Manual acupuncture relieves microglia-mediated neuroinflammation in a rat model of traumatic brain injury by inhibiting the RhoA/ROCK2 pathway.

OBJECTIVE: To investigate the regulatory mechanism of manual acupuncture (MA) on microglial polarization-mediated neuroinflammation after traumatic brain injury (TBI), focusing on the RhoA/Rho-associated coiled coil-forming protein kinase (ROCK2) pathway. METHODS: Sprague Dawley (SD) rats were used to generate a TBI model using Feeney's freefall epidural impact method. MA was performed on half of the TBI model rats, while the others remained untreated. Acupuncture was administered at GV15, GV16, GV20, GV26, and LI4. At the end of the intervention, rat brain tissue samples were collected, and the microglial M1 polarization status was observed by immunofluorescence labeling of CD86, an M1 microglia-specific protein. RhoA/ROCK2 signaling components were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. An enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of inflammatory factors. RESULTS: Compared with normal rats, the CD86 expression density in the untreated TBI model rats was high and showed an aggregated expression pattern. The genes and proteins of the RhoA/ROCK2 signaling pathway were highly expressed, and inflammatory factors were significantly increased. The CD86 expression density in TBI rats after MA was reduced compared to that in untreated TBI rats and showed a scattered distribution. The expression of RhoA/ROCK2 signaling pathway genes and proteins was also significantly reduced, and inflammatory factors were decreased. CONCLUSION: These results show that MA may inhibit M1 polarization of microglia by regulating the RhoA/ROCK2 signaling pathway, thereby reducing neuroinflammation in TBI.

Compound Kushen Injection as an Adjunctive Therapy for the Treatment of Non-Small-Cell Lung Cancer: A Meta-Analysis of Randomized Controlled Trials.

OBJECTIVES: To evaluate the efficacy and safety of compound Kushen injection (CKI) combined with chemo treatment (chemo) for non-small-cell lung cancer (NSCLC). METHODS: We systematically searched the literature published in seven databases, including Embase, PubMed, central, MEDLINE, CNKI, Wanfang, and VIP, from their inception to April 2019 for all randomized controlled trials (RCTs) comparing CKI plus chemo with chemo alone in patients with NSCLC. Our main end point was clinical efficiency and the secondary outcomes were Karnofsky performance score (KPS), immune function, and adverse events. The Cochrane risk of bias tool was applied for quality assessment. RESULTS: 10 studies involving 1019 participants were included. The clinical response rate (relative risk (RR) = 1.21, 95% confidence interval (CI): 1.06 to 1.37; P=0.003), KPS (RR = 2.18, 95% CI: 1.49 to 3.17; P < 0.0001), immune function (mean differences (MD) = 0.82, 95% CI: 0.12 to 1.52; P=0.02) and adverse effects (RR = 0.67, 95% CI: 0.60 to 0.74; P < 0.00001) in the CKI plus chemo group showed significant differences when compared with chemo alone. CONCLUSIONS: CKI combined with chemo can improve clinical efficiency, KPS, and immune function and reduce adverse reactions in patients with NSCLC when compared with chemo alone. However, more rigorously designed RCTs are needed to validate this benefit, as some of the included RCTs are of low methodological quality.

[Effect of acupuncture intervention on neurological function, cerebral microglia activation and secondary nerve damage in traumatic brain injury rats].

OBJECTIVE: To observe the effect of acupuncture on activities of microglia in traumatic brain injury (TBI) rats. METHODS: Fifty-four male SD rats were randomly and equally divided into normal control, model and acupuncture groups according to the random number table (n＝18 rats in each group). The TBI model was established by using a free fall brain injury striking device after exposing the local cranial bone (to induce the left parietal cerebral contusion). Acupoints "Baihui" (GV20), "Shuigou" (GV26), "Fengfu" (GV16), "Yamen" (GV15) and bilateral "Hegu" (LII4) were stimulated intensively by twirling the filiform needles with force at a range of >360° and a frequency of 160－180 cycles/min for 10 sec in every acupoint, once every 5 min during the 15 minutes' needle retaining. The treatment was given once every day for successive 14 days. The rats of the normal and model groups were grabbed and fixed with the same procedure. The behavioral changes were tested using modified neurological severity score (mNSS). The histopathological changes of the injured cerebral cortex tissues were observed by using hematoxylin-eosin (H.E.) staining, and the fluorescence intensity of Iba-1 (marker of microglia) positive products in the surrounding tissue of the cerebral focus was displayed by immunofluorescence staining, and the contents of neuron specific enolate (NSE) and neurite outgrowth inhibitor-A (Nogo-A) in serum (indicating a secondary nerve damage) were assayed by ELISA. RESULTS: The mNSS scores were significantly increased on day 1, 3, 7 and 14 in the model group in comparison with the normal group (P<0.01) and considerably decreased at the 4 time-points after acupuncture intervention relevant to the model group (P<0.05, P<0.01). H．E. staining showed that modeling induced pathological changes such as the excursion of cell nucleus, cellular swel-ling, vacuole-like change, neuron death, karyopyknosis dissolution, and proliferation of fibrous tissue were relatively milder in the acupuncture group. The average fluorescence intensity values of Iba-1-positive products, serum NSE and Nogo-A contents on day 3, 7 and 14 were significantly higher in the model group than in the normal group (P<0.05, P<0.01), and notably down-regulated in the acupuncture group than in the model group (P<0.05, P<0.01, except Nogo-A on day 3). CONCLUSION: Acupuncture intervention may accelerate neurological function recovery in TBI rats, which is closely related to its effects in inhibiting the activation of microglia and secondary nerve damage.

Effect of acupuncture on the TLR2/4-NF-κB signalling pathway in a rat model of traumatic brain injury.

OBJECTIVE: To study the effect of acupuncture on the TLR2/4-NF-κB signalling pathway in the cortex of Sprague-Dawley rats following traumatic brain injury (TBI), and investigate the possible mechanism underlying the effects of acupuncture on scar repair. METHODS: TBI was established using Feeney's free-falling epidural percussion model. In total, 108 rats were randomly divided into a normal group (n=18), untreated TBI model group (TBI group, n=36) and manual acupuncture-treated TBI group (TBI+MA, n=36). Each group of rats was subdivided into three time groups: 3-day (3d), 7-day (7d) and 14-day (14d). No treatment was given to rats in the normal and TBI groups. The TBI+MA group received manual acupuncture at GV20, GV26, GV16 through GV15, and bilateral LI4. mRNA expression of TLR2, TLR4, NF-κB and protein in the rat cortices was quantified using real-time fluorescence quantitative polymerase chain reaction (qPCR) and Western blot analyses. RESULTS: The modified neurological severity score (mNSS) scores of the TBI+MA group were improved compared with baseline scores 12 hours after modelling, and improved at 7d and 14d compared with the TBI group (P<0.05), while the score of the TBI group did not improve until 14d compared to baseline. mRNA and protein expression of TLR2, TLR4 and NF-κB in the TBI group were higher than the normal group at 3d (P<0.05), reached a peak at 7d, then began to decrease at 14d. mRNA and protein expression of TLR2, TLR4 and NF-κB were higher in the TBI+MA group compared with the TBI group at 3d (P<0.05), were significantly down-regulated at 7d (P<0.01), and decreased to normal levels at 14d. CONCLUSIONS: Acupuncture has a bidirectional regulatory effect on the TLR2/4-NF-κB signalling pathway-related genes TLR2, TLR4 and NF-κB in the TBI rat cortex, promoting their expression in the early stage and inhibiting it in the later stage.