1.Evaluation of transdermal absorption of self-assembled nanoparticles of Huangqin decoction loaded with terbinafine
Chengying SHEN ; Yican HE ; Xiao CHENG ; Chaoying DU ; Hongyan MIN ; Baode SHEN ; Lingfei TONG
China Pharmacy 2026;37(2):180-185
OBJECTIVE To investigate the effect of Huangqin decoction (HQD)-based self-assembled nanoparticles (SAN) co-loaded with terbinafine (TBF) (TBF-HQD-SAN NPs) on the transdermal absorption of TBF. METHODS High-speed centrifugation combined with dialysis was used to separate HQD-SAN, and TBF-HQD-SAN NPs were obtained by loading TBF using the ultrasound magnetic stirring method; the particle size distribution, Zeta potential and polydispersity index (PDI) of the nanoparticle were characterized, and the encapsulation efficiency (EE) and drug loading (DL) of TBF were determined; using in vitro and in vivo transdermal experiments, the differences in transdermal performance between TBF-HQD-SAN NPs and TBF raw materials, as well as TBF and HQD-SAN physical mixture (TBF-HQD-SAN PM), were compared and analyzed. RESULTS TBF- HQD-SAN NPs were spherical with a particle size of (177.60±2.57) nm, a PDI of 0.197 4±0.007 9, and a Zeta potential of (-14.63±0.85) mV. The EE and DL of TBF were (99.49±0.71)% and (3.22±0.10)% , respectively. In vitro transdermal experiments, compared with TBF raw materials, the steady-state permeation rate (Jss) and skin retention of TBF-HQD-SAN NPs increased by 3.34 times and 27.56 times, respectively (P<0.05); compared with TBF-HQD-SAN PM, its Jss and skinretention were increased by 2.04 times and 7.44 times, respectively (P<0.05). In vivo transdermal experiments 69号) showed that, the area under the drug-time curve and the maximum concentration of TBF-HQD-SAN NPs increased by 2.13 times and 2.06 times respectively compared to TBF raw materials, and increased by 1.59 times and 1.65 times respectively compared to TBF-HQD-SAN PM (P<0.05). CONCLUSIONS TBF-HQD-SAN NPs can significantly enhance the in vitro and in vivo transdermal absorption efficiency and skin retention of TBF.
2.Intervention of Exercise Combined with Shenghui Tang in M1AChR Regulation of Mitochondrial Autophagy Improves Learning and Memory Ability of Alzheimer's Disease Model Rats
Xiaoxue TAO ; Min XIAO ; Kunyang TANG ; Bo WANG ; Chunchun ZHAO ; Rongxiang LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):120-130
ObjectiveTo make clear exercise combined with Shenghui Tang interferes in acetylcholine receptor (M1AChR) to improve mitochondrial autophagy and enhance cognition of Alzheimer's disease (AD) model rats through the adenylate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. MethodsForty-eight male SD rats of SPF grade were randomly divided into a blank group, a model group, a Shenghui Tang group (9.3 g·kg-1), an exercise group, an exercise + Shenghui Tang group (9.3 g·kg-1), and a rapamycin group (1.5 mg·kg-1). Except for the blank group, the rat model of AD was constructed by injecting amyloid beta (Aβ1-42) into hippocampus stereotaxically. The exercise group received treadmill exercise for 4 weeks, while the Shenghui Tang group received intragastric administration for 4 weeks, and the exercise + Shenghui Tang group received treadmill exercise and intragastric administration of Shenghui Tang for 4 weeks simultaneously. After the intervention, the Morris water maze test was used to detect the learning and memory ability. Spontaneous behavior was observed in the open field test. The pathological structure of hippocampal neurons was observed by NISSl staining. The expression level of M1AChR in hippocampus was detected by immunohistochemistry (IHC). The autophagy ultrastructure of hippocampal neurons was observed by transmission electron microscopy. The apoptosis rate was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). The expression of Beclin1 and microtubule-associated protein light chain 3β (LC3β) was detected by immunofluorescence (IF). The protein expression of M1AChR, AMPK, p-AMPK, mTOR, Beclin1, LC3β, and chelate 1 (SQSTM1/p62) in hippocampus was detected by Western blot. ResultsCompared with the blank group, the model group exhibited significantly increased platform escape latency on the fifth day (P<0.01) and significantly decreased activity distance in the target quadrant and times of crossing the platform (P<0.01). The total movement distance in the open field, the time of movement in the central area, and the average speed obviously decreased (P<0.05). The arrangement of nerve cells in hippocampus CA1 region was dispersed, and the numbers of Nissl bodies and M1AChR positive cells significantly decreased (P<0.01). The expression of TUNEL positive cells was significantly increased (P<0.01). The typical autophagic lysosomal structure decreased. The protein expression of M1AChR, p-AMPK/AMPK, p-mTOR/mTOR, Beclin1, LC3Ⅱ/Ⅰ in hippocampus was significantly decreased (P<0.01), and the protein expression of p62 was significantly increased (P<0.01). Compared with the model group, the exercise + Shenghui Tang group exhibited obviously improved space exploration and positioning navigation ability (P<0.05, P<0.01). The total movement distance in the open field, the time of movement in the central area, and the average speed of movement significantly increased (P<0.01). The number of Nissl bodies significantly increased (P<0.01). The number of M1AChR positive cells in hippocampus was significantly increased (P<0.01). The expression of TUNEL positive cells was significantly decreased (P<0.01). The protein expression of M1AChR, p-AMPK/AMPK, p-mTOR/mTOR, Beclin1, LC3Ⅱ/Ⅰ in hippocampus was significantly increased (P<0.01), while the protein expression of p62 was significantly decreased (P<0.01). Compared with the exercise + Shenghui Tang group, the Shenghui Tang group and the exercise group showed significantly increased platform escape latency on the fifth day (P<0.01) and obviously decreased activity distance in the target quadrant and times of crossing the platform (P<0.05, P<0.01). The total movement distance in the open field, the time of movement in the central area, and the average speed of movement significantly decreased (P<0.01). The number of Nissl bodies and the number of M1AChR positive cells significantly decreased (P<0.01). The expression of TUNEL positive cells was obviously increased (P<0.05). Ultrastructure of the hippocampal region showed decreased autophagy level. The protein expression of M1AChR, p-AMPK/AMPK, p-mTOR/mTOR, LC3Ⅱ/Ⅰ in the hippocampus was obviously decreased in the Shenghui Tang group (P<0.05, P<0.01), while the protein expression of p62 was significantly increased (P<0.01). In the exercise group, the protein expression of M1AChR, p-AMPK/AMPK, Beclin1, and LC3Ⅱ/Ⅰ was obviously decreased (P<0.05, P<0.01), while the protein expression of p-mTOR/mTOR and p62 was significantly increased (P<0.01). ConclusionExercise combined with traditional Chinese medicine can enhance the expression of M1AChR in the hippocampus of AD model rats, induce autophagy through the AMPK/mTOR signaling pathway, and improve the learning and memory ability of AD rats.
3.Mechanisms of Tianma Goutengyin in Alleviating Neuronal Injury in Vascular Dementia Model Rats by Inhibiting A1 Astrocyte Activation via Regulating TNF-α/STAT3/α1ACT Signaling Pathway
Xiaoyan WANG ; Min ZHAO ; Feng TIAN ; Min XIAO ; Nan QU ; Fugui LIU ; Chixiao LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):56-65
ObjectiveTo investigate the effects of Tianma Goutengyin on the tumor necrosis factor-α (TNF-α)/signal transducer and activator of transcription 3 (STAT3)/α1-antichymotrypsin C-terminal tail fragment (α1ACT) signaling pathway and A1-type astrocytes in a rat model of vascular dementia. MethodsSeventy-two male Sprague-Dawley rats were randomly divided into six groups (n=12 per group): Sham-operated group, model group, Tianma Goutengyin high-, medium-, and low-dose groups (5.13, 10.26, and 20.52 g·kg-1), and a nimodipine group (8.1 mg·kg-1). The vascular dementia model was established by permanent bilateral common carotid artery occlusion, followed by 4 weeks of intervention. Learning and memory ability were evaluated using the novel object recognition test, and behavioral performance was assessed using the forced swimming test. Levels of interleukin-6 (IL-6) and C-C motif chemokine ligand 2 (CCL2) in hippocampal tissue were measured by enzyme-linked immunosorbent assay (ELISA). Hippocampal neuronal morphology was observed by Nissl staining, and apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Immunohistochemistry was used to detect positive expression of brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP), and myelin basic protein (MBP). Western blot analysis was performed to measure the protein expression levels of TNF-α, TNF receptor 1 (TNFR1), phosphorylated STAT3 (p-STAT3), α1ACT, IL-6, complement component 3 (C3), BDNF, S100 calcium-binding protein A10 (S100A10), and GFAP in hippocampal tissue. ResultsCompared with the sham-operated group, the model group showed a significantly reduced relative recognition index in the novel object recognition test (P<0.01), prolonged immobility time and increased immobility frequency in the forced swimming test (P<0.01). Hippocampal IL-6 and CCL2 levels were significantly increased (P<0.01). Nissl staining revealed a marked reduction in neuronal number and loss of Nissl bodies (P<0.01). MBP-positive expression was significantly decreased (P<0.01), apoptosis was significantly increased (P<0.01), BDNF-positive expression was significantly reduced (P<0.05), and GFAP-positive expression was significantly increased (P<0.01). In addition, the protein expression levels of TNF-α, TNFR1, p-STAT3, α1ACT, IL-6, and C3 were significantly elevated (P<0.01), while BDNF and S100A10 expression levels were significantly decreased (P<0.01). Compared with the model group, all Tianma Gouteng yin dose groups exhibited a significant increase in the relative recognition index (P<0.05), shortened immobility time and reduced immobility frequency (P<0.05, P<0.01). IL-6 and CCL2 levels were significantly decreased (P<0.01), neuronal number was significantly increased (P<0.05, P<0.01), and MBP-positive expression was significantly enhanced (P<0.01). Apoptosis was significantly reduced (P<0.01), BDNF-positive expression was significantly increased (P<0.05), and GFAP-positive expression was significantly decreased (P<0.01). Moreover, the protein expression levels of TNF-α, TNFR1, p-STAT3, α1ACT, IL-6, and C3 were significantly decreased (P<0.01), while BDNF and S100A10 protein expression levels were significantly increased (P<0.01). ConclusionTianma Goutengyin may inhibit A1-type astrocyte activation in rats with vascular dementia through the TNF-α/STAT3/α1ACT signaling pathway, thereby reducing neuronal apoptosis and improving learning and memory function.
4.Tasquinimod promotes the sensitivity of ovarian cancer cells to cisplatin by down-regulating the HDAC4/p21 pathway
Zhao LI ; Ya-Hong WU ; Ye-Qing GUO ; Xiao-Jia MIN ; Ying LIN
The Korean Journal of Physiology and Pharmacology 2025;29(2):191-204
To investigate whether Tasquinimod can influence cisplatin resistance in drug-resistant ovarian cancer (OC) cell lines by regulating histone deacetylase 4 (HDAC4) or p21, we explored its effects on the cell cycle, and associated mechanisms.RT-PCR and Western blot analyses, flow cytometry, CCK8 assay, and immunofluorescence were utilized to investigate the effects of Tasquinimod on gene expression, cell cycle, apoptosis, viability, and protein levels in OC cells. The results showed that Tasquinimod inhibited cell viability and promoted apoptosis in SKOV3/DDP (cisplatin) and A2780/DDP cells more effectively than DDP alone. In combination with cisplatin, Tasquinimod further enhanced cell apoptosis and reduced cell viability in these cell lines, an effect that could be reversed following HDAC4 overexpression. Tasquinimod treatment down-regulated HDAC4, Bcl-2, and cyclin D1, and CDK4 expression and up-regulated the cleaved-Caspase-3, and p21 expression in SKOV3/DDP and A2780/ DDP cells. Additionally, Tasquinimod inhibited DDP resistance in OC/DDP cells. These effects were similarly observed in OC mouse models treated with Tasquinimod. In conclusion, Tasquinimod can improve OC cells' sensitivity to DDP by down-regulating the HDAC4/p21 axis, offering insights into potential strategies for overcoming cisplatin resistance in OC.
5.Rapid Identification of Different Parts of Nardostachys jatamansi Based on HS-SPME-GC-MS and Ultra-fast Gas Phase Electronic Nose
Tao WANG ; Xiaoqin ZHAO ; Yang WEN ; Momeimei QU ; Min LI ; Jing WEI ; Xiaoming BAO ; Ying LI ; Yuan LIU ; Xiao LUO ; Wenbing LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):182-191
ObjectiveTo establish a model that can quickly identify the aroma components in different parts of Nardostachys jatamansi, so as to provide a quality control basis for the market circulation and clinical use of N. jatamansi. MethodsHeadspace solid-phase microextraction-gas chromatography-mass spectrometry(HS-SPME-GC-MS) combined with Smart aroma database and National Institute of Standards and Technology(NIST) database were used to characterize the aroma components in different parts of N. jatamansi, and the aroma components were quantified according to relative response factor(RRF) and three internal standards, and the markers of aroma differences in different parts of N. jatamansi were identified by orthogonal partial least squares-discriminant analysis(OPLS-DA) and cluster thermal analysis based on variable importance in the projection(VIP) value >1 and P<0.01. The odor data of different parts of N. jatamansi were collected by Heracles Ⅱ Neo ultra-fast gas phase electronic nose, and the correlation between compound types of aroma components collected by the ultra-fast gas phase electronic nose and the detection results of HS-SPME-GC-MS was investigated by drawing odor fingerprints and odor response radargrams. Chromatographic peak information with distinguishing ability≥0.700 and peak area≥200 was selected as sensor data, and the rapid identification model of different parts of N. jatamansi was established by principal component analysis(PCA), discriminant factor alysis(DFA), soft independent modeling of class analogies(SIMCA) and statistical quality control analysis(SQCA). ResultsThe HS-SPME-GC-MS results showed that there were 28 common components in the underground and aboveground parts of N. jatamansi, of which 22 could be quantified and 12 significantly different components were screened out. Among these 12 components, the contents of five components(ethyl isovalerate, 2-pentylfuran, benzyl alcohol, nonanal and glacial acetic acid,) in the aboveground part of N. jatamansi were significantly higher than those in the underground part(P<0.01), the contents of β-ionone, patchouli alcohol, α-caryophyllene, linalyl butyrate, valencene, 1,8-cineole and p-cymene in the underground part of N. jatamansi were significantly higher than those in the aboveground part(P<0.01). Heracles Ⅱ Neo electronic nose results showed that the PCA discrimination index of the underground and aboveground parts of N. jatamansi was 82, and the contribution rates of the principal component factors were 99.94% and 99.89% when 2 and 3 principal components were extracted, respectively. The contribution rate of the discriminant factor 1 of the DFA model constructed on the basis of PCA was 100%, the validation score of the SIMCA model for discrimination of the two parts was 99, and SQCA could clearly distinguish different parts of N. jatamansi. ConclusionHS-SPME-GC-MS can clarify the differential markers of underground and aboveground parts of N. jatamansi. The four analytical models provided by Heracles Ⅱ Neo electronic nose(PCA, DFA, SIMCA and SQCA) can realize the rapid identification of different parts of N. jatamansi. Combining the two results, it is speculated that terpenes and carboxylic acids may be the main factors contributing to the difference in aroma between the underground and aboveground parts of N. jatamansi.
6.The level of HBV cccDNA in liver tissue and its clinical significance in patients in the convalescence stage of hepatitis B virus-related acute-on-chronic liver failure
Zhekai CAI ; Long XU ; Wenli LIU ; Yingqun XIAO ; Qingmei ZHONG ; Wei ZHANG ; Min WU
Journal of Clinical Hepatology 2025;41(1):57-62
ObjectiveTo investigate the expression level of HBV cccDNA in patients in the convalescence stage of hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) and its correlation with HBV markers and liver histopathological changes. MethodsA total of 30 patients in the convalescence stage of HBV-ACL who were hospitalized in The Ninth Hospital of Nanchang from January 2015 to October 2023 were enrolled as liver failure group, and 9 patients with chronic hepatitis B (CHB), matched for sex and age, were enrolled as control group. The content of HBV cccDNA in liver tissue was measured, and its correlation with clinical data and laboratory markers was analyzed. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and a one-way analysis of variance or the Kruskal-Wallis H test was used for comparison between multiple groups; the Fisher’s exact test was used for comparison of categorical data between groups. A Spearman correlation analysis was performed. ResultsThe liver failure group had a significantly lower content of HBV cccDNA in liver tissue than the control group (-0.92±0.70 log10 copies/cell vs -0.13±0.91 log10 copies/cell, t=2.761, P=0.009). In the liver failure group, there was no significant difference in the content of HBV cccDNA in liver tissue between the HBeAg-positive patients and the HBeAg-negative patients (P>0.05); there was no significant difference in the content of HBV cccDNA in liver tissue between the patients with different grades (G0-G2, G3, and G4) of liver inflammatory activity (P>0.05); there was no significant difference in the content of HBV cccDNA in liver tissue between the patients with different stages (S0-S2, S3, and S4) of liver fibrosis (P>0.05); there was no significant difference in the content of HBV cccDNA in liver tissue between the patients with negative HBV DNA and those with positive HBV DNA (P>0.05). For the liver failure group, the content of HBV cccDNA in liver tissue was positively correlated with the content of HBV DNA in liver tissue (r=0.426, P=0.043) and was not significantly correlated with the content of HBV DNA in serum (P>0.05). ConclusionThere is a significant reduction in the content of HBV cccDNA in liver tissue in the convalescence stage of HBV-ACLF. HBV cccDNA exists continuously and stably in liver tissue and can better reflect the persistent infection and replication of HBV than HBV DNA in serum and liver tissue.
7.AI Rudi's Experience in Diagnosing and Treating Skin Pruritus Based on the "Unity of Restoring Form, Regulating Qi,and Harmonizing Spirit"
Wei SONG ; Tianlin YANG ; Shuihan ZHOU ; Jie ZHANG ; Qianying YU ; Min XIAO ;
Journal of Traditional Chinese Medicine 2025;66(9):883-889
This paper summarizes Professor AI Rudi's experience in the diagnosis and treatment of skin pruritus based on the "unity of restoring form, regulating qi, and harmonizing spirit", employing internal herbal medicine combined with external treatments. It is believed that the core pathogenesis of pruritus is the "imbalance of form, qi, and spirit", with disturbed spirit as the onset, disordered qi as the key pathogenic factor, and physical changes as the manifestation of the disease. The treatment principle follows "restoring form-regulating qi-harmonizing spirit", with a combination of internal and external therapies, and differentiation based on deficiency and excess. For excess conditions caused by pathogenic disturbances to the heart spirit, treatment is based on different patterns of wind-heat, damp-heat, and blood-heat, using Sangye (Morus alba)-Sangbaipi (Morus alba cortex)-Longchi (Draconis os) to disperse wind and clear heat, calm the spirit; Difuzi (Kochia scoparia)-Qinghao (Artemisia annua)-Tanxiang (Santalum album) to clear damp-heat and aromatically open the spirit; Mudanpi (Paeonia suffruticosa)-Chuanxiong (Ligusticum chuanxiong)-Shuiniujiao (Bubalus bubalis cornua) to cool the blood, activate circulation, and calm the spirit. For deficiency conditions caused by insufficient nourishment of the heart spirit, treatment is based on patterns of qi deficiency or blood deficiency, using Huangqi (Astragalus membranaceus)-Fuping (Lemna minor)-Wuweizi (Schisandra chinensis) to tonify the qi and stabilize the exterior; Heshouwu (Polygonum multiflorum)-Jili (Tribulus terrestris)-Shouwuteng (Polygonum multiflorum vine) to nourish the blood, moisten dryness, and calm the spirit. External treatments integrate traditional Chinese medicine therapies such as medicinal baths, gua sha, and ear acupuncture, with custom herbal wash formulas for restoring form, jojoba oil gua sha for regulating qi, and ear seed therapy using Wangbuliuxing (Vaccaria segetalis) for harmonizing the spirit, achieving a holistic treatment effect for form, qi, and spirit.
8.The Role and Mechanism of Circadian Rhythm Regulation in Skin Tissue Regeneration
Ya-Qi ZHAO ; Lin-Lin ZHANG ; Xiao-Meng MA ; Zhen-Kai JIN ; Kun LI ; Min WANG
Progress in Biochemistry and Biophysics 2025;52(5):1165-1178
Circadian rhythm is an endogenous biological clock mechanism that enables organisms to adapt to the earth’s alternation of day and night. It plays a fundamental role in regulating physiological functions and behavioral patterns, such as sleep, feeding, hormone levels and body temperature. By aligning these processes with environmental changes, circadian rhythm plays a pivotal role in maintaining homeostasis and promoting optimal health. However, modern lifestyles, characterized by irregular work schedules and pervasive exposure to artificial light, have disrupted these rhythms for many individuals. Such disruptions have been linked to a variety of health problems, including sleep disorders, metabolic syndromes, cardiovascular diseases, and immune dysfunction, underscoring the critical role of circadian rhythm in human health. Among the numerous systems influenced by circadian rhythm, the skin—a multifunctional organ and the largest by surface area—is particularly noteworthy. As the body’s first line of defense against environmental insults such as UV radiation, pollutants, and pathogens, the skin is highly affected by changes in circadian rhythm. Circadian rhythm regulates multiple skin-related processes, including cyclic changes in cell proliferation, differentiation, and apoptosis, as well as DNA repair mechanisms and antioxidant defenses. For instance, studies have shown that keratinocyte proliferation peaks during the night, coinciding with reduced environmental stress, while DNA repair mechanisms are most active during the day to counteract UV-induced damage. This temporal coordination highlights the critical role of circadian rhythms in preserving skin integrity and function. Beyond maintaining homeostasis, circadian rhythm is also pivotal in the skin’s repair and regeneration processes following injury. Skin regeneration is a complex, multi-stage process involving hemostasis, inflammation, proliferation, and remodeling, all of which are influenced by circadian regulation. Key cellular activities, such as fibroblast migration, keratinocyte activation, and extracellular matrix remodeling, are modulated by the circadian clock, ensuring that repair processes occur with optimal efficiency. Additionally, circadian rhythm regulates the secretion of cytokines and growth factors, which are critical for coordinating cellular communication and orchestrating tissue regeneration. Disruptions to these rhythms can impair the repair process, leading to delayed wound healing, increased scarring, or chronic inflammatory conditions. The aim of this review is to synthesize recent information on the interactions between circadian rhythms and skin physiology, with a particular focus on skin tissue repair and regeneration. Molecular mechanisms of circadian regulation in skin cells, including the role of core clock genes such as Clock, Bmal1, Per and Cry. These genes control the expression of downstream effectors involved in cell cycle regulation, DNA repair, oxidative stress response and inflammatory pathways. By understanding how these mechanisms operate in healthy and diseased states, we can discover new insights into the temporal dynamics of skin regeneration. In addition, by exploring the therapeutic potential of circadian biology in enhancing skin repair and regeneration, strategies such as topical medications that can be applied in a time-limited manner, phototherapy that is synchronized with circadian rhythms, and pharmacological modulation of clock genes are expected to optimize clinical outcomes. Interventions based on the skin’s natural rhythms can provide a personalized and efficient approach to promote skin regeneration and recovery. This review not only introduces the important role of circadian rhythms in skin biology, but also provides a new idea for future innovative therapies and regenerative medicine based on circadian rhythms.
9.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.
10.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.

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