination of clearing method with warming method group showed decreased frequency of Treg cells and down-regulated mRNA and protein levels of FoxP3. Meanwhile, the combination group showed decreased Th17 cell frequency, declined levels of TGF-β, IL-10, TNF-α, IL-17A, and IL-23, and down-regulated mRNA and protein levels of ROR-γt (P<0.05). ConclusionThe combination of clearing method with warming method can down-regulate the expression of specific cytokines of Treg and Th17 cells, inhibit the over activation of Treg and Th17 cells, and reduce the secretion of cytokines such as TGF-β, IL-10, TNF-α, IL-17A, and IL-23, thereby alleviating inflammation and improving the prognosis of the patients with liver failure.
3.Differences in Expression of Intestinal Flora in Patients with Different TCM Syndrome Types of "Yang Huang-Yin-Yang Huang-Yin Huang" in Hepatitis B Virus-related Acute-on-chronic Liver Failure
Shan DU ; Menghui ZENG ; Nianhua TAN ; Jie PENG ; Bin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):62-69
ObjectiveTo investigate the differential expression of intestinal flora in patients with different traditional Chinese medicine (TCM) syndrome types (Yang Huang syndrome, Yin-Yang Huang syndrome, and Yin Huang syndrome) of hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) and clarify the biological basis of jaundice and Yin Huang syndrome in liver failure. MethodsA total of 20 cases of HBV-ACLF patients were included in the Yang Huang group, 20 cases in the Yin-Yang Huang group, 16 cases in the Yin Huang group, and 20 healthy adult volunteers. 16S rRNA gene sequencing was used to detect the diversity, species distribution, and differences of the subjects' intestinal flora, and bioinformatics analysis was conducted. ResultsCompared with those in the healthy control group, the species richness and diversity of intestinal flora in the HBV-ACLF Yang Huang group, Yin-Yang Huang group, and Yin Huang group were significantly reduced, and there were significant differences in the composition of intestinal flora compared with healthy volunteers. However, there were no significant differences in the species richness, diversity, and composition of intestinal flora among the three groups. LEfSe analysis showed that compared with the healthy control group, the HBV-ACLF Yang Huang group showed significant enrichment of Staphylococcus aureus(P<0.01). Yin-Yang Huang group showed significant enrichment of s_Ileibacterium valens(P<0.05,P<0.01), and the Yin Huang group showed significant enrichment of Enterococcus faecium and Streptococcus sali varius(P<0.05). These strains may be biomarkers between the three groups of patients and the healthy control group. Compared with that in the Yin-Yang Huang group, Tyzzerella_nexilis was significantly enriched in the Yang-Huang group, and Streptococcus lactiae was significantly enriched in the Yin-Yang Huang group. Compared with that in the Yang-Huang group and the Yin-Yang Huang group, Enterococcus faecalis was significantly enriched in the Yin Huang group. The above strains may be biomarkers among the three groups of patients, and Enterococcus faecium may be a biomarker for the transition from the Yang Huang group to the Yin Huang group. ConclusionsThere are significant differences in the intestinal flora between patients with HBV-ACLF Yang Huang syndrome, Yin-Yang Huang syndrome, and Yin Huang syndrome. Enterococcus faecium is significantly enriched in the Yin Huang syndrome group, suggesting that dysbiosis of the intestinal flora may be the biological basis for jaundice and Yin Huang syndrome in liver failure.
4.Changes of Treg/Th17 Cell Expression in Peripheral Blood of Patients with Hepatitis B Virus-related Acute-on-chronic Liver Failure in Five Types of Traditional Chinese Medicine Syndrome
Shan DU ; Menghui ZENG ; Nianhua TAN ; Jie PENG ; Bin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):70-76
ObjectiveTo study the expression differences of regulatory T cells (Treg) and T helper 17 cells (Th17) in the peripheral blood of patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) in five types of traditional Chinese medicine (TCM) syndrome. MethodsA total of 144 patients with HBV-ACLF were included and divided into five types of TCM syndrome, including 34 cases of heat-toxin amassment syndrome, 44 cases of dampness-heat amassment syndrome, 27 cases of Qi-deficiency and stasis jaundice syndrome, 21 cases of spleen-kidney Yang deficiency syndrome, and 18 cases of liver-kidney Yin deficiency syndrome. Meanwhile, 30 healthy volunteers were included as controls. The frequency of Treg and Th17 cells in the peripheral blood of subjects in each group was detected by flow cytometry, and the Treg/Th17 ratio was calculated. Cytometric bead array (CBA) was used to detect the levels of cytokines interleukin (IL)-10, transforming growth factor-β (TGF-β), tumor necrosis factor-α (TNF-α), IL-17A, and IL-23. Real-time fluorescence quantitative PCR (Real-time PCR) detected the mRNA expression of forkhead box P3 (FoxP3) and retinoic acid-related orphan receptor gamma t (ROR-γt). Results(1) Compared with that in the healthy control group, the frequency of Treg and Th17 cells in patients with various TCM syndrome types of HBV-ACLF increased (P<0.05). Compared with that in the heat-toxin amassment syndrome group, the frequency of Treg and Th17 cells decreased in the dampness-heat amassment syndrome group (P<0.05), while the frequency of Treg and Th17 cells increased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the dampness-heat amassment syndrome group, the frequency of Treg and Th17 cells increased in the dampness-heat amassment syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the Qi-deficiency and stasis jaundice syndrome group, the frequency of Treg and Th17 cells increased in the spleen-kidney Yang deficiency syndrome group (P<0.05), while the frequency of Treg and Th17 cells decreased in the liver-kidney Yin deficiency syndrome group (P<0.05). Compared with the spleen-kidney Yang deficiency syndrome group, the frequency of Treg and Th17 cells decreased in the liver-kidney Yin deficiency syndrome group (P<0.05). (2) Compared with that in the healthy control group, the Treg/Th17 cell ratio in patients with various TCM syndromes of HBV-ACLF decreased (P<0.05). Compared with that in the heat-toxin amassment syndrome group, the Treg/Th17 cell ratio increased in the dampness-heat amassment syndrome group (P<0.05), while it decreased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the dampness-heat amassment syndrome group, the Treg/Th17 cell ratio decreased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the Qi-deficiency and stasis jaundice syndrome group, the Treg/Th17 cell ratio increased in the spleen-kidney Yang deficiency syndrome group and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with the spleen-kidney Yang deficiency syndrome group, the Treg/Th17 cell ratio in the liver-kidney Yin deficiency syndrome group increased (P<0.05). (3) Compared with those in the healthy control group, the levels of Treg-related cytokines IL-10 and TGF-β, as well as Th17-related cytokines TNF-α, IL-17A, and IL-23, were elevated in patients with various TCM syndrome types of HBV-ACLF (P<0.05). There was no significant difference in TNF-α levels among different TCM syndrome types. Compared with those in the heat-toxin amassment syndrome group, the levels of IL-10, TNF-β, IL-17A, and IL-23 in the dampness-heat amassment syndrome group, Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome groups increased (P<0.05). Compared with those in the dampness-heat amassment syndrome group, the levels of IL-10, TGF-β, IL-17A, and IL-23 increased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with those in the Qi-deficiency and stasis jaundice syndrome group, the levels of IL-10, TGF-β, IL-17A, and IL-23 in the spleen-kidney Yang deficiency syndrome group increased (P<0.05), while those in the liver-kidney Yin deficiency syndrome group decreased (P<0.05). Compared with those in the spleen-kidney Yang deficiency syndrome, the levels of IL-10, TGF-β, IL-17A, and IL-23 in the liver-kidney Yin deficiency syndrome group decreased (P<0.05). (4) Compared with that in the healthy control group, the mRNA of Treg/Th17 cell specific transcription factors FoxP3 and ROR-γt were elevated in patients with various TCM syndrome types of HBV-ACLF (P<0.05). Compared with that in the heat-toxin amassment syndrome group, the mRNA of FoxP3 and ROR-γt increased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the dampness-heat amassment syndrome group, the mRNA of FoxP3 and ROR-γt increased in the Qi-deficiency and stasis jaundice syndrome group, spleen-kidney Yang deficiency syndrome group, and liver-kidney Yin deficiency syndrome (P<0.05). Compared with that in the Qi-deficiency and stasis jaundice syndrome group, the mRNA of FoxP3 and ROR-γt in the spleen-kidney Yang deficiency syndrome group increased (P<0.05), and it decreased in the liver-kidney Yin deficiency syndrome group (P<0.05). Compared with that in the spleen-kidney Yang deficiency syndrome group, the mRNA of FoxP3 and ROR-γt decreased in the liver-kidney Yin deficiency syndrome group (P<0.05). ConclusionThe frequency and ratio of Treg/Th17 cells, as well as the expression of related cytokines and specific receptors in peripheral blood of patients with HBV-ACLF in five types of TCM syndromes are different, which has certain reference value for TCM syndrome differentiation and treatment of patients with HBV-ACLF.
5.Early outcomes of anterior segment parameters in patients with high myopia after implantable collamer lens V4c implantation
Bingxin PAN ; Jie WU ; Pengfei JIANG ; Shan XU ; Kun HE
International Eye Science 2024;24(3):491-494
AIM: To study the early outcomes of anterior segment parameters after implantation of an implantable collamer lens with a central hole(ICL V4c)in patients with high myopia.METHODS:A total of 82 cases(160 eyes)with high myopia, including 42 males(82 eyes)and 40 females(78 eyes), aged 26.0±4.6(21 to 37)years, who underwent ICL V4c implantation at our institution from February 2019 to September 2022 and were followed up for 1 a, were included. The general characteristics of the anterior segment of the eye were measured preoperatively: spherical equivalent, mean horizontal corneal curvature, white-to-white(WTW), and axial length(AL); intraocular pressure(IOP), endothelial cell density(ECD), central anterior chamber depth(CACD), anterior chamber volume(ACV)and anterior chamber angle(ACA)were measured preoperatively and at 1 d, 1 wk, 1, 3 and 6 mo postoperatively. Furthermore, the distance from the centre of the posterior surface of the ICL V4c optical zone to the anterior surface of the lens(vault)was measured at 1 d, 1 wk, 1, 6 mo, and 1 a after surgery.RESULTS: The mean preoperative spherical equivalent of the patients was -7.56±2.55 D, mean horizontal corneal curvature was 42.89±1.47 D, WTW was 11.64±0.37 mm, and AL was 26.64±0.93 mm. The baseline IOP was 15.97±2.13 mmHg, and the differences in IOP at each time point after ICL V4c implantation compared to preoperative were not statistically significant(F=0.875, P=0.504); ECD was 2 989.30±140.78 cells/mm2 at baseline, and ECD at 6 mo after ICL V4c implantation was not statistically significant compared with preoperative ECD(t=1.475, P=0.142); CACD was 3.19±0.21 mm at baseline, and ACV was 210.30±27.7 mm3, and CACD and ACV were significantly lower than preoperative at all postoperative time points(F=111.10, 288.38, all P<0.001). The baseline ACA was 35.44°±11.27°, and the ACA at each time point after ICL V4c implantation was significantly lower than preoperatively(F=21.23, P<0.001). The vault was 665.32±184.03 μm at 1 d postoperatively, and continued to be significantly reduced at 1 wk, 1, 6 mo, and 1 a postoperatively compared with 1 d(F=52.10, P<0.001). However, it remained stable at 6 mo and 1 a postoperatively, and the difference was not statistically significant compared with vault at 1 mo postoperatively(P>0.05).CONCLUSION: ICL V4c has certain safety and efficiency in 1 a postoperative follow-up, and the parameters of the anterior segment of the eye stabilized in the early period.
6.Use of Thread-hanging Microwave Antenna to Assist in Precise Puncture of Lung Nodule and Influencing Factors
Hui JIN ; Jie TAN ; Shan SONG ; Yinghu WANG ; Lihua MA
Cancer Research on Prevention and Treatment 2024;51(2):110-114
Objective To explore a precise method with a microwave antenna for puncture of pulmonary nodules and analyze phenomena that affect the puncture results. Methods Clinical data of 107 cases with solitary malignant pulmonary nodules were collected, and the mean length of pulmonary nodules was 13.6±0.6 mm in CT axial position. A thread-hanging method was used to assist the puncture of pulmonary nodules. The procedure was successful when the needle was not withdrawn and inserted into the central region of the nodule. The success rate and complications of the pulmonary procedure were recorded. The incidence of the following phenomena were also documented: needle coercing, needle slipping, needle tip pushing, pulmonary nodule prolapsing, radial nodule deformation, nodular masking, and radial movement distance of needle tip. Results In all of 107 cases evaluated, the antenna puncture was successful in 101 cases (94.4%) but failed in 6 cases (5.6%). Pneumothorax and pulmonary hemorrhage occurred in 23 (21.5%) and 19 cases (17.8%), respectively. The following phenomena occurred: needle coercing in 9 cases (8.4%), needle slipping in 6 cases (5.6%), needle tip pushing in 19 cases (17.8%), pulmonary nodule prolapsing in 15 cases (14%), radial nodule deformation in 14 cases (13.1%), and nodular masking in 5 cases (4.7%). The mean radial adjusting distance of needle tip was 0.7±0.4 cm. Conclusion The thread-hanging method can assist in the accurate puncture of microwave antenna for pulmonary nodules. We should focus and deal with phenomena that may occur and affect the result of puncture.
7.Inhibitory effects of Ginkgo biloba extract on renal inflammation in diabetic nephropathy model mice and its mechanism
Jing CHEN ; Xiaoyi YANG ; Jing CHEN ; Xin SHAN ; Jie WANG ; Huiqin XU ; Zhiyang LYU
China Pharmacy 2024;35(2):186-191
OBJECTIVE To investigate the inhibitory effects of Ginkgo biloba extract (GBE) on renal inflammation in diabetic nephropathy (DN) model mice, and its potential mechanism. METHODS KK/Ay mice were fed with high fat and high sugar to induce DN model. They were divided into model group, positive control group [metformin 200 mg/(kg·d)], GBE low-dose and high-dose groups [100, 200 mg/(kg·d)], with 6 mice in each group. Six C57BL/6J mice were fed with a regular diet as the control group. Administration groups were given relevant liquid intragastrically, control group and model group were given constant volume of normal saline intragastrically, once a day, for 8 consecutive weeks. The body weight, fasting blood glucose, 24-hour food intake, 24-hour urine output, monocyte chemoattractant protein-1 (MCP-1), interleukin-12 (IL-12), IL-10, advanced glycation end products (AGEs), blood urea nitrogen (BUN) and serum creatinine (Scr) of mice were measured, and the ratio of bilateral kidneys to body weight was also calculated. The pathological injury and fibrotic changes of the renal cortex were observed, and the expressions of macrophage polarization marker proteins [type M1: inducible nitric oxide synthase (iNOS); type M2: arginase-1 (Arg-1)] and AGEs-the receptor of advanced glycation end products (RAGE)/Ras homolog gene pharm_chenjing@163.com family member A (RhoA)/Rho-associated coiled-coil forming protein kinase (ROCK) signaling pathway-related proteins were determined in renal cortex. RESULTS Compared with the model group, the symptoms such as renal cortical hyperplasia, vacuoles, infiltration of inflammatory cells, and renal cortical fibrosis had been improved in GBE low-dose and high-dose groups; body weight, serum level of IL-10, the expression of Arg-1 in the renal cortex were significantly higher than model group (P< 0.01); fasting blood glucose, 24-hour food intake, 24-hour urine output, serum levels of MCP-1, IL-12, BUN, Scr and AGEs, the ratio of bilateral kidneys to body weight, renal injury score, the proportion of renal interstitial fibrosis, the protein expressions of iNOS, RAGE, RhoA and ROCK1 (except for GBE low-dose group) in renal cortex were significantly lower than model group (P<0.01). CONCLUSIONS GBE could improve kidney damage and alleviate inflammatory response in DN model mice, the mechanism of which may be related to inhibiting the AGEs-RAGE/RhoA/ROCK signaling pathway and regulating macrophage polarization.
8.Clinical Observation on Abdominal Penetrating Moxibustion Combined with Acupuncture at Four Chong Acupoints for the Treatment of Stroke in the Convalescent Stage
Nan YU ; Jie HU ; Hai-Ning LI ; Shan-Shan SONG ; De-Song ZHENG ; Xin-Hong XUE ; Ying GAO
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(1):116-122
Objective To observe the effects of abdominal penetrating moxibustion combined with acupuncture at the"four chong points"on balance,walking function and trunk control in patients recovering from stroke.Methods Seventy-eight patients recovering from stroke were randomly divided into an observation group and a control group,with 39 patients in each group.The control group was given conventional rehabilitation exercises,while the observation group was given abdominal penetrating moxibustion combined with acupuncture at the"four chong points"on the basis of the control group.Both groups were treated for 2 consecutive months.After 2 months of treatment,the clinical efficacy of the two groups was evaluated,and the changes in the Berg Scale score and the Timed Up and Go Test(TUGT)were observed before and after treatment.The changes in the National Institutes of Health Stroke Scale(NIHSS)scores were compared before and after treatment between the two groups.The Sheikh Trunk Control Scale scores were also evaluated.Results(1)The total effective rate of the observation group was 94.87%(37/39),and the total effective rate of the control group was 80.00%(31/39),and the efficacy of the observation group was superior to that of the control group,and the difference was statistically significant(P<0.05).(2)After treatment,the Berg scores of the patients in the two groups were significantly increased(P<0.05),and the Berg scores of the observation group were higher than those of the control group,and the difference was statistically significant(P<0.05).(3)After treatment,the TUGT time and NIHSS score of patients in the two groups were significantly improved(P<0.05),and the TUGT time of the observation group was shorter than that of the control group,and the NIHSS score was lower than that of the control group,and the difference was statistically significant(P<0.05).(4)After treatment,the Sheikh trunk control scores of the two groups were significantly increased(P<0.05),and the Sheikh trunk control score of the observation group was higher than that of the control group,and the difference was statistically significant(P<0.05).Conclusion Abdominal penetrating moxibustion method combined with acupuncture at the four chong points for the treatment of stroke recovery can effectively restore the patients'balance and walking function,improve the patients'trunk control ability,and the therapeutic effect is precise.
9.Moxibustion inhibits NLRP3/Caspase-1 pathway mediated cell pyroptosis and alleviates cerebral ischemia/reperfusion injury
Yanyan YU ; Juan LIU ; Yue YANG ; Qianhui WANG ; Shan LI ; Jie JIANG
Chinese Journal of Tissue Engineering Research 2024;28(34):5473-5479
BACKGROUND:It was found that moxibustion can inhibit the inflammatory factors in the serum of rats with cerebral ischemia/reperfusion injury,resist oxidative stress,inhibit cell apoptosis,and effectively reduce cerebral ischemia/reperfusion injury. OBJECTIVE:To observe the effects of different moxibustion intervention time on the expression levels of nucleotide binding oligomerization domain-like protein 3 inflammasome(NLRP3),cysteine aspartase(caspase-1),apoptosis-related speck-like protein,exfoliatin-D protein,interleukin-1β and interleukin-18 in rats with cerebral ischemia/reperfusion injury,and to explore its action mechanism. METHODS:SD rats were randomly divided into sham operation group(n=9)and operation group(n=36).The model of focal cerebral ischemia/reperfusion injury was established by middle cerebral artery occlusion in the operation group.After successful modeling,the rats in the operation group were further divided into model group,moxibustion 10-minute group,moxibustion 15-minute group and moxibustion 30-minute group,with 9 rats in each group.Rats in the moxibustion 10-minute,15-minute and 30-minute groups were given moxibustion at"Baihui,Dazhui and Zusanli",respectively,once a day for a total of 7 days.The neurological deficits of rats were evaluated by LONGA method.The cerebral infarction was observed by 2,3,5-triphenyltetrazolium chloride staining.The pathological changes of brain tissue were observed by hematoxylin-eosin staining.The contents of interleukin-1β and interleukin-18 in serum of rats in each group were detected by ELISA.Immunohistochemistry and western blot assay were used to detect the expression levels of NLRP3,caspase-1,apoptosis-related spot-like protein and gasdermin D in the ischemic cortex of rats in each group. RESULTS AND CONCLUSION:Compared with the sham operation group,the neurological deficit score of the model group was significantly increased(P<0.01).Compared with the model group,the neurological deficit score of the moxibustion groups was significantly reduced(P<0.01).Compared with the sham operation group,the infarct volume of the model group was significantly increased(P<0.01).Compared with the model group,the infarct volume of the moxibustion groups was significantly reduced(P<0.01);the infarct volume of the rats was smallest in the moxibustion 30-minute group(P<0.05).Compared with the model group,the contents of inflammatory factors interleukin-1β and interleukin-18 in the serum of rats in the moxibustion groups were decreased(P<0.01).Compared with the moxibustion 10-minute group,the contents of inflammatory factors in the serum of rats in the moxibustion 30-minute group were significantly decreased(P<0.05).Compared with the model group,the expression of NLRP3,apoptosis-related spot-like protein,Caspase-1 and gasdermin D protein in the ischemic cortex of the moxibustion groups was significantly decreased(P<0.01).Compared with the moxibustion 10-minute and 15-minute groups,the expression of protein in the moxibustion 30-minute group was significantly decreased(P<0.05).It is concluded that moxibustion at Baihui,Dazhui and Zusanli can reduce cerebral ischemia/reperfusion injury,among which moxibustion for 30 minutes has the best effect,and its mechanism may be related to the inhibition of pyroptosis mediated by NLRP3/Caspase-1 pathway.
10.Hypericin inhibits the expression of NLRP3 in microglia of Parkinson's disease mice and alleviates the damage of DA-ergic neurons
Li-Shan FAN ; Jia ZHANG ; Si-Xiang NIU ; Qi XIAO ; Hui-Jie FAN ; Lei XU ; Li-Xia YANG ; Lu JIA ; Shao-Chen QIN ; Bao-Guo XIAO ; Cun-Gen MA ; Zhi CHAI
The Chinese Journal of Clinical Pharmacology 2024;40(17):2523-2527
Objective To observe the intervention effect of hypericin(HYP)on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced Parkinson's disease(PD)mice model and its mechanism.Methods Thirty C57BL/6 mice were randomly divided into normal,model and experimental groups with 10 mice per group.PD mouse model was established after 7 days of intraperitoneal injection of MPTP,and drug intervention was carried out from the first day of modeling.Normal group and model group were intraperitoneally injected with 500 μL·kg·d-1 0.9%NaCl.The experimental group was intraperitoneally injected with 25 mg·kg·d-1 HYP.The three groups of rats were given the drug once each time for 14 days.The expression levels of tyrosine hydroxylase(TH),Nod-like receptor thermal protein domain protein 3(NLRP3)and ionized calcium binding adapter molecule 1(Iba1)in the striatum of nigra were detected by Western blot.Results The climbing time of normal,model and experimental groups was(5.35±0.43),(9.71±1.19)and(8.07±0.34)s;suspension scores were(2.92±0.15),(1.38±0.28)and(1.96±0.28)points;the relative expression levels of TH protein were 1.04±0.06,0.51±0.09 and 0.75±0.07;the relative expression levels of NLRP3 protein were 0.51±0.03,1.00±0.04 and 0.77±0.06;the relative expression levels of Iba1 protein were 0.68±0.10,1.30±0.28 and 0.89±0.05,respectively.The above indexes in the model group were statistically significant compared with the experimental group and the normal group(all P<0.01).Conclusion HYP plays a therapeutic role in PD by inhibiting the expression of NLRP3 inflammasome in PD mice.

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