1.Mechanisms of Spermatogenic Dysfunction and Sperm Quality Degradation Induced by Propylthiouracil via Endoplasmic Reticulum Stress in Rats and Interventional Effect of Shugan Bushen Yulin Decoction
Yuhao MENG ; Xiaocui JIANG ; Min XIAO ; Chaoyang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):79-89
ObjectiveTo investigate the mechanisms through which Shugan Bushen Yulin decoction ameliorates spermatogenic dysfunction and sperm quality degradation caused by propylthiouracil (PTU) via the endoplasmic reticulum stress pathway in rats. MethodsSixty male rats were randomly assigned into six groups: a control group, a model group, low- (6.75 g·kg-1), medium- (13.5 g·kg-1), and high-dose (27 g·kg-1) Shugan Bushen Yulin decoction groups, and an L-carnitine (0.27 g·kg-1) group, with ten rats in each group. Other groups except the control group were administrated with PTU (10 mg·kg-1) by gavage for 12 consecutive days. After the completion of modeling, rats were administrated with corresponding agents or normal saline (control group) via gavage for 28 consecutive days. Twenty-four hours after the last administration, rats were sacrificed, and the body and organ (testis, epididymis, and seminal vesicle) weights were measured to calculate the organ indices. Hematoxylin-eosin staining was employed to observe the pathological changes in the testes and epididymis, and the testicular spermatogenic function of rats was scored. Enzyme-linked immunosorbent assay was employed to measure the levels of thyroid-stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4), follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), estradiol (E2), and malondialdehyde (MDA) and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the rat serum. Terminal deoxynucleotidyl transferase dUTP nick end labeling was adopted to assess the rate of testicular cell apoptosis. Western blot was conducted to determine the expression levels of glucose-regulated protein 78 (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), phosphorylated protein kinase RNA-like endoplasmic reticulum kinase (p-PERK), protein kinase RNA-like endoplasmic reticulum kinase (PERK), phosphorylated eukaryotic translation initiation factor 2α kinase (p-EIF2α), eukaryotic translation initiation factor 2α kinase (EIF2α), and activating transcription factor 4 (ATF4) in the testicular tissue of rats. ResultsCompared with the control group, the model group exhibited reductions in volumes and indexes of testes, epididymides, and seminal vesicles (P<0.01), pathological damage of testes and epididymides, and declines in spermatogenic function and sperm density and motility (P<0.01). In addition, the model group demonstrated elevated serum levels of TSH and MDA, lowered levels of T3, T4, FSH, LH, T, and E2, decreased activities of SOD and GSH-Px (P<0.05, P<0.01), an increased apoptosis rate of testicular cells (P<0.01), and up-regulated expression of GRP78, CHOP, p-PERK/PERK, p-EIF2α/EIF2α, and ATF4 in the testicular tissue (P<0.01). Compared with the model group, Shugan Bushen Yulin decoction at different doses improved the morphology and indexes of testes, epididymides, and seminal vesicles (P<0.05), restored the spermatogenic function (P<0.05, P<0.01), increased the sperm density and motility (P<0.05, P<0.01), and decreased the percentage of apoptotic testicular cells (P<0.05). Furthermore, the decoction lowered the serum level of MDA, elevated the serum levels of FSH, LH, T, and E2, and increased the activities of SOD and GSH-Px (P<0.05, P<0.01). Moreover, the expression levels of GRP78, CHOP, p-PERK/PERK, p-EIF2α/EIF2α, and ATF4 in the testicular tissue were down-regulated (P<0.05, P<0.01). ConclusionShugan Bushen Yulin decoction can ameliorate the spermatogenic dysfunction and sperm quality degradation induced by PTU in a rat model of hypothyroidism by repairing the testicular oxidative damage and regulating the PERK/EIF2α/ATF4 signaling pathway, thereby alleviating spermatogenic cell apoptosis.
2.Effect of Zuoguiwan on Development of Skin Barrier in Neonatal Rat Model of Congenital Kidney Deficiency Based on Intercellular Connections
He YU ; Min XIAO ; Xiaocui JIANG ; Min ZHAO ; Yinjuan LYU ; Jian GONG ; Jigang CAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):11-18
ObjectiveTo study the effect of Zuoguiwan on the development of skin barrier in the neonatal rat model of congenital kidney deficiency and unveil the underlying mechanism. MethodsSixty rats were paired in a female-to-male ratio of 2∶1, and the pregnant rats were assigned into control, congenital kidney deficiency, and low- and high-dose (2 and 8 g·kg-1, respectively) Zuoguiwan groups. The pregnant rats in other groups except the control group were exposed to chronic unpredictable mild stress for the modeling of congenital kidney deficiency. The rats in the control group and congenital kidney deficiency group were administrated with normal saline by gavage, and those in Zuoguiwan groups with Zuoguiwan suspension by gavage from day 1 of pregnancy. The serum level of interleukin-6 (IL-6) in the neonatal rats on the day of birth was determined by enzyme-linked immunosorbent assay. The full-thickness skin of neonatal rats on the day of birth was removed from the same position on the back and stained with hematoxylin-eosin for observation of histopathological changes, measurement of skin thickness, and counting of hair follicles. Terminal deoxynucleotidyl transferase-mediated nick end labeling was used to detect the apoptosis of skin tissue cells. The expression of interleukin-6 receptor (IL-6R) and interleukin-17A (IL-17A) in the skin tissue was assessed by immunohistochemistry and Western blot, and the expression of occludin, connexin 43 (Cx43), and zonula occludens-1 (ZO-1) in the skin tissue was assessed by immunofluorescence and Western blot. ResultsCompared with those in the control group, the neonatal rats in the congenital kidney deficiency group showed a rise in the serum IL-6 level (P<0.01), decreases in stratum corneum thickness, skin thickness, and number of hair follicles (P<0.01), increases in the expression of IL-6R and IL-17A in the skin tissue (P<0.01) and the number of apoptotic cells (P<0.01), and decreases in the expression of occludin, Cx43, ZO-1 (P<0.05). Compared with the congenital kidney deficiency group, the low- and high-dose Zuoguiwan groups showed declines in serum IL-6 level (P<0.05). The low-dose group showed increased number of hair follicles (P<0.05), and the high-dose group presented thickened stratum corneum (P<0.01), increased number of hair follicles (P<0.01), and down-regulated expression of IL-6R and IL-17A in the skin tissue (P<0.05, P<0.01). Both Zuoguiwan groups showcased decreased number of apoptotic cells (P<0.05, P<0.01), and the high-dose group showed up-regulated expression of occludin, Cx43, and ZO-1 in the skin tissue (P<0.05, P<0.01). ConclusionZuoguiwan can reduce the levels of IL-6 in the serum and IL-6R and IL-17A in the skin tissue and improve the expression of intercellular junction proteins, thereby ameliorating the abnormal development of the skin barrier in the neonatal rat model of congenital kidney deficiency.
3.Effect of Zuoguiwan on Development of Skin Barrier in Neonatal Rat Model of Congenital Kidney Deficiency Based on Intercellular Connections
He YU ; Min XIAO ; Xiaocui JIANG ; Min ZHAO ; Yinjuan LYU ; Jian GONG ; Jigang CAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):11-18
ObjectiveTo study the effect of Zuoguiwan on the development of skin barrier in the neonatal rat model of congenital kidney deficiency and unveil the underlying mechanism. MethodsSixty rats were paired in a female-to-male ratio of 2∶1, and the pregnant rats were assigned into control, congenital kidney deficiency, and low- and high-dose (2 and 8 g·kg-1, respectively) Zuoguiwan groups. The pregnant rats in other groups except the control group were exposed to chronic unpredictable mild stress for the modeling of congenital kidney deficiency. The rats in the control group and congenital kidney deficiency group were administrated with normal saline by gavage, and those in Zuoguiwan groups with Zuoguiwan suspension by gavage from day 1 of pregnancy. The serum level of interleukin-6 (IL-6) in the neonatal rats on the day of birth was determined by enzyme-linked immunosorbent assay. The full-thickness skin of neonatal rats on the day of birth was removed from the same position on the back and stained with hematoxylin-eosin for observation of histopathological changes, measurement of skin thickness, and counting of hair follicles. Terminal deoxynucleotidyl transferase-mediated nick end labeling was used to detect the apoptosis of skin tissue cells. The expression of interleukin-6 receptor (IL-6R) and interleukin-17A (IL-17A) in the skin tissue was assessed by immunohistochemistry and Western blot, and the expression of occludin, connexin 43 (Cx43), and zonula occludens-1 (ZO-1) in the skin tissue was assessed by immunofluorescence and Western blot. ResultsCompared with those in the control group, the neonatal rats in the congenital kidney deficiency group showed a rise in the serum IL-6 level (P<0.01), decreases in stratum corneum thickness, skin thickness, and number of hair follicles (P<0.01), increases in the expression of IL-6R and IL-17A in the skin tissue (P<0.01) and the number of apoptotic cells (P<0.01), and decreases in the expression of occludin, Cx43, ZO-1 (P<0.05). Compared with the congenital kidney deficiency group, the low- and high-dose Zuoguiwan groups showed declines in serum IL-6 level (P<0.05). The low-dose group showed increased number of hair follicles (P<0.05), and the high-dose group presented thickened stratum corneum (P<0.01), increased number of hair follicles (P<0.01), and down-regulated expression of IL-6R and IL-17A in the skin tissue (P<0.05, P<0.01). Both Zuoguiwan groups showcased decreased number of apoptotic cells (P<0.05, P<0.01), and the high-dose group showed up-regulated expression of occludin, Cx43, and ZO-1 in the skin tissue (P<0.05, P<0.01). ConclusionZuoguiwan can reduce the levels of IL-6 in the serum and IL-6R and IL-17A in the skin tissue and improve the expression of intercellular junction proteins, thereby ameliorating the abnormal development of the skin barrier in the neonatal rat model of congenital kidney deficiency.
4.Effect and Mechanism of Action of Epimedii Folium Polysaccharides on Mice with Exercise-induced Fatigue Based on p38 MAPK/NF-κB Signaling Pathway
Yue ZOU ; Min XIAO ; Yuhao MENG ; Kunyang TANG ; Xiaocui JIANG ; Zhipeng FANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):20-28
ObjectiveTo study the effects of Epimedii Folium polysaccharides on mice with exercise-induced fatigue and explore its possible mechanism of action. MethodICR male mice screened by swimming training were randomly divided into a control group, model group, vitamin C group, and low, medium, and high dose groups of Epimedii Folium polysaccharides, with eight mice in each group. The exercise-induced fatigue model was established by weight-bearing swimming training in each group except for the control group. After two weeks of weight-bearing swimming, the Epimedii Folium polysaccharide groups were given 100, 200, 400 mg∙kg-1 of Epimedii Folium polysaccharides by gavage, and the vitamin C group was given 200 mg∙kg-1 of vitamin C by gavage. The control group and the model group were given equal amounts of saline for 14 d. At the end of the experimental period, the body mass of the mice in each group and the time of last swimming due to exhaustion were recorded. Serum urea nitrogen (BUN), lactic acid (LA), lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidation (GSH-Px), myoglycogen (MG) in skeletal muscle, hepatic glycogen (HG) in the liver were detected by kits. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in muscle tissue. Western blot was used to detect the protein expression of p38 mitogen-activated protein kinase (p38 MAPK), phosphorylation (p)-p38 MAPK, extracellular signal-regulated kinase1/2 (ERK1/2), nuclear factor-κB (NF-κB), p-NF-κB, interleukin-1β (IL-1β), and interleukin-6 (IL-6) in muscle tissue. The immunofluorescence (IF) method was used to detect the expression of tumor necrosis factor-α (TNF-α) in skeletal muscle tissue of mice in each group. ResultCompared with the control group, the body mass of mice in the model group decreased, and the time of last swimming due to exhaustion decreased (P<0.01). In addition, there were significantly higher serum levels of the fatigue metabolites LA, LDH, BUN, and lipid peroxidation product MDA (P<0.01) and decreased levels of MG, HG, SOD, and GSH-Px (P<0.01). The protein expressions of p-p38 MAPK, ERK1/2, p-NF-κB, IL-1β, IL-6, and TNF-α in skeletal muscle tissue were significantly higher than those of the control group (P<0.01). Compared with the model group, the body mass and time of last swimming due to exhaustion of the mice in the low, medium, and high dose groups of Epimedii Folium polysaccharides and the vitamin C group were increased (P<0.05, P<0.01), and the contents of LA, LDH, BUN, and MDA were significantly decreased (P<0.05, P<0.01). The levels of MG, HG, SOD, and GSH-Px increased (P<0.05, P<0.01), and the protein expression levels of p-p38 MAPK, ERK, p-NF-κB, IL-1β, IL-6, and TNF-α in skeletal muscle tissue decreased (P<0.05, P<0.01). ConclusionEpimedii Folium polysaccharides can play a role in alleviating exercise-induced fatigue by inhibiting the p38 MARK/NF-κB signaling pathway, thereby reducing the accumulation of metabolites, improving the activity of antioxidant enzymes, increasing the glycogen content of the body, and reducing inflammation in skeletal muscle.
5.Regulatory Mechanism of Drug-Containing Serum of Jinghou Zengzhi Prescription on GDF9 Expression and Apoptosis of Ovarian Granulosa Cells in Rats with Controlled Ovarian Hyperstimulation
Zhen YANG ; Xiao-Yan CHEN ; Shao-Ru JIANG ; Shu-Zhu YE ; Xiao-Hong FANG ; Wei-Min DENG ; Xin-Yu GUO
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(3):735-741
Objective To observe the regulatory mechanism of drug-containing serum of Jinghou Zengzhi Prescription based on qi and blood replenishing method on the expression of growth and differentiation factor 9(GDF9)and apoptosis of ovarian granulosa cells in rats with controlled ovarian hyperstimulation(COH).Methods Serum of COH rats(blank serum)and serum of COH rats gavaged by the Jinghou Zengzhi Prescription were prepared.A COH rat model was established and ovarian granulosa cells were collected.The experiment was divided into 5 groups:blank serum group,drug-containing serum group,drug-containing serum+SB203580[p38 mitogen-activated protein kinase(p38MAPK)inhibitor]group,drug-containing serum + PDTC[nuclear transcription factor κB(NF-κB)inhibitor]group,drug-containing serum + SB203580 + PDTC group.The mRNA expression levels of p38MAPK,casein kinase 2(CK2),nuclear transcription factor κB inhibitor α(IκBα),NF-κB and GDF9 were detected by real-time quantitative polymerase chain reaction(qRT-PCR),and GDF9 protein expression level was detected by Western Blot,and ovarian granulosa cell apoptosis was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL).Results The drug-containing serum of Jinghou Zengzhi Prescription decreased the mRNA expressions of p38MAPK and NF-κB,elevated the mRNA expressions of CK2 and IκBα,increased the mRNA and protein expression levels of GDF9,and decreased the apoptosis rate of ovarian granulosa cells in COH rats.The addition of p38MAPK inhibitor SB203580 alone and the addition of NF-κB inhibitor PDTC alone both promoted the mRNA and protein expressions of GDF9 and reduced the apoptosis rate of granulosa cells.Conclusion The drug-containing serum of Jinghou Zengzhi Prescription based on qi and blood replenishing method can promote the expression of GDF9 and inhibit the apoptosis of ovarian granulosa cells in rats with COH,and its mechanism may be related to the regulation of the expression of genes of the dual signaling pathways of p38MAPK and NF-κB.
6.Analysis on Nutritional Status of Vitamin D Among Adults of 6 Ethnic Minority Groups Native to Yunnan Province
Nan ZHANG ; Yanling YANG ; Qingqing WAN ; Jiang ZHAO ; Min PENG ; Xiao ZHU ; Zhitao LIU ; Qiang ZHANG
Journal of Kunming Medical University 2024;45(2):105-111
Objective To assess the nutritional status of vitamin D and associated factors among adults of six ethnic minority groups native to Yunnan Province,and provide evidence for policy making.Methods Between May 2019 and August 2020,a total of 690 adults were selected from Jinuo,Bulang,Jingpo,Deang,Achang and Pumi ethnic groups according to the sex and age composition in the 6th national census.A questionnaire survey and an anthropometric examination were conducted by trained health workers,and serum 25(OH)D levels were determined with high-performance liquid chromatography-tandem mass spectrometry.Results The median of serum 25(OH)D was 28.7(P25~P75∶24.3~33.8)ng/mL,and the prevalence of vitamin D sufficiency,insufficiency and deficiency were 44.2%、47.5%and 8.3%,respectively.There were significant differences in serum 25(OH)D levels among the six ethnic groups(χ2=139.29,P<0.01).Multivariate logistic regression showed that ethnic groups living in higher latitude areas(Pumi,Jingpo,Deang,and Achang),women,and those whose BMI≥24.0 were more likely to be vitamin D insufficient or deficient.Conclusion More than half of the ethnic adults suffer from vitamin D malnutrition which also varies across ethnicities.Further surveillance and interventions among key areas and populations are needed.
7.Zuoguiwan Prevent Decreased Ovarian Reserve in Prenatal Stressed Offspring Rats via TLR4/NF-κB p65 Signaling Pathway
Yinjuan LYU ; Weiheng ZHANG ; Siyi CHEN ; Wenjing TIAN ; Jian GONG ; Yuhao MENG ; Huan HE ; Xiaocui JIANG ; Min ZHAO ; Min XIAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(17):67-76
ObjectiveTo observe the effect of Zuoguiwan on ovarian reserve in the female offspring rat model of prenatal stress (PS) and explore the mechanism based on Toll-like receptor 4/nuclear factor-κB p65 (TLR4/NF-κB p65) signaling pathway. MethodThirty-two pregnant rats were prepared and randomized into four groups (n=8): control, model, Zuoguiwan (18.9 mg·kg-1), and vitamin E (1.44 mg·kg-1). Except the control group, the other three groups were subjected to chronic unpredictable mild stress (CUMS) from day 11 of pregnancy, and the modeling was accompanied by gavage with corresponding drugs until delivery. The PS model was evaluated by the sucrose preference test, open field test, and serum corticosterone (CORT) level. The estrous cycle was monitored and the morphological changes in the ovarian tissue were observed. The serum levels of estradiol (E2), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and anti-Mullerian hormone (AMH) in the 75-day-old offspring rats were measured by enzyme-linked immunosorbent assay (ELISA) to evaluate the ovarian reserve. The ovary and uterus indices were calculated. The serum levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). The morphology of the ovarian tissue in the offspring on the day of birth and day 75 after birth was observed by hematoxylin-eosin staining. The transport of NF-κB p65 to the nucleus in the ovaries of the 75-day-old offspring was detected by the immunofluorescence (IF) assay. The expression of TLR4, NF-κB p65 and other related proteins in the ovarian tissue was determined by Western blot. ResultCompared with the control group, the model group showed reduced primordial follicles in the offspring on the day of birth (P<0.01) as well as disturbed estrous cycle, decreased ovary index and uterus index (P<0.01), reduced corpus luteum, increased atretic follicles (P<0.01), lowered serum levels of AMH and E2 (P<0.01), elevated serum levels of LH, FSH, IL-1β, and TNF-α (P<0.05, P<0.01), and up-regulated protein levels of TLR4, NF-κB p65, recombinant myeloid differentiation factor 88 (MyD88), and phosphorylated NF-κB inhibitor (p-IκBα) (P<0.01) in the 75-day-old offspring rats. Compared with the model group, Zuoguiwan and vitamin E increased the primordial follicles in the offspring on the day of birth (P<0.01). Moreover, they resumed the estrous cycle, increased the ovary and uterine indices (P<0.05, P<0.01) and corpus luteum (P<0.01), reduced atretic follicles (P<0.01), elevated the serum levels of AMH and E2 (P<0.05, P<0.01), lowered the serum levels of LH, FSH, IL-1β, and TNF-α (P<0.05, P<0.01), and down-regulated the expression of TLR4, NF-κB p65, MyD88, and p-IκB-α (P<0.05, P<0.01) in the 75-day-old offspring. ConclusionZuoguiwan can improve the ovarian reserve in the offspring rat model of congenital kidney deficiency by regulating the TLR4/NF-κB p65 signaling pathway.
8.Youguiwan Reduces Airway Inflammation in COPD Rats with Syndrome of Kidney-Yang Deficiency by Inhibiting Leptin/JAK2/STAT3 Signaling Pathway
Lan ZHENG ; Zeyuan LUO ; Min XIAO ; Xiaocui JIANG ; Yuhao MENG ; Siyi CHEN ; Jing ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):17-26
ObjectiveTo observe the effect of Youguiwan on the leptin/Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in the lung tissue of the rat model of chronic obstructive pulmonary disease (COPD) due to kidney-Yang deficiency. MethodForty rats were modeled for COPD with the syndrome of kidney-Yang deficiency by intratracheal instillation of lipopolysaccharide on day 1 and day 14 and continuous fumigation for 6 weeks, during which hydrocortisone was injected intramuscularly at an interval of 3 days. The modeled rats were randomized into model, high- (11.7 g·kg-1), medium- (5.85 g·kg-1), and low-dose (2.93 g·kg-1) Youguiwan, and aminophylline (0.054 g·kg-1) group. In addition, 8 SD rats were set as the blank group. After the completion of modeling, the rats in each group were administrated with the corresponding drug by gavage for 28 consecutive days. After the last administration, samples were collected. A lung function analyzer was used to evaluate the lung function of rats. Enzyme-linked immunosorbent assay was employed to measure the levels of interleukin-17A (IL-17A), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the bronchoalveolar lavage fluid (BALF). Hematoxylin-eosin staining was employed to observe the pathological changes in the lung tissue, and Masson staining was employed to observe the deposition of blue collagen fibers around bronchi in the lung tissue and calculate the inflammation score. The immunofluorescence assay was employed to measure the protein content of collagen type Ⅰ (ColⅠ) and α-smooth muscle actin (α-SMA) in the bronchi. The protein and mRNA levels of leptin, IL-17A, JAK2, and STAT3 in the lung tissue were determined by Western blot and real-time fluorescence quantitative polymerase chain reaction, respectively. ResultCompared with the blank group, the model group showed decreased lung function (P<0.01), elevated levels of IL-6, IL-17A, and TNF-α in the BALF (P<0.01), and increased lung inflammation score, deposition of subcutaneous collagen fibers in the airway, and ColⅠ and α-SMA proteins (P<0.01). Furthermore, the modeling up-regulated the proteins and mRNA levels of leptin, IL-17A, JAK2, and STAT3 in the lung tissue (P<0.01) and enhanced the phosphorylation of JAK2 and STAT3 (P<0.01). Compared with the model group, high- and medium-dose Youguiwan improved the lung function, decreased the inflammation score, reduced collagen fiber deposition and ColⅠ and α-SMA proteins, lowered the levels of IL-6, IL-17A, and TNF-α in the BALF, down-regulated the mRNA and protein levels of leptin, JAK2, STAT3, and IL-17A, and weakened the phosphorylation of JAK2 and STAT3 (P<0.05, P<0.01). The aminophylline group had higher IL-17A and TNF-α levels than the high-dose Youguiwan group, lower IL-17A level than the medium and low-dose Youguiwan groups, and lower TNF-α level than the low-dose Youguiwan group. Compared with the aminophylline group, the high- and medium-dose Youguiwan groups showed reduced deposition of collagen fibers and protein levels of ColⅠ and α-SMA around the bronchi in the lung tissue (P<0.05, P<0.01), decreased inflammation score, and down-regulated protein and mRNA levels of leptin, JAK2, STAT3, and IL-17A in the lung tissue. ConclusionYouguiwan can prevent airway remodeling by inhibiting IL-17A to reduce inflammation and collagen deposition in COPD rats, which may be related to the inhibition of the leptin/JAK2/STAT3 signaling pathway.
9.Sishenwan Ameliorates Visceral Sensitivity in Rat Model of Diarrhea-predominant Irritable Bowel Syndrome (Spleen-kidney Yang Deficiency) by Regulating p38 MAPK/JNK/TRPV1 Pathway
Siqi LI ; Yunlian HU ; Chengxia SU ; Min XIAO ; Xiaocui JIANG ; Na WEN ; Qian ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(21):10-18
ObjectiveTo investigate the effect and possible mechanism of Sishenwan in ameliorating visceral sensitivity in the rat model of diarrhea-predominant irritable bowel syndrome (IBS-D) due to spleen-kidney Yang deficiency. MethodForty male SPF-grade rats were randomly assigned into five groups: blank control, model, low- (3.51 g·kg-1) and high-dose (7.02 g·kg-1) Sishenwan, and Peifikang (0.54 g·kg-1) groups. Except the blank control group, the other groups underwent maternal separation stress and Sennae Folium decoction gavage for the modeling of IBS-D due to spleen-kidney Yang deficiency. After corresponding drug interventions, the general conditions of the rats were observed, and the number of defecation pellets within 6 h and the minimum threshold of abdominal withdrawal reflex (AWR) were measured. Enzyme-linked immunosorbent assay (ELISA) was used to measure the serum levels of tumor necrosis factor (TNF)-α, gastrin (GAS), corticosterone (CORT), and adrenocorticotropic hormone (ACTH). Hematoxylin-eosin (HE) staining was employed to observe pathological changes in the colon tissue. Toluidine blue staining was used to assess mast cell degranulation in the colon tissue. Western blot was performed to determine the protein levels of p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK), transient receptor potential vanilloid 1 (TRPV1), and protease-activated receptor 2 (PAR2) in the colon tissue. Immunohistochemistry was employed to measure the protein level of TRPV1 in the colon tissue, and immunofluorescence was used to detect the positive expression of substance P (SP) and calcitonin gene-related peptide (CGRP) in the colon tissue. ResultCompared with the blank control group, the model group showed increased number of defecation pellets within 6 h (P<0.01), decreased minimum threshold of AWR (P<0.01), elevated serum TNF-α level (P<0.01), lowered levels of GAS, CORT, and ACTH (P<0.05, P<0.01), increased mast cell degranulation rate (P<0.01), increased positive expression of TRPV1, SP, and CGRP (P<0.05, P<0.01), and upregulated protein levels of p38 MAPK, JNK, TRPV1, and PAR2 (P<0.01). Compared with the model group, the high-dose Sishenwan group showed increased minimum threshold of AWR (P<0.01), reduced defecation frequency in both the high-dose Sishenwan and Peifikang groups (P<0.05, P<0.01), lowered TNF-α level (P<0.05, P<0.01), elevated levels of GAS, CORT, and ACTH (P<0.05, P<0.01), decreased mast cell degranulation rate (P<0.01), reduced positive expression of TRPV1, SP, and CGRP (P<0.05, P<0.01), and downregulated protein levels of p38 MAPK, JNK, TRPV1, and PAR2 (P<0.05, P<0.01). ConclusionSishenwan can ameliorate visceral sensitivity in the rat model of diarrhea-predominant irritable bowel syndrome due to spleen-kidney Yang deficiency by regulating the p38 MAPK/JNK/TRPV1 signaling pathway.
10.Preparation of Phenolic Acid-sodium Hyaluronate Copolymers and in vitro Antioxidant Activity Assessment
Xiao-Yue ZHANG ; Xiao-Na WANG ; Min JIANG ; Ting-Ting HAN ; Jin-Song GONG ; Qing-Na LI ; Su-Zhen YANG ; Jin-Song SHI
Progress in Biochemistry and Biophysics 2024;51(8):1950-1962
ObjectiveSodium hyaluronate (HA) was used as the research object to modify it with phenolic acid in order to obtain the molecular structure with better antioxidant activity or even new activity. MethodsIn this study, 5 kinds of phenolic acid-sodium hyaluronate was prepared by free radical-mediated grafting method, and the grafts with the highest grafting degree were selected to optimize the synthesis conditions. Then, grafts structure and physicochemical properties were analyzed. The grafts were characterized by IR, UV, 1H NMR, FESEM and TGA spectra. The in vitro antioxidant capacity of grafts was determined by the scavenging ability of DPPH·, ABTS+· and O2-·. ResultsAmong 5 kinds of phenolic acid-sodium hyaluronate, the grafting rate of ferulic acid-sodium hyaluronate copolymer (FA-HA) was highest , which was chosen as experimental sample in the following tests. Firstly, the reaction conditions were investigated and the highest grafting rate was (16.59±0.31) mg/g at the optimal preparation conditions. Then, FA-HA structure and physicochemical properties were analyzed. Data from UV, IR, 1H NMR analyses, TGA showed that FA were successfully grafted to HA. Compared with HA, the results of gel permeation chrematography (GPC) showed that the molecular mass distribution ofFA-HA copolymer decreased from 34.4 to 31.5 ku, but the uniformity of molecular distribution was improved. FESEM results showed that the structure of copolymer exhibited a closely connected lamellar structure with a relatively smooth surface. TGA results showed that thermal stability of FA-HA had a little decline. The antioxidant performance in vitro results showed that, during 0.25-10 g/L, FA-HA can eliminate (83.76±4.86)% DPPH·, (76.95±5.06)% ABTS+· and (83.08±2.51)% O2-· respectively at 10 g/L. which were higher than that of native HA and FA. ConclusionFA and HA were successfully grafted together by free radical grafting, and the grafted FA-HA had better antioxidant activity in vitro, which provided a theoretical basis for further research and development of phenolic acid-HA grafts.

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