1.Activation of Nrf2/HO-1/NQO1 Signaling Pathway by Shenqi Tangluo Pill Improves Oxidative Stress Injury of Skeletal Muscle of Type 2 Diabetes Mellitus Mice
Xiaoli PEI ; Yonglin LIANG ; ⁎ ; Yongqiang DUAN ; ⁎ ; Xiangdong ZHU ; Bing SONG ; Min BAI ; Yunhui ZHAO ; Sichen ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(7):131-139
ObjectiveTo investigate the effect and mechanism of Shenqi Tangluo pill (SQTLP) on oxidative stress injury of skeletal muscle of type 2 diabetes mellitus (T2DM) mice based on nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1)/NAD(P)H quinone oxidoreductase 1 (NQO1) pathway. MethodA total of 60 7-week-old male db/db mice [specific pathogen-free (SPF) grade] were selected and fed for one week for adaption. They were divided into the model control group, SQTLP low-, medium- and high-dose (19, 38, and 76 g·kg-1) groups and metformin group (0.26 g·kg-1) by gavage. Each group consisted of 12 mice. Twelve male db/m mice of the same age were selected as the blank group. The intervention was implemented continuously for 8 weeks. Fasting blood glucose (FBG) was detected. Fasting serum insulin (FINS) levels were detected by enzyme-linked immunosorbent assay (ELISA), and the homeostasis model assessment-insulin resistance (HOMA-IR) index and the homeostasis model assessment-insulin sensitivity index (HOMA-ISI) were calculated. Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were conducted. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and the contents of malondialdehyde (MDA) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) in skeletal muscle tissues were detected by biochemical kits. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in skeletal muscle tissues. The levels of reactive oxygen species (ROS) and 4-hydroxynonenal (4-HNE) in skeletal muscle tissue were detected by immunofluorescence (IF). The expression levels of Nrf2, HO-1, NQO1 and glutamate-cysteine ligase catalytic subunit (GCLC) proteins in skeletal muscle tissues were detected by Western blot. ResultCompared with those in the blank group, FBG, FINS and HOMA-IR in the model group were significantly increased (P<0.05), while HOMA-ISI was decreased (P<0.05). The results of OGTT and ITT showed that blood glucose was significantly increased at all time points (P<0.05), and glucose tolerance and insulin tolerance were significantly impaired. SOD and GSH-Px activities in skeletal muscle tissues were significantly decreased (P<0.05), and MDA and NADPH contents were significantly increased (P<0.05). In skeletal muscle tissues, the arrangement of muscle fibers was loose, the nucleus was disordered, and inflammatory cells were infiltrated. The expression levels of ROS and 4-HNE in skeletal muscle tissues were significantly increased (P<0.05). The protein expression levels of Nrf2, HO-1, NQO1 and GCLC in skeletal muscle tissues were significantly decreased (P<0.05). Compared with those in the model group, FBG, FINS and HOMA-IR in the metformin group were significantly decreased (P<0.05), while HOMA-ISI was increased (P<0.05). The results of OGTT and ITT showed that blood glucose in the metformin group was significantly decreased at all time points (P<0.05). The activities of SOD and GSH-Px in skeletal muscle tissues were significantly increased (P<0.05), while the contents of MDA and NADPH were significantly decreased (P<0.05). No obvious abnormality was found in the skeletal muscle tissue of the metformin group. The expressions of ROS and 4-HNE in skeletal muscle tissues were decreased (P<0.05). The protein expression levels of Nrf2, HO-1, NQO1 and GCLC in skeletal muscle tissues were significantly increased (P<0.05). Compared with those in the model group, FBG, FINS and HOMA-IR in the SQTLP medium- and high-dose groups were significantly decreased (P<0.05), while HOMA-ISI was increased (P<0.05). The results of OGTT and ITT showed that the glucose tolerance and insulin tolerance of mice were improved in each dose group of SQTLP. The GSH-Px activity in the SQTLP low-dose group was significantly increased (P<0.05), and the NADPH content was decreased (P<0.05). The activities of SOD and GSH-Px in the SQTLP medium- and high-dose groups were significantly increased (P<0.05), while the contents of MDA and NADPH were significantly decreased (P<0.05). The skeletal muscle tissue injury of mice in each dose group of SQTLP was ameliorated to different degrees. In the SQTLP medium- and high-dose groups, the expressions of ROS and 4-HNE were decreased (P<0.05), and the protein expression levels of Nrf2, HO-1, NQO1 and GCLC were significantly increased (P<0.05). Compared with those in the SQTLP low-dose group, FBG and HOMA-IR in the SQTLP high-dose group were significantly decreased (P<0.05), while HOMA-ISI was increased (P<0.05). The results of OGTT and ITT showed that the SQTLP high-dose group significantly improved the glucose tolerance and insulin tolerance of mice. The activities of SOD and GSH-Px in skeletal muscle tissues were significantly increased (P<0.05), while the contents of MDA and NADPH were significantly decreased (P<0.05). No obvious abnormality was found in the skeletal muscle tissue, the expressions of ROS and 4-HNE were decreased (P<0.05), and the protein expression levels of Nrf2, HO-1, NQO1 and GCLC were significantly increased (P<0.05) in the skeletal muscle tissue of the SQTLP high-dose group. ConclusionSQTLP can significantly improve IR in T2DM mice, and the mechanism is related to SQTLP activating the Nrf2/HO-1/NQO1 signaling pathway, promoting the expression of antioxidant enzymes, and thus improving the oxidative stress injury in the skeletal muscle.
2.Mechanism of Dahuang Tangluo Pills in Improving Renal Inflammatory Injury in Diabetic Kidkdey Disease by Regulating AGEs/RAGE/IKK/NF-κB Pathway
Pu ZHANG ; Jianqing LIANG ; Xia YANG ; Min BAI ; Xiangdong ZHU ; Chunxia XUE ; Beibei SU ; Yunhui ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(20):77-85
ObjectiveTo explore the protective effects of Dahuang Tangluo pills on early diabetic kidkdey disease (DKD) in db/db mice. MethodEight db/m mice were selected as the control group. Forty male db/db mice were selected and blood samples were collected via tail vein to measure fasting blood glucose (FBG). Mice with FBG ≥ 16.7 mmol·L-1, increased urine output, and persistent albuminuria were considered successful in model establishment. After successful modeling, they were randomly divided into a model group, a dapagliflozin group (1.5 mg·kg-1·d-1), and high, medium, and low dose groups of Dahuang Tangluo pills (3.6, 1.8, 0.9 g·kg-1·d-1, respectively), with eight mice in each group. All medication groups were administered orally, while the control and model groups were given an equal amount of distilled water by gavage daily. After continuous administration for 10 weeks, the survival status of the mice was observed, and their body weight, FBG, and kidney function-related indicators were measured. Inflammatory indicators in renal tissues were determined by enzyme-linked immunosorbent assay (ELISA). Hematoxylin-eosin (HE) staining, Masson staining, and electron microscopy were used to observe the pathological changes in renal tissues in each group. Immunofluorescence was employed to examine the expression of advanced glycation end products (AGEs) and receptors for advanced glycation end products (RAGE) proteins. Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were utilized to detect the gene and protein expression levels of AGEs, RAGE, inhibitor of nuclear factor-κB (NF-κB) kinase (IKK), and NF-κB in the renal tissues of mice in each group. ResultCompared with control group, the model group showed a significant increase in body weight, FBG, serum creatinine (SCr), urinary microalbumin/urine creatinine ratio (ACR), total cholesterol (TC), and triglycerides (TG) (P<0.05). The levels of intercellular adhesion molecule-1 (ICAM-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) in renal tissues were significantly elevated (P<0.05). Renal histopathological staining and electron microscopy revealed loose arrangement, gaps, structural disarray, mesangial proliferation, and significant fibrosis in renal tissues. Real-time PCR results showed a significant increase in the expression of RAGE, IKK, and NF-κB genes in renal tissues (P<0.05). Immunofluorescence results demonstrated a significant increase in the expression of AGEs and RAGE proteins in renal tissues (P<0.05). Western blot results showed a significant increase in the expression of AGEs, RAGE, IKK, and NF-κB proteins in renal tissues (P<0.05). After drug intervention, compared with model group, the dapagliflozin group and the high-dose Dahuang Tangluo pills group showed significant reductions in body weight, FBG, SCr, and ACR (P<0.05), and a significant decrease in TC in mouse serum (P<0.05), while the high-dose Dahuang Tangluo pills group showed a significant decrease in TG in mouse serum (P<0.05). All treatment groups showed a significant reduction in ICAM-1, IL-6, and TNF-α in renal tissues (P<0.05). Renal histopathological staining and electron microscopy showed improved kidney injury, decreased collagen fiber deposition, and reduced mesangial proliferation in all treatment groups. Real-time PCR results showed a significant decrease in the expression of RAGE, IKK, and NF-κB genes in the dapagliflozin group and the high- and medium-dose Dahuang Tangluo pills groups (P<0.05). Immunofluorescence results demonstrated a significant decrease in the expression of AGEs and RAGE proteins in the dapagliflozin group and the high- and medium-dose Dahuang Tangluo pills groups (P<0.05). Western blot results showed a significant decrease in the expression of AGEs, RAGE, IKK, and NF-κB proteins in the dapagliflozin group and the high- and medium-dose Dahuang Tangluo pills groups (P<0.05). ConclusionDahuang Tangluo pills can improve the pathological structure of the kidneys and reduce renal inflammation in DKD mice, possibly through inhibiting the AGEs/RAGE/IKK/NF-κB pathway.
3.Expression and clinical significance of miR-501-5p and LPAR1 mRNA in esophageal cancer
Yunhui BAI ; Shuangfu YANG ; Kai XU ; Hongmei GAO
Journal of Chinese Physician 2020;22(11):1606-1610
Objective:To study the expression levels of microRNA-501-5p (miR-501-5p) and lysophosphatidic acid receptor 1 (LPAR1) mRNA in esophageal cancer, and to analyze their relationship with clinical pathological data.Methods:From January 2015 to August 2017, 130 patients with esophageal cancer who were hospitalized in Southern Yunnan Central Hospital were selected. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression of miR-501-5p and LPAR1 mRNA in esophageal cancer tissues and adjacent normal tissues; the relationship between miR-501-5p and LPAR1 mRNA and clinical pathological data and the correlation between the two indexes were analyzed. Kaplan Meier test was used to analyze the relation between prognosis and survival of patients.Results:The relative expression of miR-501-5p in esophageal cancer tissues was higher than that in adjacent tissues, and the relative expression of LPAR1 mRNA was lower than in adjacent tissues ( P<0.05). The relative expression of miR-501-5p and LPAR1 mRNA in esophageal cancer tissues were related to tumor tumor node metastasis (TNM) stage, differentiation degree, and lymph node metastasis ( P<0.05), and were not related to patient age, gender, and tumor diameter ( P>0.05). Correlation analysis showed that the expression of miR-501-5p and LPAR1 mRNA in esophageal cancer tissues was negatively correlated ( r=-0.632, P<0.05). Kaplan Meier test showed that the 3-year overall survival rate of patients in high miR-501-5p expression group was lower than that of low expression group, and the 3-year overall survival rate of patients in high LPAR1 mRNA expression group was higher than that of low expression group ( P<0.05). Conclusions:The expression level of miR-501-5p in esophageal cancer is higher than that in adjacent tissues, and the expression level of LPAR1 mRNA is lower than in adjacent tissues. They are related to tumor TNM stage, differentiation degree, and lymph node metastasis and expected to become new biomarkers in the diagnosis and treatment of esophageal cancer.
4.Prostasin gene polymorphism at rs12597511 is associated with severe preeclampsia in Chinese Han women.
Dong LUO ; Yanyan ZHANG ; Yi BAI ; Xijing LIU ; Yunhui GONG ; Bin ZHOU ; Lin ZHANG ; Linli LUO ; Rong ZHOU
Chinese Medical Journal 2014;127(11):2048-2052
BACKGROUNDPreeclampsia, characterized by hypertension and proteinuria, is a multifactorial disease associated with shallow invasion of trophoblast cells and inadequate spiral artery remodeling. Trophoblast and tumor cells have similar invasion mechanism. Prostasin is closely related to tumor development, invasion and metastasis and influences blood pressure through activating epithelial sodium channel. The effect of prostasin on the pathogenesis of preeclampsia remains unclear. This study investigated the association of prostasin gene at rs12597511 with severe preeclampsia.
METHODSA single nucleotide polymorphism, rs12597511, was tested with polymerase chain reaction and restrictionfragment length polymorphism analyses in 179 severe preeclampsia patients and 222 normal pregnant women.
RESULTSThe frequencies of TC + CC genotypes were significantly higher in severe preeclampsia group compared with in control group (the adjusted odds ratio was 2.030, 95% confidence interval 1.195-3.449, P = 0.009). The C allele of rs12597511 was present significantly more often among women with severe preeclampsia (P = 0.001). Genotyping analysis showed that the C allele of rs12597511 could confer a risk for severe preeclampsia.
CONCLUSIONThe higher frequency of C allele of prostasin gene at rs12597511 is associated with severe preeclampsia.
Adult ; Female ; Gene Frequency ; genetics ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Middle Aged ; Polymorphism, Single Nucleotide ; genetics ; Pre-Eclampsia ; genetics ; Pregnancy ; Serine Endopeptidases ; genetics ; Young Adult
5.Construction of 3G intelligent mobile phone-based M-health system model
China Medical Equipment 2014;(7):45-46,47
Objective:Based on the concept of M-health and Characteristics of 3G intelligent mobile phone, constructing a M-health system model of 3G intelligent mobile phone. Methods:The model is based on the communication network, using the 3G intelligent mobile phone, According to the needs of patients, provided by medical personnel whenever and wherever possible. Results: The model completed the basic function of mobile medical system. Conclusion:The paper also discusses the application prospect and existing problems of mobile medical system of 3G intelligent mobile phone.
6.Effect of recipe for kidney tonification on pituitary-thyroid axis and bone density in rat with experimental osteoporosis
Mishan WU ; En LI ; Suzhi ZHAO ; Xia BAI ; Aiying LI ; Yunhui MENG
Chinese Journal of Tissue Engineering Research 2005;9(11):187-189
BACKGROUND: In the research, the recipe for kidney tonification is prepared into "topical form" and "oral form". According to the interrelationship of "acupoints-meridans and collaterals-internal organs-target organs", osteoporosis is treated by the approaches of point compress and oral application respectively. According to the traditional theory of Chinese medicine, it is to infer that the herbs bring the adjustment into play by"being attributive to kidney meridian", but,whether does the specialty of herbs on "targeting medication" happenpractically or not? Hormones in "hypothalamus-pituitary- target gland" system is assayed to further analyze estrogen receptor and androgen receptor of target organs,such as kidney,bone, uterus, thyroid gland and testis so as to observe the relativity between "tropism and receptor".DESIGN: Complete random design with the objects of osteoporosis rats, and the control experimental study with kidney tonification recipe administrate with different approaches.OBJECTIVE: To probe into the effect of kidney tonification recipe on pituitary-thyroid axis in experimental osteoporosis.SETTING: College of Traditional Chinese Medicine of Hebei Medical University.MATERIALS: The experiment was performed in Chinese-Western integrated Basic Experiment Room of Hebei Medical University from January 2000 to December 2004. Sixty SD healthy female rats of 3-month old were employed,weighted(300 ± 20) g. The experimental animals were provided by Hebei Experimental Animal Center. After bred routinely for 1 week in Experimental Room, the rats were randomized into 6 groups,namely normal control(10rats), pathological model group(10 rats), infusing group(10 rats) in which kidney tonification recipe was applied based on 0. 8 g/100 g body mass,acupoint paste on bladder meridian group (10 rats), acupoint paste on kidney meridian group(10 rats) and paste on non-meridian group(10 rats). METHODS: Osteoporosis model was prepared with injection of dexamethasonc. Antagonism osteoporosis acupoint paste(AOAP)on acupoints and non-neridian places was administrated to compare with oral recipe for kidney tonification in the treatment of osteoporosis. It took the changes in serum estrogen, bone density, calcitonin/parathyroid hormone(CT/PTH), thyrotropic hormone(TSH), freeT3andfreeT4asthemeasuringindexes. Immunocyto-chemistry stain and morphological measurement of TSH cell,follicular adenoma of thyroid gland(C cell) and thyroid globulin(Tg) were applied in pituitary gland and thyroid gland to observe the therapeutic results.MAIN OUTCOME MEASURES: ① Adjustment of the recipe for kidney tonification on thyroid function. ② Adjustment of the recipe for kidney tonification on bone density.RESULTS: AOAP improved the levels of estrogen, CT/PTH, TSH, free T3and free T4, increased bone density. The counts of TSH cell, C cell and Tg positive reaction substance were increased and they were colored deeply.CONCLUSION: AOPA, by "multiple relevant convergence reaction", adjusts and affects systematically the endocrinal function of bone metabolism,improves the levels of estrogen and CT/PTH, reverses the declining tendency of pituitary-thyroid axis function and improves thyroid function so as to weaken osteocyte in the bony absorption, enhance osteoblast in osteogenesis and increase bone density.

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