Effects of NLRP3 Inflammasome Mediated Pyroptosis on Cardiovascular Diseases and Intervention Mechanism of Chinese Medicine.

Activation of the NOD-like receptor protein 3 (NLRP3) inflammasome signaling pathway is an important mechanism underlying myocardial pyroptosis and plays an important role in inflammatory damage to myocardial tissue in patients with cardiovascular diseases (CVDs), such as diabetic cardiomyopathy, ischemia/reperfusion injury, myocardial infarction, heart failure and hypertension. Noncoding RNAs (ncRNAs) are important regulatory factors. Many Chinese medicine (CM) compounds, including their effective components, can regulate pyroptosis and exert myocardium-protecting effects. The mechanisms underlying this protection include inhibition of inflammasome protein expression, Toll-like receptor 4-NF-κB signal pathway activation, oxidative stress, endoplasmic reticulum stress (ERS), and mixed lineage kinase 3 expression and the regulation of silent information regulator 1. The NLRP3 protein is an important regulatory target for CVD prevention and treatment with CM. Exploring the effects of the interventions mediated by CM and the related mechanisms provides new ideas and perspectives for CVD prevention and treatment.

Transcriptomic analysis of mechanism underlying the effect of induced molting on semen quality and reproductive performance in aged Houdan roosters.

The reproductive performance of breeder roosters has significant economic importance in the poultry industry. Breeder roosters have severely reduced semen quality with age and will be at risk of culling in the following years. In order to extend the use of breeder roosters, we drew on the induced molting model of hens and selected 35 Houdan roosters aged 50 wk for induced molting. By comparing the body weight, testicular weight, semen quality, and reproductive performance before and after induced molting, we found that induced molting could restore the body weight and testicular weight to the levels before molting (P > 0.05). At the same time, it significantly improved sperm motility (P < 0.05) and also improved reproductive performance such as fertilization rate and hatching rate. To further reveal the mechanism underlying the effects of induced molting on semen quality and reproductive performance in aged Houdan roosters, we collected testes from 3 periods: 1 d before fasting (F0), 15 d after fasting (F15), and 32 d after recovery feeding (R32) for transcriptome sequencing analysis. A total of 5,671 genes were detected in F0, F15, and R32, and trend analysis of the 5,671 differential genes showed 2 significant trends (profile 5 and profile 2). KEGG enrichment analysis of the genes in the 2 profiles, revealed significantly enriched pathway regulation of actin cytoskeleton. In the regulation of actin cytoskeleton pathway, we found a protein kinase gene (SRC) and a senescence gene (ROCK2). SRC was highly expressed at F15, leading to the phosphorylation of key substrates, which in turn disrupted the Sertoli cell spermatid connection and the spermiogenesis process, resulting in no mature spermatozoa produced from F15, SRC expression was inhibited at R32, the expression level was reduced, and mature spermatozoa reappeared. The senescence gene ROCK2 was highly expressed at F15 compared to F0 and R32, which may have been responsible for inducing senescence atrophy in the testes.

Corrigendum to "Berberine Reduces Renal Cell Pyroptosis in Golden Hamsters with Diabetic Nephropathy through the Nrf2-NLRP3-Caspase-1-GSDMD Pathway".

[This corrects the article DOI: 10.1155/2021/5545193.].

Mechanisms of Kidney Cell Pyroptosis in Chronic Kidney Disease and the Effects of Traditional Chinese Medicine.

Chronic kidney disease (CKD) is a major public health issue that is highly prevalent worldwide. Pyroptosis is an important pathological mechanism underlying kidney cell damage in CKD and is associated with the classic caspase-1-mediated pathway and nonclassic caspase-4/5/11-mediated pathway. The NLRP3-caspase-1-GSDMD signaling pathway is the key mechanism of kidney cell pyroptosis in CKD, and noncoding RNAs such as lncRNAs and miRNAs are important regulators of kidney cell pyroptosis in CKD. In addition, the NLRP1/AIM2-caspase-1-GSDMD and caspase-3-GSDME signaling pathways have also been shown to mediate kidney cell pyroptosis. Traditional Chinese medicine (TCM) and extracts can interfere with the occurrence and development of kidney cell pyroptosis in CKD by inhibiting the NLRP3 signaling pathway and oxidative stress, activating Nrf-2 signaling, protecting mitochondrial integrity, regulating AMPK signaling, and regulating TXNIP/NLRP3 axis, which have become increasingly prominent. It is critical to explore the effects of TCM on kidney cell pyroptosis in CKD and its mechanisms to identify targets and develop new and effective drugs.

Berberine Reduces Renal Cell Pyroptosis in Golden Hamsters with Diabetic Nephropathy through the Nrf2-NLRP3-Caspase-1-GSDMD Pathway.

Objective: To observe the effect of berberine (BBR) on kidney cell pyroptosis in golden hamsters with diabetic nephropathy (DN) and to explore the molecular mechanism of its renal protection. Methods: Fifty clean-grade male golden hamsters were randomly divided into a control group (10) and a model building group (40). The DN model was established by high-sugar and high-fat feeding and injection of a small amount of STZ. After successful establishment of the model, they were randomly divided into a model group, western medicine group, and berberine high- and low-dose groups. The western medicine group was given irbesartan 13.5 mg/kg, and the berberine high- and low-dose groups were given BBR 200 mg/kg and 100 mg/kg, respectively, for 8 consecutive weeks. An automatic biochemical analyser was used to measure blood glucose, blood lipids, kidney function, MDA, and other indicators; radioimmunoassay was used to assess serum insulin; enzyme-linked immunosorbent assay (ELISA) was used to quantify IL-1ß, IL-6, IL-18, TNF-α; HE, PAS, and Masson staining were used to observe kidney pathological tissue morphology; western blot and real-time fluorescent quantitative PCR were used to assess protein and mRNA expression of molecules, such as Nrf2, NLRP3, Caspase-1, and GSDMD; and TUNEL staining was used to detect DNA damage. SPSS statistical software was used for the data analysis. Results: The kidney tissues of golden hamsters in the control group were normal; Nrf2 was highly expressed, serum MDA level was low, NLRP3 expression in kidney tissue was not obvious, Caspase-1 and GSDMD were weakly expressed, and only a few TUNEL-positive cells were observed. Compared with the control group, the golden hamsters in the model group had obvious renal pathological damage; blood glucose, blood lipids, renal function-related indexes, insulin, and inflammatory factors IL-1ß, IL-6, IL-18, and TNF-α were increased (P < 0.05); NLRP3, Caspase-1, and GSDMD expression was increased; Nrf2 expression was decreased; MDA level was increased (P < 0.05); and the number of TUNEL-positive cells was increased. Compared with the model group, the pathological morphology of the kidney tissue of golden hamsters in the three treatment groups was significantly improved; blood glucose, blood lipids, renal function, and the expression of inflammatory factors IL-1ß and IL-6 were reduced (P < 0.05); NLRP3, Caspase-1, GSDMD, and other molecular proteins and mRNA expression were decreased; Nrf2 expression was increased; MDA level was decreased (P < 0.05); and the number of TUNEL-positive cells was decreased. Conclusion: DN golden hamster kidney NLRP3-Caspase-1-GSDMD signalling was enhanced. BBR can reduce oxidative stress damage by regulating antioxidative Nrf2 and then regulating NLRP3-Caspase-1-GSDMD signalling to inhibit pyroptosis, antagonizing DN inflammation-induced damage.

Low Methoxyl Pectin Protects against Autoimmune Diabetes and Associated Caecal Dysfunction.

SCOPE: This study aims to examine the protective effects of specific low-methoxyl pectin (LMP) on the development of type 1 diabetes (T1D). METHODS AND RESULTS: Female non-obese diabetic (NOD) mice are weaned onto either control or 5% LMP supplemented diets for up to 22 weeks of age. T1D incidence, gut barrier function, and pancreatic-gut immune responses are analyzed. LMP supplementation significantly dampened the onset of T1D in NOD mice. LMP supplementation induces caecal homeostasis, as indicated by the increasing SCFAs production, higher expression of tight junction proteins claudin 1, zonula occludens-2 in caecum. Furthermore, LMP-mediated caecal homeostasis impacts gut-pancreatic immunity, as evidenced by increased regulatory T cell population, modulated inflammatory cytokine expression, and suppressed NOD like receptor protein 3 (NLRP3) inflammasome activation in both caecum and pancreas. CONCLUSION: The data demonstrate that LMP limits T1D development by inducing caecal homeostasis to shape pancreatic immune environment, providing a scientific basis for using LMP as a novel functional supplementation to intervene T1D.

Modulation of Gut Microbiota by Low Methoxyl Pectin Attenuates Type 1 Diabetes in Non-obese Diabetic Mice.

Intestinal homeostasis underpins the development of type 1 diabetes (T1D), and dietary manipulations to enhance intestinal homeostasis have been proposed to prevent T1D. The current study aimed to investigate the efficacy of supplementing a novel specific low-methoxyl pectin (LMP) dietary fiber in preventing T1D development. Female NOD mice were weaned onto control or 5% (wt/wt) LMP supplemented diets for up to 40 weeks of age, overt diabetes incidence and blood glucose were monitored. Then broad-spectrum antibiotics (ABX) treatment per os for 7 days followed by gut microbiota transfer was performed to demonstrate gut microbiota-dependent effects. Next-generation sequencing was used for analyzing the composition of microbiota in caecum. Concentration of short chain fatty acids were determined by GC-MS. The barrier reinforcing tight junction proteins zonula occludens-2 (ZO-2), claudin-1 and NOD like receptor protein 3 (NLRP3) inflammasome activation were determined by Western blot. The proportion of CD25+Foxp3+CD4+ regulatory T cell (Foxp3+ Treg) in the pancreas, pancreatic and mesenteric lymph nodes was analyzed by flow cytometry. We found that LMP supplementation ameliorated T1D development in non-obese diabetic (NOD) mice, as evidenced by decreasing diabetes incidence and fasting glucose levels in LMP fed NOD mice. Further microbiota analysis revealed that LMP supplementation prevented T1D-associated caecal dysbiosis and selectively enriched caecal bacterial species to produce more SCFAs. The LMP-mediated microbial balance further enhanced caecal barrier function and shaped gut-pancreatic immune environment, as characterized by higher expression of tight junction proteins claudin-1, ZO-2 in caecum, increased Foxp3+ Treg population and decreased NLRP3 inflammasome activation in both caecum and pancreas. The microbiota-dependent beneficial effect of LMP on T1D was further proven by the fact that aberration of caecal microbiota by ABX treatment worsened T1D autoimmunity and could be restored with transfer of feces of LMP-fed NOD mice. These data demonstrate that this novel LMP limits T1D development by inducing caecal homeostasis to shape pancreatic immune environment. This finding opens a realistic option for gut microbiota manipulation and prevention of T1D in humans.

[Effect of Huayu Jiedu Recipe on the Expressions of NLRP3, Caspasel, and IL-1 ß in Kidneys of Obstructive Nephropathy Rats].

Objective To observe the effect of Huayu Jiedu Recipe (HJR) on the expressions of nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) , Caspase-1 , IL-1 ß in kidneys of obstructive nephropathy rats. Methods Totally 40 clean grade SD rats were randomly divided into the sham-operation group (n =10) and the model group (n =30). The model of obstructive nephropa- thy was established by unilateral ureteral obstruction (UUO). Totally 30 successfully modeled UUO rats were randomly divided into the model group, the Western medicine group, the Chinese medicine group, 10 in each group. Eplerenone (100 mg . kg ⁻¹ . d⁻¹) was administrated to rats in the Western medicine group. HJR (13.7 g . kg ⁻¹ . d⁻¹) was administrated to rats in the Chinese medicine group. Equal volume of normal saline was administered to rats in the sham-operation group and the model group. All medica- tion was performed once daily for 10 successive days. The serum IL-1 ß level was detected. Protein and mRNA expressions of NLRP3, Caspase-1, and IL-1 ß in renal tissue were detected. TUNEL positive rate was detected by TUNEL method. Results The expression of NLRP3 was not obviously seen, Caspase-1 and IL-1 ß were weakly expressed, and only fewer amount of TUNEL positive cells could be seen in the sham-operation group. Compared with the sham-operation group, serum IL-1ß level increased (P < 0. 01) , mRNA and protein expression of NLRP3, Caspase-1 , and IL-1 ß were up-regulated in renal tissue of the model group (P <0. 01). NLRP3 was mainly expressed in renal interstitial macrophages and renal tubular epithelial cells. Caspase-1 and IL-1 ß were mainly expressed in the cytoplasm of renal tubular epithelial cells. TUNEL positive cells were significantly increased, mainly dominated in interstitial expanded epithelial cells of distal tubules (P <0. 01). Compared with the model group, serum IL-1 ß level was significantly decreased (P <0. 01) ; mRNA and protein expressions of NLRP3, Caspase-1 , and IL- ß were obviously down-regulated (P <0. 01) , and the TUNEL positive rate was obviously decreased (P <0. 05, P < 0. 01) in the two medicated groups. Conclusion HJR could down-regulate mRNA and protein expres- sions of NLRP3, Caspase-1 , and IL-1ß, thus attenuating inflammatory injury of renal tissue.

[Screening of different AFLP fragments between near-isogenic lines of male sterile and fertile Salvia miltiorrhiza and their comparison analysis].

There is distinctive advantage of using male sterile lines to breed new cultivar and produce hybrids, when compared with general breeding method on yield and quality. In our previous work, near-isogenic lines (NILs) of male sterile and fertile Salvia miltiorrhiza have been obtained through continuous hybridization in many years. In this investigation, 378 primer combination were screened by using AFLP and BSA technique, in which 26 markers amplified from seven primers were found to tightly link to male sterile gene. Based on these markers, two linkage genetic maps were constructed. A 2 027,2 028 bp fragment was amplifed from NILs of fertile and sterile S. miltiorrhiza, respectively, using genome walking technique and previous E11/M4-208 marker as template. Four base mutations were found in intron when comparing both fragments. Among all different markers between NILs of male sterile and fertile S. miltiorrhiza, four was found to have 100% identities to chromosome 1, 3 and 5 of Arabidopsis, namely, E01/M09-418, E05/M13-308, E05/M04-750 and E01/M01-204. The E01/M09-418 marker was very close to male sterile gene of S. miltiorrhiza with distance of 2.1 cM, which also had 100% identities to male sterile gene MS2 in Arabidopsis. Both were distributed in chromosome 3 of Arabidopsis. The 2 028 bp fragment also had 100% identities to MS2 gene. Another E05/M04-750 marker that had 100% identities to chromosome 5 of Arabidopsis was found to have high identities to POP085-M05 gene of poplars and low affinity calcium antiporter CAX2 of Arabidopsis with very low E-value. The constructed genetic map and differential fragments with potential functions found in this study provide a solid foundation to lock male sterile genes in S. miltiorrhiza genome and to discover their functions.

[The value of lab diagnostics in Chinese medicine microscopic syndrome differentiation].

Lab diagnostics plays an important role in Chinese medicine (CM) microscopic syndrome differentiation owe to its properties such as microcosmic, objectivity, and quantitation. It is not replaceable in identifying dormant syndrome. Besides, it plays important roles in studying the essence of CM syndromes, the modeling of CM syndromes, the microscopic classification of CM syndromes, and the typing of CM syndromes. Therefore, lab diagnostic indices are important indices for microscopic syndrome differentiation. Microscopic syndrome differentiation deduced from laboratory diagnostics has become beneficial complement for macroscopic syndrome differentiation. It will promote the formation of a brand new syndrome differentiation subject and the development of microscopic syndrome differentiation.

[Lab diagnostics and concepts of integrative medicine].

The integrative medicine concepts were analyzed from a brand-new view angle of lab diagnostics. The microcosmic, objective, early diagnostic, dynamic, and multi-point features of lab diagnostics can be integrated respectively with the macroscopic concept, experience, prevention ahead of disease attack, constant changing concept, and the wholism in Chinese medicine. The concept of integrative medicine in the lab diagnostics is of far-reaching significance in the clinics, research, and teaching of Chinese medicine, as well as the discipline construction of integrative medicine diagnostics. In addition, the complementarity and compatibility of Chinese medicine and lab diagnostics are studied.

[Study on mechanism of the anti-tumor activity of Acanthopanax gracilistylus].

OBJECTIVE: To study the mechanism of anti-tumor activity of Acanthopanax gracilistylus extract (Age). METHODS: The tumor cells proliferation was detected by using (3H)-TdR incorporation method, and the effects of Age on cell cycle of tumor cells, retinoblastoma (Rb) protein and cyclin-dependent kinases (Cdk) were analyzed by flow cytometry and Western blotting assay, respectively. RESULTS: It was indicated by cytoactivity test in vitro that Age only had effect in inhibiting the proliferation of tumor cells, it couldn't lead to death of cells. Under action of Age, the proliferation of tumor cells was halted at G0/G1 stage of cell cycle, and showed no direct cytotoxic effect by Age. Age could induce lowering of the expression of Rb, Cdk2 and Cdk4, cause halt of tumor cell proliferation. CONCLUSION: The tumor inhibitory effect of Age is realized by way of regulating the activity of cell cycle controlling enzymes to suspend the proliferation of tumor cells.

[Experimental study on anti-tumor effects of cortex Acanthopanacis senticosus in vivo and in vitro].

OBJECTIVE: To provide a basis for development and preparation of the new anti-tumor agents from Cortex A-canthopanacis senticosus (CAS), through isolating the active substances from CAS and studying the anti-tumor effect of CAS extracts in vivo and in vitro. METHODS: The effects of CAS extracts and its isolated ingredients on tumor cell proliferation in vitro was determined by 3H-TdR incorporation; the anti-tumor component of CAS was isolated and purified by chromatography; the tumor bearing mice model was established by injecting tumor cell subcutaneously, and the model was used to observe the anti-tumor effect of CAS extract administered through gastrogavage. RESULTS: CAS extract showed obvious inhibition on tumor cell proliferation originated from multiple tissues (P < 0.01) and displayed a better dose-effect relationship. After orally taken CAS extract, the general condition of mice in the experimental group were better than that in the untreated control group, revealing a slower growth and significantly prolonged survival period (P < 0.01). A protein component, having inhibitory effect on tumor cell proliferation and the molecular weight was 64 kda, it was isolated by the thin layer gel chromatography. CONCLUSION: CAS has not only the in vitro inhibitory effect on cell proliferation of multiple kinds of tumor, but also a good anti-tumor effect in vivo. The anti-tumor activity of CAS is correlated with a protein component with the molecular weight of 64 kda. Further isolation, purification, study on mechanism will provide scientific evidence for clinical application and development of CAS in anti-tumor effect.