1.Effects of Dihuang Yinzi on RAGE/ROS/Apoptosis Pathway in SH-SY5Y Cells Induced by Aβ1-42
Zhongyuan PIAO ; Yafen WEI ; Lin SONG ; Limei AO ; Yichan LU ; Zhuo JINAG ; Lifen YAO ; Yanli LIU
Journal of Guangzhou University of Traditional Chinese Medicine 2017;34(4):543-550
Objective To investigate the effects of Dihuang Yinzi (DY) on the receptor for advanced glycation end-products(RAGE)/reactive oxygen species(ROS)/apoptosis pathway in SH-SY5Y cells induced by amyloid-beta1-42 (Aβ1-42) oligomer. Methods Firstly, we adopted methyl thiazolyl tetrazolium(MTT) method to detect the cell vitality in fetal bovine serum (FBS) group, blank serum group, and low-, middle- and high- dose DY-containing serum groups, so as to confirm the optimal concentration and treatment time of DY-containing serum. Secondly, we applied MTT method to detect cell vitality and applied Annexin V/propidium iodide (PI) staining method to observe the apoptosis of SH-SY5Y cells treated with 0~20 μmol/L Aβ1-42 for 24 and 48 h, so as toconfirm the optimal concentration and treatment time of Aβ1-42 for establishing Alzheimer's disease (AD) model in vitro. Thirdly, MTT method was used for the detection of cell vitality, and Annexin V/PI staining method was used for detection of the apoptosis of SH-SY5Y cells in blank serum group, model group, western medicine control group and low-, middle-and high-dose DY-containing serum groups, and Dihydroethidium (DHE) method was used for the assay of ROS contents, so as to observe the effect of DY on the recovery of injured SH-SY5Y cells induced by Aβ1-42. Finally, we applied Western blot method to detect the expression level of RAGE in SH-SY5Y cells of blank group, model group and DY-containing serum group; after Aβ1-42-induced SH-SY5Y cells were transfected with RAGE gene, we adopted DHE staining method and Annexin V/PI staining method to detect ROS content and cell apoptotic rate in all of the above groups, so as to observe the effect of DY on SH-SY5Y cell apoptosis and RAGE expression. Results The cell vitalities were increased in low- and middle-dose DY-containing serum groups at 24 h (P < 0.05 or P < 0.01 compared with that in the blank serum group). The conditions for the establishment of AD model in vitro were as follows: the optimal concentration of Aβ1-42 was 5μmol/L, and the treatment time was 24 h. The cell vitalities were significantly enhanced, the cell apoptotic rate and ROS content were significantly lowered in Aβ1-42-induced SH-SY5Y cells of the medication groups(P <0.05 or P < 0.01 compared with those in the model group) , and the cell vitality was the highest and the cell apoptotic rate was the lowest in the middle-dose DY-containing serum group. The RAGE expression level was decreased in Aβ1-42-induced SH-SY5Y cells of the middle-dose DY-containing serum group(P < 0.05 compared with that in the model group) . ROS content and cell apoptotic rate were decreased in Aβ1-42-induced SH-SY5Y cells transfected with RAGE gene in the middle-dose DY-containing serum group (P<0.01). Conclusion DY may play an anti-oxidative role through inhibiting the production of ROS and cell apoptosis, thus to suppress RAGE protein and to achieve the preventive and therapeutic effect for AD.
2.Distribution pattern of traditional Chinese medicine syndromes and analysis of influencing factors in pancreatic cancer
Zhiyao SHI ; Shiyu WU ; Shaojian REN ; Yichan LIU ; Yijie YIN ; Yu GAO ; Xixing WANG
Journal of Clinical Hepatology 2025;41(3):528-535
ObjectiveTo investigate the influencing factors for traditional Chinese medicine (TCM) syndromes in pancreatic cancer by analyzing 608 cases, and to provide a theoretical reference for TCM syndrome differentiation and standardized treatment of pancreatic cancer. MethodsA total of 608 patients with a pathological or clinical diagnosis of pancreatic cancer who were admitted to Shanxi Institute of Traditional Chinese Medicine, The Affiliated Hospital of Shanxi University of Chinese Medicine, and Shanxi Provincial Hospital of Integrated Traditional Chinese and Western Medicine from January 2019 to December 2023 were enrolled, and TCM syndrome differentiation was performed. The chi-square test was used for comparison of categorical data between groups. The clinical data with statistical significance between groups were included in the regression analysis, and the unordered polytomous logistic regression model was used to investigate the influencing factors for the TCM syndrome of pancreatic cancer. ResultsFor the 608 patients with pancreatic cancer, common initial symptoms included abdominal pain (32.40%), abdominal distension (23.85%), fatigue (16.12%), and emaciation (10.03%), and the main clinical symptoms included poor appetite (75.97%), abdominal pain (67.27%), fatigue (61.84%), abdominal distension (57.40%), and emaciation (53.62%). There were significant differences between the patients with different TCM syndromes of pancreatic cancer in sex (χ2=62.823, P<0.001), disease duration (χ2=14.868, P=0.011), clinical stage (χ2=21.006, P<0.001), lymph node metastasis (χ2=2.205, P=0.032), surgery (χ2=38.008, P<0.001), chemotherapy (χ2=21.384, P<0.001), radiotherapy (χ2=17.510, P=0.004), and immunotherapy (χ2=18.573, P=0.002). The logistic regression analysis showed that male sex was a protective factor against Qi and blood deficiency syndrome (odds ratio [OR]=0.081, 95% confidence interval [CI]: 0.031 — 0.213, P<0.001), Qi stagnation and blood stasis syndrome (OR=0.100, 95%CI: 0.041 — 0.247, P<0.001), and syndrome of Yin deficiency with internal heat (OR=0.158, 95%CI: 0.057 — 0.444, P<0.001), while it was a risk factor for the syndrome of damp-heat accumulation (OR=2.378, 95%CI: 1.074 — 5.266, P=0.033); the course of the disease of<1 year was a protective factor against Qi and blood deficiency syndrome (OR=0.167, 95%CI: 0.073 — 0.383, P<0.001), syndrome of spleen-kidney Yang deficiency (OR=0.183, 95%CI: 0.089 — 0.378, P<0.001), and syndrome of Yin deficiency and internal heat (OR=0.164, 95%CI: 0.070 — 0.385, P<0.001); clinical stage Ⅰ/Ⅱ/Ⅲ was a risk factor for damp-heat accumulation (OR=2.793, 95%CI: 1.259 — 6.196, P=0.012) and Qi stagnation and blood stasis syndrome (OR=7.863, 95%CI: 2.808 — 22.020, P<0.001); lymph node metastasis was a risk factor for Qi and blood deficiency syndrome (OR=4.005, 95%CI: 1.477 — 10.861, P=0.006); surgical treatment was a risk factor for Qi and blood deficiency syndrome (OR=4.151, 95%CI: 1.916 — 8.995, P<0.001), syndrome of spleen-kidney yang deficiency (OR=5.352, 95%CI: 2.436 — 11.759, P<0.001), Qi stagnation and blood stasis syndrome (OR=2.334, 95%CI: 1.071 — 5.088, P=0.033), and syndrome of Yin deficiency and internal heat (OR=4.167, 95%CI: 1.789 — 9.707, P<0.001); chemotherapy was a protective factor against damp-heat accumulation (OR=0.188, 95%CI: 0.082 — 0.428, P<0.001); radiotherapy was a risk factor for damp-heat accumulation (OR=2.571, 95%CI: 1.151 — 5.746, P=0.021) and syndrome of Yin deficiency with internal heat (OR=8.384, 95%CI: 3.348 — 20.997, P<0.001); immunotherapy was a risk factor for Qi and blood deficiency syndrome (OR=2.114, 95%CI: 1.021 — 4.379, P=0.044). ConclusionSex, course of the disease, clinical stage, presence or absence of lymph node metastasis, surgery, chemotherapy, radiotherapy, and immunotherapy are the main influencing factors for the TCM syndrome of pancreatic cancer.