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The meta-analysis revealed that compared to other administration routes of vitamin B12, acupoint injection significantly improved the clinical effectiveness proportion by 28% (risk ratio [RR] 1.28, 95% confidence interval [CI] 1.22–1.35), peroneal nerve motor nerve conduction velocity (MNCV) by 4.43 m/s (MD 4.43, 95% CI 2.83–6.03), peroneal nerve sensory nerve conduction velocity (SNCV) by 3.82 m/s (MD 3.82, 95% CI 3.23–4.41), median nerve MNCV by 5.48 m/s (MD 5.48, 95% CI 4.71–6.24), and median nerve SNCV by 4.62 m/s (MD 4.62, 95% CI 3.84–5.39) in patients with DN, while having no significant impact on fasting blood glucose (FBG) (MD −0.18, 95% CI −0.44 to 0.08), 2-h postprandial blood glucose (2h-PBG) (MD −0.02, 95% CI −0.36 to 0.33), and the adverse event incidence (RR 1.44, 95% CI 0.44–4.70)