1.Yishen Huashi Granules Protect Kidneys of db/db Mice via p38 MAPK Signaling Pathway
Kaidong ZHOU ; Sitong WANG ; Ge JIN ; Yanmo CAI ; Xin ZHOU ; Yunhua LIU ; Xinxue ZHANG ; Min ZHANG ; Zongjiang ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):58-68
ObjectiveTo explore the mechanism of Yishen Huashi granules in alleviating renal tubular epithelial cell injury and relieving diabetic kidney disease by regulating the mitogen-activated protein kinase (MAPK) signaling pathway. MethodsThe db/db mice of 12 weeks old were randomly assigned into model , dapagliflozin (1.6 mg·kg-1), and Yishen Huashi granules (4.7 g·kg-1), and db/m mice were used as the control group. The general conditions of mice were observed, and fasting blood glucose and 24-h urinary protein and albumin-to-creatinine ratio (ACR) were measured at weeks 0 and 12 of administration. After 12 weeks of treatment, the levels of serum creatinine (SCr), blood urea (UREA), triglycerides (TG), total cholesterol (TC), and low density lipoprotein (LDL) were measured. The pathological changes in the renal tissue were observed by hematoxylin-eosin (HE) staining, Periodic acid-Schiff (PAS) staining, Mallory staining, and transmission electron microscopy. Real-time PCR was employed to determine the mRNA levels of monocyte chemotactic protein-1 (MCP-1) and CC chemokine receptor-2 (CCR2) in the renal tissue of mice. The immunohistochemical assay was employed to examine the expression of p38, phospho-p38 (p-p38), MCP-1, and CCR2 in the renal tissue of mice. Western blotting was employed to measure the protein levels of p-p38, p38, MCP-1, and CCR2 in the renal tissue of mice.HK-2 cells cultured in vitro were grouped as follows: negative control, high glucose(30 mmol·L-1), Yishen Huashi granule-containing serum, and SB203580. After 48 h of cell culture in each group, RNA were extracted and the levels of MCP-1, and CCR2 mRNA were determined by Real-time PCR,proteins were extracted and the levels of p38, p-p38, MCP-1, and CCR2 were determined by Western blot. ResultsThe in vivo experiments showed that before treatment, other groups had higher body weight, blood glucose level, 24 h urinary protein, and ACR than the control group (P<0.05,P<0.01). After 12 weeks of treatment, compared with the model group, the Yishen Huashi granules group showed improved general conditions, a decreasing trend in body weight, lowered levels of blood glucose, 24-h urinary protein, and ACR (P<0.01), reduced SCr and UREA (P<0.01), and declined levels of TC, TG, and LDL (P<0.05,P<0.01). Compared with the model group, the Yishen Huashi granules group showed alleviated damage and interstitial fibrosis in the renal tissue as well as reductions in glomerular foot process fusion and basement membrane thickening. Moreover, the Yishen Huashi granules group showed down-regulated mRNA levels of MCP-1 and CCR2 (P<0.01), reduced positive expression of p-p38, MCP-1, and CCR2 (P<0.01), and down-regulated protein levels of p-p38/p38, MCP-1, and CCR2 (P<0.05) in the renal tissue. The cell experiment showed that compared with the high glucose group, the Yishen Huashi granule-containing serum group showcased down-regulated mRNA levels of MCP-1 and CCR2 (P<0.01) and down-regulated protein levels of p-p38/p38, MCP-1, and CCR2(P<0.05,P<0.01). ConclusionYishen Huashi granules can regulate glucose-lipid metabolism, reduce 24 h urinary protein and ACR, improve the renal function, alleviate the renal tubule injury caused by high glucose, and protect renal tubule epithelial cells in db/db mice by reducing MCP-1/CCR2 activation via the p38 MAPK signaling pathway.
2.Efficacy and Mechanism of Action of Ermiao Situ Decoction in Modulating JAK/STAT Pathway in Rats with Damp-heat Eczema
Kangning HAN ; Junjie HU ; Juan LI ; Min ZHANG ; Xian ZHOU ; Songlin LIU ; Xin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):37-47
ObjectiveUltra performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) coupled with network pharmacology and molecular docking was utilized to explore the efficacy and mechanism of action of Ermiao Situ decoction on rats with damp-heat eczema. MethodsA rat model of damp-heat eczema was established by artificial climate chamber intervention combined with sensitization induction by dinitrochlorobenzene (DNCB), and it was randomly divided into the normal group, the model group, the medium- and high-dose groups of Ermiao Situ decoction (3.40 g·kg-1 and 6.80 g·kg-1), and the prednisone acetate group (2.51 mg·kg-1), with eight rats in each group, totalling 46 rats, of which six rats were tested with the drug-containing serum. The chemical analysis of drug-containing serum from rats was carried out by UPLC-Q-TOF-MS/MS, combined with network pharmacology for the prediction of key components, core targets, and signaling pathways, and molecular docking experiments were performed by CB-Dock2 online website. The pharmacological effects of Ermiao Situ decoction in the treatment of damp-heat eczema were investigated by epitaxial indexes combined with the pathologic tissue staining method. The serum levels of gastrin (GAS), interleukin-4 (IL-4), and interleukin-13 (IL-13) were measured by enzyme-linked immunosorbent assay (ELISA). Interleukin-6 (IL-6), Janus kinase 1 (JAK1), phosphorylated (p)-JAK1, signal transduction and activation of transcription factor 3 (STAT3), and p-STAT3 protein expression level was determined by Western bolt. ResultsA total of 19 active ingredients were detected in drug-containing serum samples of rats, which were predicted to act on 198 targets for the treatment of damp-heat eczema, among which the key ingredients included rhodopsin, huangpai alkaloids, and quercetin, and the main core targets included STAT3, tumor necrosis factor (TNF), and IL-6, which were mainly involved in the cancer signaling pathway, phosphatidylinositol 3-kinase (PI3K)/protein kinase (Akt) signaling pathway, T helper 17 (Th17) cell differentiation signaling pathway, and JAK/STAT signaling pathway. The molecular docking results suggested that the key components had strong binding activities with the core targets IL-6, JAK1, and STAT3 in the JAK/STAT signaling pathway. The results of animal experiments showed that compared with those in the normal group, rats in the model group were depressed. They had loose hair, loose stools, epidermal oozing, vesiculation, and generation of thick scabs in the form of scales, decreased body weight, increased anus temperature and water intake, and increased indexes of the spleen, thymus gland, and stomach (P<0.05, P<0.01), and the lesion tissue could be seen to be hyperkeratotic, with the aggregation of inflammatory cells and nonsignificant separation of epidermis and dermis. The gastric mucosa was thinned, deficient, and structurally disorganized, and obvious inflammatory cell aggregation was seen. The levels of GAS, IL-4, and IL-13 in serum were significantly reduced (P<0.05, P<0.01), and the protein expression levels of IL-6, JAK1, p-JAK1, and p-STAT3 in the lesion tissue were significantly increased (P<0.05, P<0.01). Compared with those in the model group, rats in each administration group had stable mental states, formed feces, a clean perianal area, and basically normal epidermis. Only a small amount of scaly scabs existed, and the rats had body weight increased, with decreased anal temperature and water intake, as well as decreased spleen, thymus, and gastric indexes (P<0.05, P<0.01). Epidermal thickness was decreased, and epidermal and dermal separation boundaries were obvious, but hyperkeratotic and accumulation of inflammatory cells could still be seen. The thickness of gastric mucosa increased, and the structure was restored to varying degrees. The levels of GAS, IL-4, and IL-13 content in the serum of rats were increased to varying degrees, and the protein expression levels of IL-6, JAK1, p-JAK1, and p-STAT3 in the dermal lesion tissue were significantly decreased (P<0.05, P<0.01). ConclusionErmiao Situ decoction may exert therapeutic effects on rats with damp-heat eczema by modulating the JAK/STAT signaling pathway.
3.Process Optimization and Health Risk Assessment of Calcined Haematitum Based on QbD Concept
Yue YANG ; Jingwei ZHOU ; Jialiang ZOU ; Guorong MEI ; Yifan SHI ; Lei ZHONG ; Jiaojiao WANG ; Xuelian GAN ; Dewen ZENG ; Xin CHEN ; Lin CHEN ; Hongping CHEN ; Shilin CHEN ; Yuan HU ; Youping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):187-196
ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design(QbD) and to assess its health risk. MethodsTaking whole iron content, Fe2+ dissolution content and looseness as critical quality attributes(CQAs), and calcination temperature, calcination time, spreading thickness and particle size as critical process parameters(CPPs) determined by the failure mode and effect analysis(FMEA), the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation(AHP-CRITIC) hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry, and the health risk assessment was carried out by daily exposure(EXP), target hazard quotient(THQ) and lifetime cancer risk(LCR), and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃, calcination time of 1 h, particle size of 0.2-0.5 cm, spreading thickness of 1 cm, and vinegar quenching for 1 time[Haematitum-vinegar(10:3)]. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1, while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10-6-1×10-4, and the LCR of the rest was<1×10-6, and the LCRs of calcined Haematitum were all<1×10-6, indicating that the carcinogenic risk of calcined Haematitum was low, but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu, As, Cd, Pb and Hg were formulated as 1 014, 25, 17, 27, 7 mg·kg-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible, and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu, As, Cd, Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.
4.Optimization of Processing Technology of Calcined Pyritum Based on QbD Concept and Its XRD Fingerprint Analysis
Xin CHEN ; Jingwei ZHOU ; Haiying GOU ; Lei ZHONG ; Tianxing HE ; Wenbo FEI ; Jialiang ZOU ; Yue YANG ; Dewen ZENG ; Lin CHEN ; Hongping CHEN ; Shilin CHEN ; Yuan HU ; Youping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):197-205
ObjectiveBased on the concept of quality by design(QbD), the processing process of calcined Pyritum was optimized, and its X-ray diffraction(XRD) fingerprint was established. MethodsThe safety, effectiveness and quality controllability of calcined Pyritum were taken as the quality profile(QTPP), the color, hardness, metallic luster, phase composition, the contents of heavy metals and hazardous elements were taken as the critical quality attributes(CQAs), and the calcination temperature, calcination time, paving thickness and particle size were determined as the critical process parameters(CPPs). Differential thermal analysis, X-ray diffraction(XRD) and inductively coupled plasma mass spectrometry(ICP-MS) were used to analyze the correlation between the calcination temperature and CQAs of calcined Pyritum. Then, based on the criteria importance through intercriteria correlation(CRITIC)-entropy weight method, the optimal processing process of calcined Pyritum was optimized by orthogonal test. Powder XRD was used to analyze the phase of calcined Pyritum samples processed according to the best process, and the mean and median maps of calcined Pyritum were established by the superposition of geometric topological figures, and similarity evaluation and cluster analysis were carried out. ResultsThe results of single factor experiments showed that the physical phase of Pyritum changed from FeS2 to Fe7S8 during the process of temperature increase, the color gradually deepened from dark yellow, and the contents of heavy metals and harmful elements decreased. The optimized processing process of calcined Pyritum was as follows:calcination temperature at 750 ℃, calcination time of 2.5 h, paving thickness of 3 cm, particle size of 0.8-1.2 cm, vinegar quenching 1 time[Pyritum-vinegar(10∶3)]. After calcination, the internal structure of Pyritum was honeycomb-shaped, which was conducive to the dissolution of active ingredients. XRD fingerprints of 13 batches of calcined Pyritum characterized by 10 common peaks were established. The similarities of the relative peak intensities of the XRD fingerprints of the analyzed samples were>0.96, and it could effectively distinguish the raw products and unqualified products. ConclusionTemperature is the main factor affecting the quality of calcined Pyritum. After processing, the dissolution of the effective components in Pyritum increases, and the contents of heavy metals and harmful substances decrease, reflecting the function of processing to increase efficiency and reduce toxicity. The optimized processing process is stable and feasible, and the established XRD fingerprint can be used as one of the quality control standards of calcined Pyritum.
5.Effect of Shenkang Injection on Podocyte Apoptosis and GRP78/CHOP Signaling Pathway in db/db Mice with Diabetic Kidney Disease Based on Endoplasmic Reticulum Stress
Yanmo CAI ; Sitong WANG ; Xin ZHOU ; Ge JIN ; Kaidong ZHOU ; Yunhua LIU ; Fengfeng ZHANG ; Xinxue ZHANG ; Zongjiang ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):81-90
ObjectiveTo investigate the mechanism of Shenkang injection in delaying diabetic kidney disease by regulating endoplasmic reticulum stress and attenuating podocyte apoptosis through the Glucose regulated protein 78 ( GRP78 ) / transcription factor C / EBP homologous protein ( CHOP ) signaling pathway (GRP78/CHOP) signaling pathway. MethodsFor the animal experiment, 10 12-week-old db/m mice were selected as a normal group, and 30 12-week-old db/db mice were randomly divided into a model group, a Shenkang injection group (15.6 mL·kg-1), and a dapagliflozin group (1.6 mg·kg-1). To observe the general condition of mice, fasting blood glucose, urinary albumin/urine creatinine (ACR), and 24 h urine protein quantification were detected in each group before drug administration. After 12 weeks of drug treatment, mice were tested for fasting blood glucose, total cholesterol (TC), triglyceride (TG), low-density cholesterol (LDL), ACR, 24 h urine protein quantification, blood creatinine (SCr), and blood urea (UREA). Hematoxylin-eosin (HE) staining, periodic acid-Schiff (PAS) staining, and transmission electron microscopy were used to observe the pathologic morphology in renal tissue. Immunohistochemistry was used to detect the expressions of nephroprotective marker protein (Nephrin), glucose-regulated protein 78 (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in renal tissue. Western blot was used to detect the expressions of GRP78, CHOP, Bcl-2, Bax, and Nephrin proteins, and Real-time polymerase chain reaction (Real-time PCR) was employed to detect the expressions of Nephrin, GRP78, CHOP, Bcl-2, and Bax mRNAs in renal tissue. ResultsBefore drug administration, compared with those in the normal group, the body mass of db/db mice was significantly increased, and blood glucose, 24 h urine protein quantification, and ACR were significantly elevated in the Shenkang injection group and Dapagliflozin group (P<0.01). After 12 weeks of administration, compared with those in the model group, the general state of mice in the Shenkang injection group was significantly improved, and the body mass was decreased. The blood glucose was significantly reduced (P<0.01), and blood lipids TC, TG, and LDL were significantly decreased (P<0.05, P<0.01). The 24 h urine protein quantification and ACR were significantly decreased (P<0.05), and SCr and UREA were significantly reduced (P<0.01). Compared with those of the model group, the pathologic results of the Shenkang injection group showed that proliferation of mesangial cells, reduction of inflammatory cell infiltration, and alleviation of renal tubular vacuolization and podocyte damage were observed in renal tissue of mice. Electron microscopy showed that fusion of the pedicle protruding and thickening of the basement membrane were reduced. Immunohistochemistry results showed that the expressions of GRP78, CHOP, and Bax proteins were significantly reduced (P<0.01), and the expressions of Nephrin and Bcl-2 proteins were significantly increased (P<0.01) in renal tissue of the Shenkang injection group. Western blot results showed that the expressions of Nephrin and Bcl-2 in the Shenkang injection group were significantly increased (P<0.05, P<0.01), and the expressions of GRP78, CHOP, and Bax proteins were significantly decreased (P<0.05, P<0.01). Real-time PCR results showed that the expressions of GRP78, CHOP, and Bax mRNAs were down regulated in the Shenkang injection group (P<0.01), and the expressions of Nephrin and Bcl-2 mRNAs were up regulated (P<0.01). ConclusionShenkang injection inhibits endoplasmic reticulum stress response and podocyte apoptosis by regulating the GRP78/CHOP signaling pathway, which in turn ensures the integrity of glomerular filtration barrier, reduces the occurrence of proteinuria, improves renal function, and thus delays the progression of diabetic kidney disease.
6.Process Optimization and Health Risk Assessment of Calcined Haematitum Based on QbD Concept
Yue YANG ; Jingwei ZHOU ; Jialiang ZOU ; Guorong MEI ; Yifan SHI ; Lei ZHONG ; Jiaojiao WANG ; Xuelian GAN ; Dewen ZENG ; Xin CHEN ; Lin CHEN ; Hongping CHEN ; Shilin CHEN ; Yuan HU ; Youping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):187-196
ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design(QbD) and to assess its health risk. MethodsTaking whole iron content, Fe2+ dissolution content and looseness as critical quality attributes(CQAs), and calcination temperature, calcination time, spreading thickness and particle size as critical process parameters(CPPs) determined by the failure mode and effect analysis(FMEA), the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation(AHP-CRITIC) hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry, and the health risk assessment was carried out by daily exposure(EXP), target hazard quotient(THQ) and lifetime cancer risk(LCR), and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃, calcination time of 1 h, particle size of 0.2-0.5 cm, spreading thickness of 1 cm, and vinegar quenching for 1 time[Haematitum-vinegar(10:3)]. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1, while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10-6-1×10-4, and the LCR of the rest was<1×10-6, and the LCRs of calcined Haematitum were all<1×10-6, indicating that the carcinogenic risk of calcined Haematitum was low, but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu, As, Cd, Pb and Hg were formulated as 1 014, 25, 17, 27, 7 mg·kg-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible, and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu, As, Cd, Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.
7.Optimization of Processing Technology of Calcined Pyritum Based on QbD Concept and Its XRD Fingerprint Analysis
Xin CHEN ; Jingwei ZHOU ; Haiying GOU ; Lei ZHONG ; Tianxing HE ; Wenbo FEI ; Jialiang ZOU ; Yue YANG ; Dewen ZENG ; Lin CHEN ; Hongping CHEN ; Shilin CHEN ; Yuan HU ; Youping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):197-205
ObjectiveBased on the concept of quality by design(QbD), the processing process of calcined Pyritum was optimized, and its X-ray diffraction(XRD) fingerprint was established. MethodsThe safety, effectiveness and quality controllability of calcined Pyritum were taken as the quality profile(QTPP), the color, hardness, metallic luster, phase composition, the contents of heavy metals and hazardous elements were taken as the critical quality attributes(CQAs), and the calcination temperature, calcination time, paving thickness and particle size were determined as the critical process parameters(CPPs). Differential thermal analysis, X-ray diffraction(XRD) and inductively coupled plasma mass spectrometry(ICP-MS) were used to analyze the correlation between the calcination temperature and CQAs of calcined Pyritum. Then, based on the criteria importance through intercriteria correlation(CRITIC)-entropy weight method, the optimal processing process of calcined Pyritum was optimized by orthogonal test. Powder XRD was used to analyze the phase of calcined Pyritum samples processed according to the best process, and the mean and median maps of calcined Pyritum were established by the superposition of geometric topological figures, and similarity evaluation and cluster analysis were carried out. ResultsThe results of single factor experiments showed that the physical phase of Pyritum changed from FeS2 to Fe7S8 during the process of temperature increase, the color gradually deepened from dark yellow, and the contents of heavy metals and harmful elements decreased. The optimized processing process of calcined Pyritum was as follows:calcination temperature at 750 ℃, calcination time of 2.5 h, paving thickness of 3 cm, particle size of 0.8-1.2 cm, vinegar quenching 1 time[Pyritum-vinegar(10∶3)]. After calcination, the internal structure of Pyritum was honeycomb-shaped, which was conducive to the dissolution of active ingredients. XRD fingerprints of 13 batches of calcined Pyritum characterized by 10 common peaks were established. The similarities of the relative peak intensities of the XRD fingerprints of the analyzed samples were>0.96, and it could effectively distinguish the raw products and unqualified products. ConclusionTemperature is the main factor affecting the quality of calcined Pyritum. After processing, the dissolution of the effective components in Pyritum increases, and the contents of heavy metals and harmful substances decrease, reflecting the function of processing to increase efficiency and reduce toxicity. The optimized processing process is stable and feasible, and the established XRD fingerprint can be used as one of the quality control standards of calcined Pyritum.
8.Effect of Shenkang Injection on Podocyte Apoptosis and GRP78/CHOP Signaling Pathway in db/db Mice with Diabetic Kidney Disease Based on Endoplasmic Reticulum Stress
Yanmo CAI ; Sitong WANG ; Xin ZHOU ; Ge JIN ; Kaidong ZHOU ; Yunhua LIU ; Fengfeng ZHANG ; Xinxue ZHANG ; Zongjiang ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):81-90
ObjectiveTo investigate the mechanism of Shenkang injection in delaying diabetic kidney disease by regulating endoplasmic reticulum stress and attenuating podocyte apoptosis through the Glucose regulated protein 78 ( GRP78 ) / transcription factor C / EBP homologous protein ( CHOP ) signaling pathway (GRP78/CHOP) signaling pathway. MethodsFor the animal experiment, 10 12-week-old db/m mice were selected as a normal group, and 30 12-week-old db/db mice were randomly divided into a model group, a Shenkang injection group (15.6 mL·kg-1), and a dapagliflozin group (1.6 mg·kg-1). To observe the general condition of mice, fasting blood glucose, urinary albumin/urine creatinine (ACR), and 24 h urine protein quantification were detected in each group before drug administration. After 12 weeks of drug treatment, mice were tested for fasting blood glucose, total cholesterol (TC), triglyceride (TG), low-density cholesterol (LDL), ACR, 24 h urine protein quantification, blood creatinine (SCr), and blood urea (UREA). Hematoxylin-eosin (HE) staining, periodic acid-Schiff (PAS) staining, and transmission electron microscopy were used to observe the pathologic morphology in renal tissue. Immunohistochemistry was used to detect the expressions of nephroprotective marker protein (Nephrin), glucose-regulated protein 78 (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in renal tissue. Western blot was used to detect the expressions of GRP78, CHOP, Bcl-2, Bax, and Nephrin proteins, and Real-time polymerase chain reaction (Real-time PCR) was employed to detect the expressions of Nephrin, GRP78, CHOP, Bcl-2, and Bax mRNAs in renal tissue. ResultsBefore drug administration, compared with those in the normal group, the body mass of db/db mice was significantly increased, and blood glucose, 24 h urine protein quantification, and ACR were significantly elevated in the Shenkang injection group and Dapagliflozin group (P<0.01). After 12 weeks of administration, compared with those in the model group, the general state of mice in the Shenkang injection group was significantly improved, and the body mass was decreased. The blood glucose was significantly reduced (P<0.01), and blood lipids TC, TG, and LDL were significantly decreased (P<0.05, P<0.01). The 24 h urine protein quantification and ACR were significantly decreased (P<0.05), and SCr and UREA were significantly reduced (P<0.01). Compared with those of the model group, the pathologic results of the Shenkang injection group showed that proliferation of mesangial cells, reduction of inflammatory cell infiltration, and alleviation of renal tubular vacuolization and podocyte damage were observed in renal tissue of mice. Electron microscopy showed that fusion of the pedicle protruding and thickening of the basement membrane were reduced. Immunohistochemistry results showed that the expressions of GRP78, CHOP, and Bax proteins were significantly reduced (P<0.01), and the expressions of Nephrin and Bcl-2 proteins were significantly increased (P<0.01) in renal tissue of the Shenkang injection group. Western blot results showed that the expressions of Nephrin and Bcl-2 in the Shenkang injection group were significantly increased (P<0.05, P<0.01), and the expressions of GRP78, CHOP, and Bax proteins were significantly decreased (P<0.05, P<0.01). Real-time PCR results showed that the expressions of GRP78, CHOP, and Bax mRNAs were down regulated in the Shenkang injection group (P<0.01), and the expressions of Nephrin and Bcl-2 mRNAs were up regulated (P<0.01). ConclusionShenkang injection inhibits endoplasmic reticulum stress response and podocyte apoptosis by regulating the GRP78/CHOP signaling pathway, which in turn ensures the integrity of glomerular filtration barrier, reduces the occurrence of proteinuria, improves renal function, and thus delays the progression of diabetic kidney disease.
9.Exploration of the antidepressant machanism of Shugan hewei tang based on metabolomics of PFC-NAc-VTA neural circuit
Xinyue QU ; Junjie HU ; Juan LI ; Min ZHANG ; Xian ZHOU ; Songlin LIU ; Xin CHEN
China Pharmacy 2025;36(10):1172-1178
OBJECTIVE To investigate the antidepressant mechanism of Shugan hewei tang (SGHWT) based on the metabolomics of prefrontal cortex (PFC)-nucleus accumbens (NAc)-ventral tegmental area (VTA) neural circuit. METHODS Male SD rats were randomly divided into blank group, model group, SGHWT low-, medium- and high-dose groups [3.67, 7.34, 14.68 g/(kg·d), by raw material], and fluoxetine group [1.58 mg/(kg·d), positive control], with 12 rats in each group. Except for the blank group, the depression model was established by chronic unpredictable mild stress combined with individual cage housing in the remaining groups, and the corresponding drug solution or normal saline was administered via gavage during modeling, once a day, for 6 consecutive weeks. After the last administration, the body weight, sucrose preference rate, total moving distance, frequency into the center and immobility time of rats in each group were detected. Samples of PFC, NAc and VTA areas of rats in the blank group, model group, SGHWT medium-dose group and fluoxetine positive control groups were collected,and their histomorphological features were observed, and non-targeted metabolomics analysis (except for fluoxetine group)were performed and validated. RESULTS Compared with model group, the cytolysis, structural damage and other pathological damages in three brain regions of rats were significantly alleviated in each drug group, while their body weight, sucrose preference rate, total moving distance and frequency into the center were all significantly higher or longer (P<0.05), and immobility time was significantly shorter (P<0.05). The results of non-targeted metabolomics showed that a total of 78 endogenous differential metabolites were identified, with 40, 35 and 24 in the PFC, NAc and VTA regions respectively, mainly involved in amino acid, lipid and sphingolipid metabolism. The results of metabolic pathway enrichment analysis showed that SGHWT affected the neural circuits of depressed rats by regulating sphingolipid metabolism, alanine, aspartic acid and glutamic acid metabolism, saturated fatty acid biosynthesis, among which alanine, aspartic acid and glutamic acid metabolism was predominantly involved. Validation experiments showed that SGHWT significantly increased the phosphorylation levels of protein kinase B (Akt) and mammalian target of rapamycin (mTOR), and decreased the protein expression of N-methyl-D-aspartic acid receptor 1 (NMDAR1) in the NAc region of rats. CONCLUSIONS SGHWT significantly improves the depression-like behavior and attenuates pathological damage of PFC-NAc-VTA neural circuit of model rats, the mechanism of which is associated with inhibiting NMDAR1 expression and activating the Akt/mTOR signaling pathway.
10.Clinical Diseases Responding Specially to TCM Treatment: Psoriasis
Liu LIU ; Xiaoying SUN ; Mei MO ; Yaqiong ZHOU ; Bin LI ; Xiaoxiao ZHANG ; Xin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):260-268
Psoriasis is a common chronic inflammatory systemic disease in dermatology. Its high prevalence, recurrence rate, and numerous comorbidities impose a significant physical and mental burden on patients. With the continuous advancement of modern medicine, the emergence of biological agents has improved clinical efficacy, making it possible to overcome psoriasis, in addition to classical treatments. However, in clinical practice, adverse reactions, drug resistance, recurrence rates, and immune drift cannot be ignored. Traditional Chinese medicine (TCM) has a history of thousands of years in treating psoriasis, demonstrating good efficacy, high safety, and a low recurrence rate, but a standardized management system is lacking. Therefore, the 25th Clinical Diseases Responding Specially to TCM Treatment Series (Psoriasis) Youth Salon, hosted by the Chinese Association of Chinese Medicine and organized by the Youth Committee of the Chinese Association of Chinese Medicine, invited 29 experts and scholars from TCM, Western medicine, and interdisciplinary fields to actively discuss the "Advantages, Challenges, and Clinical Transformation of TCM and Western Medicine in the Diagnosis and Treatment of Psoriasis". The experts at the meeting concluded that the advantages of TCM in the treatment of psoriasis are as follows. Firstly, in the TCM-led treatment plan, TCM's understanding of psoriasis follows the principle of combining the differentiation of disease and syndrome. This approach distinguishes the basic contradiction from the current main contradiction and enables a clear grasp of the dynamic process of psoriasis development. Based on the system of syndrome differentiation and treatment, TCM intervention is applied to address the current main contradiction, and the optimal TCM treatment plan is formulated by combining internal and external treatments. Adhering to the principle of "what is visible outside must be addressed inside", TCM can prevent and treat psoriasis comorbidities early by differentiating syndrome types. Secondly, in the integrated TCM and Western medicine treatment plan, the combination of both methods not only enhances efficacy but also reduces the adverse reactions of immunosuppressants and biological agents, lowering the recurrence rate. This conference provides a reference for the diagnosis and treatment of psoriasis using TCM and integrated TCM and Western medicine, opening up new ideas for clinical and basic research and guiding future research directions.

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