1.Optimized CRISPR/Cas9 System in Brain Science and Application Prospects in Field of Traditional Chinese Medicine
Shuoqiu DENG ; Shuiqing QU ; Yu ZHANG ; Yuanmin YANG ; Zhongyuan ZHENG ; Tuo LIU ; Lina CHEN ; Yujie LI
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(1):169-180
Clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR associated nuclease 9 (CRISPR/Cas9) is a self-defense system found in bacteria and archaea that enables targeted gene editing based on the principle. Due to its universality, efficiency, and simplicity, CRISPR/Cas9 has been applied in the pathological mechanism and prevention and treatment of diseases in many fields. Cerebrovascular diseases and central nervous system diseases seriously endanger human health. Stroke is related to genetics, unhealthy living habits, chronic diseases, and other factors. The brain tissue structure is complex and the cell types are diverse. It is difficult for a universal gene editing platform to study target genes safely, specifically, and efficiently. Scholars have continuously improved and optimized gene editing technology, explored the potential and research methods of gene editing technology, and promoted the research process of brain science. After a brief introduction to the mechanism of CRISPR/Cas9, this paper mainly summarized the optimization of the system in the fields of cerebral science including delivery methods, adeno-associated virus assembly, and new nanomaterials. Its application in cerebrovascular research including vascular homeostasis, microglial homeostasis, angiogenesis, blood-brain barrier, and drug screening was also summarized. Finally, this paper prospected the development of CRISPR/Cas9 in traditional Chinese medicine, hoping to provide references for related research design.
2.Protective Effect of Shenlian Prescription on Acute Lung Injury Induced by Particulate Matter Exposure in Rats
Yuanmin YANG ; Shuiqing QU ; Lina CHEN ; Shuoqiu DENG ; Yu ZHANG ; Zhongyuan ZHENG ; Xiaoxin ZHU ; Yuxiang LI ; Yujie LI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(20):37-44
ObjectiveTo observe the protective effect of Shenlian prescription on acute lung injury induced by particulate matter (PM) exposure in rats and explore the mechanism. MethodFifty male SD rats were randomly divided into the control group, model group, Shenlian low-dose group (4.32 g·kg-1), Shenlian high-dose group (8.64 g·kg-1), and roflumilast group (3.46 mg·kg-1), with 10 in each group. Pre-administration with drugs by gavage was performed for one week. On the 8th and 11th days, the control group was instilled with normal saline in the trachea and the other groups with PM suspension to establish a rat model of acute lung injury induced by PM exposure. After modeling, drugs were given continuously until the end of the experiment. Forty-eight hours after the last exposure, the lung function of rats was detected. Then the rats were sacrificed and the lung morphological changes and pathological changes by hematoxylin-eosin (HE) staining were observed. CD68 expression in lung was detected by immunohistochemistry, and the levels of lung injury markers surfactant protein A (SP-A) and Clara cell protein16 (CC16) in serum were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of interleukin-1α (IL-1α), IL-6, IL-18, and monocyte chemoattractant protein-1 (MCP-1) in lung tissue was measured by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultCompared with those in the control group, the rats in the model group had decreased lung function and obvious structural damage of lung tissue, PM deposition, and infiltration of CD68 positive cells. The expressions of IL-1α, IL-6, IL-18, and MCP-1 in lung tissue were increased (P<0.01). Compared with the model group, Shenlian prescription low and high doses restored the rats' lung function injury(P<0.05,P<0.01), improved lung morphological and pathological structure, and reduced PM deposition. Infiltration of CD68 positive cells in lung was not significantly decreased. The levels of inflammatory factors IL-1α, IL -6, IL-18, and MCP-1 in lung were lowered (P<0.01). ConclusionShenlian prescription could protect the rats' lung injury caused by PM exposure, improve lung morphology, and reduce PM deposition and inflammatory factor expression.