Mechanism of Yitangkang in Improving Apoptosis of Skeletal Muscle Cells by Inhibiting AGE/RAGE Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo determine the mechanism of Yitangkang in correcting excessive apoptosis of skeletal muscle cells to improve insulin resistance （IR） by inhibiting the advanced glycation end product （AGE）/receptor for the advanced glycation end product （RAGE） signaling pathway. Method① In vitro experiments. Yitangkang-medicated serum was prepared. C2C12 cells were divided into a blank group， a model group， high-， medium-， and low-dose Yitangkang-medicated serum groups （40， 20， and 10 g·kg-1）， and a RAGE inhibitor group. The IR model was induced by palmitic acid in C2C12 cells except for those in the blank group. After the corresponding intervention methods were conducted，the cell viability and glucose consumption level of each group were determined. In addition，the apoptosis rate was determined using flow cytometry. The mRNA and protein expression levels of the important apoptotic proteins ［B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， p53， cysteinyl aspartate-specific protease-3 （Caspase-3）， and cysteinyl aspartate-specific protease-9 （Caspase-9）］ were determined using Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ② In vivo experiments. Ninety-six eligible Wistar rats were divided into a blank group， a model group， high-，medium-，and low-dose Yitangkang groups （40， 20， and 10 g·kg-1）， and a western medicine group （pioglitazone hydrochloride，1.35 mg·kg-1）. The IR model was induced using high-glucose and high-fat feed for diabetes combined with intraperitoneal injection of low-dose streptozotocin （STZ） in animals and verified by the hyperinsulinemic-euglycemic clamp （HEC） test. After the model was determined successfully， the rats in each group were given intragastric administration of drugs as required. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） assay was performed to determine the number of positive apoptotic cells in the skeletal muscle tissues of rats in each group，while Real-time polymerase chain reaction（Real-time PCR） and Western blot were performed to determine the mRNA and protein expression levels of the important apoptotic proteins Bcl-2， Bax， p53， Caspase-3， and Caspase-9. Result① In vitro experiments. compared with the blank group， the model groups showed increased apoptosis rate of C2C12 cells and decreased cell viability and glucose consumption （P<0.01）. Compared with the model group， the Yitangkang-medicated serum groups and the RAGE inhibitor group showed decreased apoptosis rate of C2C12 cells and increased cell viability and glucose consumption （P<0.01）. Compared with the blank group， the model group showed decreased expression levels of Bcl-2 mRNA and protein in C2C12 cells and increased mRNA and protein expression levels of Bax， p53， Caspase-3， and Caspase-9 （P<0.01）. Compared with the model group， the Yitangkang-medicated serum groups and the RAGE inhibitor group showed increased expression levels of Bcl-2 mRNA and protein in C2C12 cells （P<0.01） and decreased mRNA and protein expression levels of Bax， p53， Caspase-3， and Caspase-9 （P<0.05， P<0.01）. ② In vivo experiments. The number of positive apoptotic cells in the skeletal muscle tissues of rats in the model group significantly increased as compared with that in the blank group （P<0.01）. The number of positive apoptotic cells in the skeletal muscle tissues of rats in the Yitangkang groups and the western medicine group decreased as compared with that in the model group （P<0.01）. Compared with the blank group， the model group showed decreased expression levels of Bcl-2 mRNA and protein in skeletal muscle tissues of rats and increased mRNA and protein expression levels of Bax， p53， Caspase-3， and Caspase-9 （P<0.01）. Compared with the model group， the Yitangkang groups and the western medicine group showed increased expression levels of Bcl-2 mRNA and protein in skeletal muscle tissues of rats （P<0.01） and decreased mRNA and protein expression levels of Bax， p53， Caspase-3， and Caspase-9 （P<0.05， P<0.01）. The medium-dose Yitangkang showed a similar effect as RAGE inhibitor， and the effect was equivalent to that of pioglitazone hydrochloride. ConclusionYitangkang can inhibit skeletal muscle cell apoptosis by inhibiting the AGE/RAGE signaling pathway.

Bayesian Network Meta-analysis of Therapeutic Effect of Oral Chinese Patent Medicines on Coronary Artery Disease Complicated with Carotid Atherosclerosis / 中国实验方剂学杂志

ObjectiveTo compare the therapeutic effects of oral Chinese medicines （including Chinese patent medicines） on coronary artery disease （CAD） by the Bayesian network Meta-analysis. MethodThe randomized controlled trials of treating CAD with oral Chinese medicines were retrieved from the China National Knowledge Infrastructure （CNKI）， Wanfang Data， VIP， PubMed， Web of Science， Embase， and Cochrane Library from the inception to December 1， 2022. The Cochrane risk of bias assessment tool was used to evaluate the quality of the included articles. The direct meta-analysis was performed to compare the performance of oral Chinese medicines alone and in combination with Western medicine in the treatment of CAD in terms of intima-media thickness （IMT）， vascular endothelial function， plaque score， hypersensitive C-reactive protein （hs-CRP）， total cholesterol （TC）， triglyceride （TG）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， and total response rate. Furthermore， the Bayesian network Meta-analysis was performed to compare the therapeutic effects of different Chinese medicines. ResultA total of 41 articles were included. The direct meta-analysis results showed that Chinese medicines combined with Western medicine outperformed Western medicine alone in recovering all the indicators of CAD. The Bayesian network meta-analysis yielded the following results. In terms of the total response rate， modified Huangqi Guizhi Wuwutang and Sanqi Huayu pills had obvious advantages over other Chinese medicines. In terms of IMT and plaque score， Xiaoban Huazhuo decoction， Yiqi Tongluo formula， Ruangan Jiangzhi capsules， and Guanxin Shutong capsules had obvious advantages over other Chinese medicines. In terms of blood lipid indicators， Shenqi Roumai mixture， Ruangan Jiangzhi capsules， Xiaoban Huazhuo decoction， Qiwei Sanxiong decoction， and Sanqi Huayu pills were superior to other Chinese medicines. The Chinese medicines above mainly had the functions of activating blood， resolving stasis， resolving phlegm， and dredging vessels. ConclusionThe combination of oral Chinese medicines and Western medicine is effective in treating CAD. Clinicians can use the drugs targeting abnormal indicators according to the results of this Bayesian network meta-analysis combined with the actual situation of patients to achieve better therapeutic effects.

Protein and RNA expression of acid-sensing ion channels 2 and 3 in myocardial ischemia rats induced by isoproterenol.

OBJECTIVE: To investigate the impact of electro-acupuncture at the Neiguan (PC 6) acupoint on protein and RNA expression of acid-sensing ion channel 2 (ASIC2) and ASIC3 in myocardial ischemia rats. METHODS: Fifty male Sprague-Dawley rats were used, weighing (230 ± 50) g. The rats were randomized into a normal group A, model group B, Neiguan (PC 6) group C, Lieque (LU 7) group D, and A-shi points group E. There were 10 rats in each group. Rats were continuously administered 85 mg/ kg intravenous isoproterenol daily to establish the model. Successfully modeled rats in groups C, D, and E were given electro-acupuncture treatment. Each group of rats was sacrificed with chloral hydrate (1 mL/100 g) intraperitoneal injection. The left ventricular myocardium was extracted and placed at -70 °C until use. Western blot analysis and real-time PCR were performed to assay protein and RNA expressions of ASIC2 and ASIC3, respectively. Fold changes in RNA expression were quantified with the 2^ (-ΔΔCt) method. Blood samples were drawn from the aorta abdominalis and tested for creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) levels using enzyme-linked immunosorbent assay. RESULTS: Myocardial ischemia rats given electro-acupuncture at the Neiguan (PC 6) acupoint had significantly lower protein and RNA expression of ASIC2 and ASIC3, and CK-MB and LDH levels, compared with model rats (P < 0.01). CONCLUSION: Electro-acupuncture at the Neiguan (PC 6) acupoint can not only decrease the protein and RNA expression of ASIC2 and ASIC3, but also inhibit the opening of ASICs and reduce the cardiomyocyte damage in myocardial ischemia rats.

The effect of Chinese herbal compound Yitangkang extract on expression of renal AGEs and RAGEs in rats with diabetic nephropathy / 中国生化药物杂志

Objective To observe the effect of Chinese herbal compound Yitangkang on the expressions of advanced glyclation end-products (AGEs) and the receptor of advanced glyclation end-products mRNA(RAGE-mRNA) in rats with diabetic nephropathy(DN),and to discuss the mechanism of the formulas in the treatment of DN. Methods The rat model of DN were established by intraperitoneal injection of strepotozotocin(STZ). After 8 weeks intervention with medications, the expression of renal RAGEs mRNA was detected by Real-time PCR, renal RAGE by immunohistochemistry, and renal content of AGEs by spectrophotometry. Results Compared with normal control group, the expression of renal RAGEs mRNA and RAGEs in diabetic nephropathy rats were increased significantly(P<0.01). Compared with DN group, the expression of renal RAGEs mRNA and RAGEs in three doses Yitangkang groups and western medicine group were decreased significantly(P<0.01). Conclusion Chinese herbal compound Yitangkang could protect kidney of diabetic nephropathy rats,and the mechanism is related to its loweirng renal expression of RAGEs mRNA, RAGEs and AGEs content.